WO2017074159A2 - Coumestrol-containing germinated fermented soybean extract having hepatocyte protective effect and bone density increase effect and composition containing same - Google Patents
Coumestrol-containing germinated fermented soybean extract having hepatocyte protective effect and bone density increase effect and composition containing same Download PDFInfo
- Publication number
- WO2017074159A2 WO2017074159A2 PCT/KR2016/012399 KR2016012399W WO2017074159A2 WO 2017074159 A2 WO2017074159 A2 WO 2017074159A2 KR 2016012399 W KR2016012399 W KR 2016012399W WO 2017074159 A2 WO2017074159 A2 WO 2017074159A2
- Authority
- WO
- WIPO (PCT)
- Prior art keywords
- fermented soybean
- germinated fermented
- soybean extract
- germinated
- extract
- Prior art date
Links
- 235000020712 soy bean extract Nutrition 0.000 title claims abstract description 44
- 239000000203 mixture Substances 0.000 title claims abstract description 21
- 230000037182 bone density Effects 0.000 title claims abstract description 10
- 210000003494 hepatocyte Anatomy 0.000 title claims abstract description 6
- 230000000694 effects Effects 0.000 title abstract description 27
- ZZIALNLLNHEQPJ-UHFFFAOYSA-N coumestrol Chemical compound C1=C(O)C=CC2=C1OC(=O)C1=C2OC2=CC(O)=CC=C12 ZZIALNLLNHEQPJ-UHFFFAOYSA-N 0.000 title abstract description 8
- 230000001681 protective effect Effects 0.000 title description 5
- 230000001965 increasing effect Effects 0.000 claims abstract description 39
- 235000010469 Glycine max Nutrition 0.000 claims description 68
- 244000068988 Glycine max Species 0.000 claims description 67
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 19
- 238000000034 method Methods 0.000 claims description 13
- 239000000284 extract Substances 0.000 claims description 10
- 235000013305 food Nutrition 0.000 claims description 10
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 9
- 239000000546 pharmaceutical excipient Substances 0.000 claims description 6
- 239000008187 granular material Substances 0.000 claims description 5
- 239000002775 capsule Substances 0.000 claims description 4
- 238000009472 formulation Methods 0.000 claims description 4
- 230000000144 pharmacologic effect Effects 0.000 claims description 3
- 238000001914 filtration Methods 0.000 claims description 2
- 239000007788 liquid Substances 0.000 claims description 2
- 239000006187 pill Substances 0.000 claims description 2
- 238000001035 drying Methods 0.000 claims 2
- 239000002417 nutraceutical Substances 0.000 claims 2
- 235000021436 nutraceutical agent Nutrition 0.000 claims 2
- 241000894006 Bacteria Species 0.000 claims 1
- 241000219061 Rheum Species 0.000 claims 1
- 239000008194 pharmaceutical composition Substances 0.000 claims 1
- 238000002525 ultrasonication Methods 0.000 claims 1
- 210000000988 bone and bone Anatomy 0.000 abstract description 31
- 238000005259 measurement Methods 0.000 abstract description 29
- 230000003078 antioxidant effect Effects 0.000 abstract description 16
- 229910052500 inorganic mineral Inorganic materials 0.000 abstract description 15
- 239000011707 mineral Substances 0.000 abstract description 15
- 238000010171 animal model Methods 0.000 abstract description 14
- 230000009245 menopause Effects 0.000 abstract description 10
- 238000012360 testing method Methods 0.000 abstract description 10
- 208000024891 symptom Diseases 0.000 abstract description 9
- 238000000338 in vitro Methods 0.000 abstract description 8
- 231100000304 hepatotoxicity Toxicity 0.000 abstract description 7
- 206010019851 Hepatotoxicity Diseases 0.000 abstract description 4
- 230000003833 cell viability Effects 0.000 abstract description 4
- 230000002489 hematologic effect Effects 0.000 abstract description 4
- 230000007686 hepatotoxicity Effects 0.000 abstract description 4
- 238000002360 preparation method Methods 0.000 abstract description 3
- 230000002611 ovarian Effects 0.000 description 34
- 239000000262 estrogen Substances 0.000 description 33
- 229940011871 estrogen Drugs 0.000 description 33
- 210000004369 blood Anatomy 0.000 description 22
- 239000008280 blood Substances 0.000 description 22
- 210000004027 cell Anatomy 0.000 description 22
- RWSXRVCMGQZWBV-WDSKDSINSA-N glutathione Chemical compound OC(=O)[C@@H](N)CCC(=O)N[C@@H](CS)C(=O)NCC(O)=O RWSXRVCMGQZWBV-WDSKDSINSA-N 0.000 description 19
- MUMGGOZAMZWBJJ-DYKIIFRCSA-N Testostosterone Chemical compound O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 MUMGGOZAMZWBJJ-DYKIIFRCSA-N 0.000 description 16
- 238000002271 resection Methods 0.000 description 15
- 206010027304 Menopausal symptoms Diseases 0.000 description 14
- 235000010755 mineral Nutrition 0.000 description 14
- 102000004190 Enzymes Human genes 0.000 description 12
- 108090000790 Enzymes Proteins 0.000 description 12
- 210000004185 liver Anatomy 0.000 description 12
- 102000002260 Alkaline Phosphatase Human genes 0.000 description 11
- 108020004774 Alkaline Phosphatase Proteins 0.000 description 11
- 102000019197 Superoxide Dismutase Human genes 0.000 description 11
- 108010012715 Superoxide dismutase Proteins 0.000 description 11
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 description 11
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 description 10
- 102000005720 Glutathione transferase Human genes 0.000 description 10
- 108010070675 Glutathione transferase Proteins 0.000 description 10
- 230000007423 decrease Effects 0.000 description 9
- 239000008103 glucose Substances 0.000 description 9
- 229960003180 glutathione Drugs 0.000 description 9
- 230000003247 decreasing effect Effects 0.000 description 8
- 230000035784 germination Effects 0.000 description 8
- 229960003604 testosterone Drugs 0.000 description 8
- 208000001132 Osteoporosis Diseases 0.000 description 7
- 238000002679 ablation Methods 0.000 description 7
- 238000002474 experimental method Methods 0.000 description 7
- 150000002632 lipids Chemical class 0.000 description 7
- 210000002966 serum Anatomy 0.000 description 7
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 description 6
- 241001465754 Metazoa Species 0.000 description 6
- OHDRQQURAXLVGJ-HLVWOLMTSA-N azane;(2e)-3-ethyl-2-[(e)-(3-ethyl-6-sulfo-1,3-benzothiazol-2-ylidene)hydrazinylidene]-1,3-benzothiazole-6-sulfonic acid Chemical compound [NH4+].[NH4+].S/1C2=CC(S([O-])(=O)=O)=CC=C2N(CC)C\1=N/N=C1/SC2=CC(S([O-])(=O)=O)=CC=C2N1CC OHDRQQURAXLVGJ-HLVWOLMTSA-N 0.000 description 6
- 238000010603 microCT Methods 0.000 description 6
- 150000008442 polyphenolic compounds Chemical class 0.000 description 6
- 235000013824 polyphenols Nutrition 0.000 description 6
- 239000003642 reactive oxygen metabolite Substances 0.000 description 6
- 235000019786 weight gain Nutrition 0.000 description 6
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 5
- 208000017657 Menopausal disease Diseases 0.000 description 5
- 208000008589 Obesity Diseases 0.000 description 5
- 235000010627 Phaseolus vulgaris Nutrition 0.000 description 5
- 244000046052 Phaseolus vulgaris Species 0.000 description 5
- 239000011575 calcium Substances 0.000 description 5
- 229910052791 calcium Inorganic materials 0.000 description 5
- 235000012000 cholesterol Nutrition 0.000 description 5
- 230000036541 health Effects 0.000 description 5
- 229940088597 hormone Drugs 0.000 description 5
- 239000005556 hormone Substances 0.000 description 5
- CJWQYWQDLBZGPD-UHFFFAOYSA-N isoflavone Natural products C1=C(OC)C(OC)=CC(OC)=C1C1=COC2=C(C=CC(C)(C)O3)C3=C(OC)C=C2C1=O CJWQYWQDLBZGPD-UHFFFAOYSA-N 0.000 description 5
- 150000002515 isoflavone derivatives Chemical class 0.000 description 5
- 235000008696 isoflavones Nutrition 0.000 description 5
- 239000000463 material Substances 0.000 description 5
- 235000020824 obesity Nutrition 0.000 description 5
- 150000003839 salts Chemical class 0.000 description 5
- 230000028327 secretion Effects 0.000 description 5
- 239000000126 substance Substances 0.000 description 5
- 238000011282 treatment Methods 0.000 description 5
- 230000004584 weight gain Effects 0.000 description 5
- VFNKZQNIXUFLBC-UHFFFAOYSA-N 2',7'-dichlorofluorescein Chemical compound O1C(=O)C2=CC=CC=C2C21C1=CC(Cl)=C(O)C=C1OC1=C2C=C(Cl)C(O)=C1 VFNKZQNIXUFLBC-UHFFFAOYSA-N 0.000 description 4
- OKKJLVBELUTLKV-UHFFFAOYSA-N Methanol Chemical compound OC OKKJLVBELUTLKV-UHFFFAOYSA-N 0.000 description 4
- OAICVXFJPJFONN-UHFFFAOYSA-N Phosphorus Chemical compound [P] OAICVXFJPJFONN-UHFFFAOYSA-N 0.000 description 4
- 241000700159 Rattus Species 0.000 description 4
- WQZGKKKJIJFFOK-VFUOTHLCSA-N beta-D-glucose Chemical compound OC[C@H]1O[C@@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-VFUOTHLCSA-N 0.000 description 4
- 230000004097 bone metabolism Effects 0.000 description 4
- 230000008859 change Effects 0.000 description 4
- 239000000469 ethanolic extract Substances 0.000 description 4
- 238000000855 fermentation Methods 0.000 description 4
- 230000004151 fermentation Effects 0.000 description 4
- 238000001727 in vivo Methods 0.000 description 4
- 230000004060 metabolic process Effects 0.000 description 4
- 210000003470 mitochondria Anatomy 0.000 description 4
- 239000011574 phosphorus Substances 0.000 description 4
- 229910052698 phosphorus Inorganic materials 0.000 description 4
- 238000000926 separation method Methods 0.000 description 4
- VOXZDWNPVJITMN-ZBRFXRBCSA-N 17β-estradiol Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 VOXZDWNPVJITMN-ZBRFXRBCSA-N 0.000 description 3
- 102000016938 Catalase Human genes 0.000 description 3
- 108010053835 Catalase Proteins 0.000 description 3
- 241000196324 Embryophyta Species 0.000 description 3
- 108010023302 HDL Cholesterol Proteins 0.000 description 3
- 108010028554 LDL Cholesterol Proteins 0.000 description 3
- 238000002835 absorbance Methods 0.000 description 3
- 230000032683 aging Effects 0.000 description 3
- 238000004458 analytical method Methods 0.000 description 3
- 239000003963 antioxidant agent Substances 0.000 description 3
- 230000037396 body weight Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 239000012153 distilled water Substances 0.000 description 3
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 3
- 229960005309 estradiol Drugs 0.000 description 3
- 229930182833 estradiol Natural products 0.000 description 3
- 235000013402 health food Nutrition 0.000 description 3
- 208000006454 hepatitis Diseases 0.000 description 3
- 230000006872 improvement Effects 0.000 description 3
- 210000005229 liver cell Anatomy 0.000 description 3
- 230000007056 liver toxicity Effects 0.000 description 3
- 238000004519 manufacturing process Methods 0.000 description 3
- 230000002438 mitochondrial effect Effects 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 210000003205 muscle Anatomy 0.000 description 3
- 239000013642 negative control Substances 0.000 description 3
- 210000001672 ovary Anatomy 0.000 description 3
- 230000036542 oxidative stress Effects 0.000 description 3
- 239000003075 phytoestrogen Substances 0.000 description 3
- 239000000843 powder Substances 0.000 description 3
- 230000003449 preventive effect Effects 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 229940088594 vitamin Drugs 0.000 description 3
- 229930003231 vitamin Natural products 0.000 description 3
- 235000013343 vitamin Nutrition 0.000 description 3
- 239000011782 vitamin Substances 0.000 description 3
- YBJHBAHKTGYVGT-ZKWXMUAHSA-N (+)-Biotin Chemical compound N1C(=O)N[C@@H]2[C@H](CCCCC(=O)O)SC[C@@H]21 YBJHBAHKTGYVGT-ZKWXMUAHSA-N 0.000 description 2
- GVJHHUAWPYXKBD-UHFFFAOYSA-N (±)-α-Tocopherol Chemical compound OC1=C(C)C(C)=C2OC(CCCC(C)CCCC(C)CCCC(C)C)(C)CCC2=C1C GVJHHUAWPYXKBD-UHFFFAOYSA-N 0.000 description 2
- 208000000044 Amnesia Diseases 0.000 description 2
- 208000019901 Anxiety disease Diseases 0.000 description 2
- 206010065687 Bone loss Diseases 0.000 description 2
- 208000024172 Cardiovascular disease Diseases 0.000 description 2
- 229920002261 Corn starch Polymers 0.000 description 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 208000033830 Hot Flashes Diseases 0.000 description 2
- 206010060800 Hot flush Diseases 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 102000007330 LDL Lipoproteins Human genes 0.000 description 2
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 description 2
- 206010067125 Liver injury Diseases 0.000 description 2
- TWRXJAOTZQYOKJ-UHFFFAOYSA-L Magnesium chloride Chemical compound [Mg+2].[Cl-].[Cl-] TWRXJAOTZQYOKJ-UHFFFAOYSA-L 0.000 description 2
- 208000026139 Memory disease Diseases 0.000 description 2
- 206010029216 Nervousness Diseases 0.000 description 2
- DFPAKSUCGFBDDF-UHFFFAOYSA-N Nicotinamide Chemical compound NC(=O)C1=CC=CN=C1 DFPAKSUCGFBDDF-UHFFFAOYSA-N 0.000 description 2
- 108010073771 Soybean Proteins Proteins 0.000 description 2
- 235000010724 Wisteria floribunda Nutrition 0.000 description 2
- XLOMVQKBTHCTTD-UHFFFAOYSA-N Zinc monoxide Chemical compound [Zn]=O XLOMVQKBTHCTTD-UHFFFAOYSA-N 0.000 description 2
- 239000004480 active ingredient Substances 0.000 description 2
- 231100000354 acute hepatitis Toxicity 0.000 description 2
- 235000006708 antioxidants Nutrition 0.000 description 2
- 230000036506 anxiety Effects 0.000 description 2
- 235000010323 ascorbic acid Nutrition 0.000 description 2
- 239000011668 ascorbic acid Substances 0.000 description 2
- 229960005070 ascorbic acid Drugs 0.000 description 2
- QVGXLLKOCUKJST-UHFFFAOYSA-N atomic oxygen Chemical compound [O] QVGXLLKOCUKJST-UHFFFAOYSA-N 0.000 description 2
- 235000013361 beverage Nutrition 0.000 description 2
- 210000000481 breast Anatomy 0.000 description 2
- 239000000969 carrier Substances 0.000 description 2
- 239000008120 corn starch Substances 0.000 description 2
- 210000000172 cytosol Anatomy 0.000 description 2
- 230000006378 damage Effects 0.000 description 2
- 230000001419 dependent effect Effects 0.000 description 2
- 238000001784 detoxification Methods 0.000 description 2
- 235000018823 dietary intake Nutrition 0.000 description 2
- 230000001076 estrogenic effect Effects 0.000 description 2
- 239000004744 fabric Substances 0.000 description 2
- OVBPIULPVIDEAO-LBPRGKRZSA-N folic acid Chemical compound C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)N[C@@H](CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-LBPRGKRZSA-N 0.000 description 2
- 230000037406 food intake Effects 0.000 description 2
- 229930195712 glutamate Natural products 0.000 description 2
- 231100000234 hepatic damage Toxicity 0.000 description 2
- 238000002657 hormone replacement therapy Methods 0.000 description 2
- 238000003384 imaging method Methods 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 229940041476 lactose 100 mg Drugs 0.000 description 2
- 230000003859 lipid peroxidation Effects 0.000 description 2
- 230000008818 liver damage Effects 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 230000006984 memory degeneration Effects 0.000 description 2
- 208000023060 memory loss Diseases 0.000 description 2
- VMGAPWLDMVPYIA-HIDZBRGKSA-N n'-amino-n-iminomethanimidamide Chemical compound N\N=C\N=N VMGAPWLDMVPYIA-HIDZBRGKSA-N 0.000 description 2
- 229910052760 oxygen Inorganic materials 0.000 description 2
- 239000001301 oxygen Substances 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- BOLDJAUMGUJJKM-LSDHHAIUSA-N renifolin D Natural products CC(=C)[C@@H]1Cc2c(O)c(O)ccc2[C@H]1CC(=O)c3ccc(O)cc3O BOLDJAUMGUJJKM-LSDHHAIUSA-N 0.000 description 2
- 229940001941 soy protein Drugs 0.000 description 2
- 235000000346 sugar Nutrition 0.000 description 2
- 238000001356 surgical procedure Methods 0.000 description 2
- 239000003826 tablet Substances 0.000 description 2
- 210000001519 tissue Anatomy 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 230000001988 toxicity Effects 0.000 description 2
- 231100000419 toxicity Toxicity 0.000 description 2
- 150000003626 triacylglycerols Chemical class 0.000 description 2
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 2
- 210000000689 upper leg Anatomy 0.000 description 2
- 150000003722 vitamin derivatives Chemical class 0.000 description 2
- FPIPGXGPPPQFEQ-UHFFFAOYSA-N 13-cis retinol Natural products OCC=C(C)C=CC=C(C)C=CC1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-UHFFFAOYSA-N 0.000 description 1
- TWCMVXMQHSVIOJ-UHFFFAOYSA-N Aglycone of yadanzioside D Natural products COC(=O)C12OCC34C(CC5C(=CC(O)C(O)C5(C)C3C(O)C1O)C)OC(=O)C(OC(=O)C)C24 TWCMVXMQHSVIOJ-UHFFFAOYSA-N 0.000 description 1
- 206010002091 Anaesthesia Diseases 0.000 description 1
- 244000105624 Arachis hypogaea Species 0.000 description 1
- 241000228212 Aspergillus Species 0.000 description 1
- 240000006439 Aspergillus oryzae Species 0.000 description 1
- 235000002247 Aspergillus oryzae Nutrition 0.000 description 1
- PLMKQQMDOMTZGG-UHFFFAOYSA-N Astrantiagenin E-methylester Natural products CC12CCC(O)C(C)(CO)C1CCC1(C)C2CC=C2C3CC(C)(C)CCC3(C(=O)OC)CCC21C PLMKQQMDOMTZGG-UHFFFAOYSA-N 0.000 description 1
- WDPFQABQVGJEBZ-MAKOZQESSA-N Bothermon Chemical compound OC1=CC=C2[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1.O=C1CC[C@]2(C)[C@H]3CC[C@](C)([C@H](CC4)O)[C@@H]4[C@@H]3CCC2=C1 WDPFQABQVGJEBZ-MAKOZQESSA-N 0.000 description 1
- 206010006187 Breast cancer Diseases 0.000 description 1
- 208000026310 Breast neoplasm Diseases 0.000 description 1
- BVKZGUZCCUSVTD-UHFFFAOYSA-L Carbonate Chemical compound [O-]C([O-])=O BVKZGUZCCUSVTD-UHFFFAOYSA-L 0.000 description 1
- 206010008909 Chronic Hepatitis Diseases 0.000 description 1
- AUNGANRZJHBGPY-UHFFFAOYSA-N D-Lyxoflavin Natural products OCC(O)C(O)C(O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-UHFFFAOYSA-N 0.000 description 1
- ZZZCUOFIHGPKAK-UHFFFAOYSA-N D-erythro-ascorbic acid Natural products OCC1OC(=O)C(O)=C1O ZZZCUOFIHGPKAK-UHFFFAOYSA-N 0.000 description 1
- 101710088194 Dehydrogenase Proteins 0.000 description 1
- ZNZYKNKBJPZETN-WELNAUFTSA-N Dialdehyde 11678 Chemical compound N1C2=CC=CC=C2C2=C1[C@H](C[C@H](/C(=C/O)C(=O)OC)[C@@H](C=C)C=O)NCC2 ZNZYKNKBJPZETN-WELNAUFTSA-N 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 229930091371 Fructose Natural products 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 239000005715 Fructose Substances 0.000 description 1
- 108010024636 Glutathione Proteins 0.000 description 1
- 102000005744 Glycoside Hydrolases Human genes 0.000 description 1
- 108010031186 Glycoside Hydrolases Proteins 0.000 description 1
- 102000015779 HDL Lipoproteins Human genes 0.000 description 1
- 206010019663 Hepatic failure Diseases 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 208000035150 Hypercholesterolemia Diseases 0.000 description 1
- 108010044467 Isoenzymes Proteins 0.000 description 1
- PIWKPBJCKXDKJR-UHFFFAOYSA-N Isoflurane Chemical compound FC(F)OC(Cl)C(F)(F)F PIWKPBJCKXDKJR-UHFFFAOYSA-N 0.000 description 1
- 108010007622 LDL Lipoproteins Proteins 0.000 description 1
- 231100000002 MTT assay Toxicity 0.000 description 1
- 238000000134 MTT assay Methods 0.000 description 1
- OVBPIULPVIDEAO-UHFFFAOYSA-N N-Pteroyl-L-glutaminsaeure Natural products C=1N=C2NC(N)=NC(=O)C2=NC=1CNC1=CC=C(C(=O)NC(CCC(O)=O)C(O)=O)C=C1 OVBPIULPVIDEAO-UHFFFAOYSA-N 0.000 description 1
- SEBFKMXJBCUCAI-UHFFFAOYSA-N NSC 227190 Natural products C1=C(O)C(OC)=CC(C2C(OC3=CC=C(C=C3O2)C2C(C(=O)C3=C(O)C=C(O)C=C3O2)O)CO)=C1 SEBFKMXJBCUCAI-UHFFFAOYSA-N 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 206010031149 Osteitis Diseases 0.000 description 1
- 102000004160 Phosphoric Monoester Hydrolases Human genes 0.000 description 1
- 108090000608 Phosphoric Monoester Hydrolases Proteins 0.000 description 1
- LCTONWCANYUPML-UHFFFAOYSA-M Pyruvate Chemical compound CC(=O)C([O-])=O LCTONWCANYUPML-UHFFFAOYSA-M 0.000 description 1
- AUNGANRZJHBGPY-SCRDCRAPSA-N Riboflavin Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)CN1C=2C=C(C)C(C)=CC=2N=C2C1=NC(=O)NC2=O AUNGANRZJHBGPY-SCRDCRAPSA-N 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- OUUQCZGPVNCOIJ-UHFFFAOYSA-M Superoxide Chemical compound [O-][O] OUUQCZGPVNCOIJ-UHFFFAOYSA-M 0.000 description 1
- 238000008050 Total Bilirubin Reagent Methods 0.000 description 1
- 102000003929 Transaminases Human genes 0.000 description 1
- 108090000340 Transaminases Proteins 0.000 description 1
- 208000002495 Uterine Neoplasms Diseases 0.000 description 1
- FPIPGXGPPPQFEQ-BOOMUCAASA-N Vitamin A Natural products OC/C=C(/C)\C=C\C=C(\C)/C=C/C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-BOOMUCAASA-N 0.000 description 1
- 229930003779 Vitamin B12 Natural products 0.000 description 1
- 229930003471 Vitamin B2 Natural products 0.000 description 1
- 229930003268 Vitamin C Natural products 0.000 description 1
- 229930003427 Vitamin E Natural products 0.000 description 1
- TVXBFESIOXBWNM-UHFFFAOYSA-N Xylitol Natural products OCCC(O)C(O)C(O)CCO TVXBFESIOXBWNM-UHFFFAOYSA-N 0.000 description 1
- 230000009471 action Effects 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 210000001789 adipocyte Anatomy 0.000 description 1
- FPIPGXGPPPQFEQ-OVSJKPMPSA-N all-trans-retinol Chemical compound OC\C=C(/C)\C=C\C=C(/C)\C=C\C1=C(C)CCCC1(C)C FPIPGXGPPPQFEQ-OVSJKPMPSA-N 0.000 description 1
- XAGFODPZIPBFFR-UHFFFAOYSA-N aluminium Chemical compound [Al] XAGFODPZIPBFFR-UHFFFAOYSA-N 0.000 description 1
- 229910052782 aluminium Inorganic materials 0.000 description 1
- 239000003263 anabolic agent Substances 0.000 description 1
- 230000037005 anaesthesia Effects 0.000 description 1
- 239000000729 antidote Substances 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 238000010876 biochemical test Methods 0.000 description 1
- 239000000090 biomarker Substances 0.000 description 1
- 229960002685 biotin Drugs 0.000 description 1
- 235000020958 biotin Nutrition 0.000 description 1
- 239000011616 biotin Substances 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- FAPWYRCQGJNNSJ-UBKPKTQASA-L calcium D-pantothenic acid Chemical compound [Ca+2].OCC(C)(C)[C@@H](O)C(=O)NCCC([O-])=O.OCC(C)(C)[C@@H](O)C(=O)NCCC([O-])=O FAPWYRCQGJNNSJ-UBKPKTQASA-L 0.000 description 1
- JUNWLZAGQLJVLR-UHFFFAOYSA-J calcium diphosphate Chemical compound [Ca+2].[Ca+2].[O-]P([O-])(=O)OP([O-])([O-])=O JUNWLZAGQLJVLR-UHFFFAOYSA-J 0.000 description 1
- 229960002079 calcium pantothenate Drugs 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000013339 cereals Nutrition 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000007882 cirrhosis Effects 0.000 description 1
- 208000019425 cirrhosis of liver Diseases 0.000 description 1
- AGVAZMGAQJOSFJ-WZHZPDAFSA-M cobalt(2+);[(2r,3s,4r,5s)-5-(5,6-dimethylbenzimidazol-1-yl)-4-hydroxy-2-(hydroxymethyl)oxolan-3-yl] [(2r)-1-[3-[(1r,2r,3r,4z,7s,9z,12s,13s,14z,17s,18s,19r)-2,13,18-tris(2-amino-2-oxoethyl)-7,12,17-tris(3-amino-3-oxopropyl)-3,5,8,8,13,15,18,19-octamethyl-2 Chemical compound [Co+2].N#[C-].[N-]([C@@H]1[C@H](CC(N)=O)[C@@]2(C)CCC(=O)NC[C@@H](C)OP(O)(=O)O[C@H]3[C@H]([C@H](O[C@@H]3CO)N3C4=CC(C)=C(C)C=C4N=C3)O)\C2=C(C)/C([C@H](C\2(C)C)CCC(N)=O)=N/C/2=C\C([C@H]([C@@]/2(CC(N)=O)C)CCC(N)=O)=N\C\2=C(C)/C2=N[C@]1(C)[C@@](C)(CC(N)=O)[C@@H]2CCC(N)=O AGVAZMGAQJOSFJ-WZHZPDAFSA-M 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 231100000263 cytotoxicity test Toxicity 0.000 description 1
- 238000013461 design Methods 0.000 description 1
- 235000019821 dicalcium diphosphate Nutrition 0.000 description 1
- 229910000393 dicalcium diphosphate Inorganic materials 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000013325 dietary fiber Nutrition 0.000 description 1
- 235000015872 dietary supplement Nutrition 0.000 description 1
- ZPWVASYFFYYZEW-UHFFFAOYSA-L dipotassium hydrogen phosphate Chemical compound [K+].[K+].OP([O-])([O-])=O ZPWVASYFFYYZEW-UHFFFAOYSA-L 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 239000007884 disintegrant Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 229940079593 drug Drugs 0.000 description 1
- 239000003814 drug Substances 0.000 description 1
- 235000013399 edible fruits Nutrition 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 102000015694 estrogen receptors Human genes 0.000 description 1
- 108010038795 estrogen receptors Proteins 0.000 description 1
- 238000009164 estrogen replacement therapy Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000011790 ferrous sulphate Substances 0.000 description 1
- 235000003891 ferrous sulphate Nutrition 0.000 description 1
- 238000011049 filling Methods 0.000 description 1
- 229960000304 folic acid Drugs 0.000 description 1
- 235000019152 folic acid Nutrition 0.000 description 1
- 239000011724 folic acid Substances 0.000 description 1
- 235000012631 food intake Nutrition 0.000 description 1
- 230000006870 function Effects 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- WIGCFUFOHFEKBI-UHFFFAOYSA-N gamma-tocopherol Natural products CC(C)CCCC(C)CCCC(C)CCCC1CCC2C(C)C(O)C(C)C(C)C2O1 WIGCFUFOHFEKBI-UHFFFAOYSA-N 0.000 description 1
- 239000007903 gelatin capsule Substances 0.000 description 1
- 235000006539 genistein Nutrition 0.000 description 1
- TZBJGXHYKVUXJN-UHFFFAOYSA-N genistein Natural products C1=CC(O)=CC=C1C1=COC2=CC(O)=CC(O)=C2C1=O TZBJGXHYKVUXJN-UHFFFAOYSA-N 0.000 description 1
- 229940045109 genistein Drugs 0.000 description 1
- ZCOLJUOHXJRHDI-CMWLGVBASA-N genistein 7-O-beta-D-glucoside Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC(O)=C2C(=O)C(C=3C=CC(O)=CC=3)=COC2=C1 ZCOLJUOHXJRHDI-CMWLGVBASA-N 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 230000002414 glycolytic effect Effects 0.000 description 1
- 229930182470 glycoside Natural products 0.000 description 1
- 150000002338 glycosides Chemical class 0.000 description 1
- 239000003163 gonadal steroid hormone Substances 0.000 description 1
- 208000019622 heart disease Diseases 0.000 description 1
- 230000002440 hepatic effect Effects 0.000 description 1
- 206010019692 hepatic necrosis Diseases 0.000 description 1
- 230000007866 hepatic necrosis Effects 0.000 description 1
- 230000002443 hepatoprotective effect Effects 0.000 description 1
- 230000013632 homeostatic process Effects 0.000 description 1
- PFOARMALXZGCHY-UHFFFAOYSA-N homoegonol Natural products C1=C(OC)C(OC)=CC=C1C1=CC2=CC(CCCO)=CC(OC)=C2O1 PFOARMALXZGCHY-UHFFFAOYSA-N 0.000 description 1
- 230000003054 hormonal effect Effects 0.000 description 1
- 108091008039 hormone receptors Proteins 0.000 description 1
- 230000000260 hypercholesteremic effect Effects 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 description 1
- 229910000359 iron(II) sulfate Inorganic materials 0.000 description 1
- 229960002725 isoflurane Drugs 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000003780 keratinization Effects 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 235000021374 legumes Nutrition 0.000 description 1
- 208000032839 leukemia Diseases 0.000 description 1
- 229930013686 lignan Natural products 0.000 description 1
- 235000009408 lignans Nutrition 0.000 description 1
- 150000005692 lignans Chemical class 0.000 description 1
- 208000019423 liver disease Diseases 0.000 description 1
- 231100000835 liver failure Toxicity 0.000 description 1
- 208000007903 liver failure Diseases 0.000 description 1
- 210000005228 liver tissue Anatomy 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- ZLNQQNXFFQJAID-UHFFFAOYSA-L magnesium carbonate Chemical compound [Mg+2].[O-]C([O-])=O ZLNQQNXFFQJAID-UHFFFAOYSA-L 0.000 description 1
- 239000001095 magnesium carbonate Substances 0.000 description 1
- 229910000021 magnesium carbonate Inorganic materials 0.000 description 1
- 229910001629 magnesium chloride Inorganic materials 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- DKXULEFCEORBJK-UHFFFAOYSA-N magnesium;octadecanoic acid Chemical compound [Mg].CCCCCCCCCCCCCCCCCC(O)=O DKXULEFCEORBJK-UHFFFAOYSA-N 0.000 description 1
- 230000036210 malignancy Effects 0.000 description 1
- 239000011159 matrix material Substances 0.000 description 1
- 238000000691 measurement method Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- HEBKCHPVOIAQTA-UHFFFAOYSA-N meso ribitol Natural products OCC(O)C(O)C(O)CO HEBKCHPVOIAQTA-UHFFFAOYSA-N 0.000 description 1
- 230000002503 metabolic effect Effects 0.000 description 1
- 230000007935 neutral effect Effects 0.000 description 1
- 229960003966 nicotinamide Drugs 0.000 description 1
- 235000005152 nicotinamide Nutrition 0.000 description 1
- 239000011570 nicotinamide Substances 0.000 description 1
- 231100000065 noncytotoxic Toxicity 0.000 description 1
- 230000002020 noncytotoxic effect Effects 0.000 description 1
- 231100000252 nontoxic Toxicity 0.000 description 1
- 230000003000 nontoxic effect Effects 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 150000002482 oligosaccharides Chemical class 0.000 description 1
- 210000000963 osteoblast Anatomy 0.000 description 1
- KHPXUQMNIQBQEV-UHFFFAOYSA-N oxaloacetic acid Chemical compound OC(=O)CC(=O)C(O)=O KHPXUQMNIQBQEV-UHFFFAOYSA-N 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000004792 oxidative damage Effects 0.000 description 1
- 235000020232 peanut Nutrition 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 210000002824 peroxisome Anatomy 0.000 description 1
- 230000001766 physiological effect Effects 0.000 description 1
- 230000036470 plasma concentration Effects 0.000 description 1
- 239000002574 poison Substances 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 239000004810 polytetrafluoroethylene Substances 0.000 description 1
- 229920001343 polytetrafluoroethylene Polymers 0.000 description 1
- 235000020777 polyunsaturated fatty acids Nutrition 0.000 description 1
- 239000001508 potassium citrate Substances 0.000 description 1
- 229960002635 potassium citrate Drugs 0.000 description 1
- QEEAPRPFLLJWCF-UHFFFAOYSA-K potassium citrate (anhydrous) Chemical compound [K+].[K+].[K+].[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O QEEAPRPFLLJWCF-UHFFFAOYSA-K 0.000 description 1
- 235000011082 potassium citrates Nutrition 0.000 description 1
- 239000000955 prescription drug Substances 0.000 description 1
- 238000012545 processing Methods 0.000 description 1
- 230000002250 progressing effect Effects 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 239000008213 purified water Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000001850 reproductive effect Effects 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 229960002477 riboflavin Drugs 0.000 description 1
- SEBFKMXJBCUCAI-HKTJVKLFSA-N silibinin Chemical compound C1=C(O)C(OC)=CC([C@@H]2[C@H](OC3=CC=C(C=C3O2)[C@@H]2[C@H](C(=O)C3=C(O)C=C(O)C=C3O2)O)CO)=C1 SEBFKMXJBCUCAI-HKTJVKLFSA-N 0.000 description 1
- 235000017700 silymarin Nutrition 0.000 description 1
- 229960004245 silymarin Drugs 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 238000013223 sprague-dawley female rat Methods 0.000 description 1
- 239000003381 stabilizer Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000000758 substrate Substances 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 125000003831 tetrazolyl group Chemical group 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 210000002303 tibia Anatomy 0.000 description 1
- 238000013417 toxicology model Methods 0.000 description 1
- 235000021122 unsaturated fatty acids Nutrition 0.000 description 1
- 150000004670 unsaturated fatty acids Chemical class 0.000 description 1
- 206010046766 uterine cancer Diseases 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 235000019155 vitamin A Nutrition 0.000 description 1
- 239000011719 vitamin A Substances 0.000 description 1
- 235000019163 vitamin B12 Nutrition 0.000 description 1
- 239000011715 vitamin B12 Substances 0.000 description 1
- 235000019164 vitamin B2 Nutrition 0.000 description 1
- 239000011716 vitamin B2 Substances 0.000 description 1
- 235000019154 vitamin C Nutrition 0.000 description 1
- 239000011718 vitamin C Substances 0.000 description 1
- 235000019165 vitamin E Nutrition 0.000 description 1
- 239000011709 vitamin E Substances 0.000 description 1
- 229940046009 vitamin E Drugs 0.000 description 1
- 229940045997 vitamin a Drugs 0.000 description 1
- 229940033203 vitamin b6 0.5 mg Drugs 0.000 description 1
- 238000005406 washing Methods 0.000 description 1
- 230000005186 women's health Effects 0.000 description 1
- 239000000811 xylitol Substances 0.000 description 1
- HEBKCHPVOIAQTA-SCDXWVJYSA-N xylitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)CO HEBKCHPVOIAQTA-SCDXWVJYSA-N 0.000 description 1
- 235000010447 xylitol Nutrition 0.000 description 1
- 229960002675 xylitol Drugs 0.000 description 1
- 239000011787 zinc oxide Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K36/00—Medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicines
- A61K36/18—Magnoliophyta (angiosperms)
- A61K36/185—Magnoliopsida (dicotyledons)
- A61K36/48—Fabaceae or Leguminosae (Pea or Legume family); Caesalpiniaceae; Mimosaceae; Papilionaceae
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/105—Plant extracts, their artificial duplicates or their derivatives
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/34—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having five-membered rings with one oxygen as the only ring hetero atom, e.g. isosorbide
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/33—Heterocyclic compounds
- A61K31/335—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin
- A61K31/35—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom
- A61K31/352—Heterocyclic compounds having oxygen as the only ring hetero atom, e.g. fungichromin having six-membered rings with one oxygen as the only ring hetero atom condensed with carbocyclic rings, e.g. methantheline
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/08—Solutions
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/20—Pills, tablets, discs, rods
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K9/00—Medicinal preparations characterised by special physical form
- A61K9/48—Preparations in capsules, e.g. of gelatin, of chocolate
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/16—Drugs for disorders of the alimentary tract or the digestive system for liver or gallbladder disorders, e.g. hepatoprotective agents, cholagogues, litholytics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P15/00—Drugs for genital or sexual disorders; Contraceptives
- A61P15/12—Drugs for genital or sexual disorders; Contraceptives for climacteric disorders
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/08—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease
- A61P19/10—Drugs for skeletal disorders for bone diseases, e.g. rachitism, Paget's disease for osteoporosis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2200/00—Function of food ingredients
- A23V2200/30—Foods, ingredients or supplements having a functional effect on health
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K2236/00—Isolation or extraction methods of medicinal preparations of undetermined constitution containing material from algae, lichens, fungi or plants, or derivatives thereof, e.g. traditional herbal medicine
- A61K2236/10—Preparation or pretreatment of starting material
- A61K2236/19—Preparation or pretreatment of starting material involving fermentation using yeast, bacteria or both; enzymatic treatment
Definitions
- the present invention relates to cousterol-containing germinated fermented soybean extract having a hepatocellular protective effect and an effect of increasing bone density and a composition comprising the same. More specifically, the in vitro antioxidant activity, cell viability, hematological test of animal models, MDA (malone) for the hepatotoxicity of menopausal disorders using germinated fermented soybean extract as a component that improves lipids and menopausal disorders in natural foods Dialdehyde), GST (glutathione S-transferase), GSH (reduced glutathione), SOD (Superoxide dismutase), and catalase were measured to verify hepatocellular protective effects.
- MDA malone
- the germinated fermented soybean extract as a sample in vivo
- Fermented germinated soybean extract of the present invention significantly increases blood estrogen and testosterone concentrations, thereby relieving menopausal symptoms, and increasing bone mineral protection and increasing bone mineral content in vitro and in vivo. Increase, and the thickness and number of spongy bones increase
- Didzein in legumes isoflavones contain cumesterol, which regulates female hormone balance and content to prevent breast and uterine cancer, slowing menopause and alleviating many of the symptoms associated with it.
- the aging of the population is progressing very rapidly all over the world, and especially in Korea, the progress of aging is very fast compared to other developed countries.
- women's average life expectancy is increased and the intellectual level is improved as a result of the health promotion resulting from economic growth
- interest in women's menopause and postmenopausal quality of life and health improvement is also increasing.
- the main symptoms of menopause include hot flashes, anxiety, depression and nervousness. Symptoms such as cardiovascular disease, osteoporosis and memory loss are known. The cause of these symptoms is known to be caused by interrupted production and secretion of estrogen.
- hormone replacement therapy has been proposed, but there are no results on usage, dose, and safety.
- Phytoestrogen a natural food product, is a reproductive hormone-like substance that is beginning to attract attention for the prevention of menopausal symptoms and osteoporosis, and has been studied and reported to be able to substitute estrogen in postmenopausal women.
- synthetic estrogen replacement therapy is currently used to treat menopausal symptoms, most women experience side effects such as remenstruation, breast softening and weight gain, and food intake with plant-based estrogen-like substances in natural foods. Relieving menopausal symptoms will help to alleviate the side effects.
- Phytoestrogens include isoflavones and lignans and are compounds similar to those naturally occurring in plants such as beans, fruits, vegetables, peanuts and grains. Isoflavones are attracting attention as a kind of phytoestrogens that weakly bind to estrogen receptors in the body and have various physiological effects. Isoflavones extracted from soybeans exist mainly as glycosides in plants, and are absorbed into the human body to be converted into aglycone forms of genistein and dyedzein by glycosidase and have estrogenic properties.
- composition for the prevention and treatment of menopausal symptoms comprising a soybean extract containing cumestrol as an active ingredient
- a menopause containing a soybean extract containing cumestrol as an active ingredient It is the experimental result of the cell experiment (in vitro) on the composition for the prevention of diseases, and to improve the menopausal symptoms improvement and liver cell protection effect through the animal experiment (in-vivo) to confirm the cell experiment in the present invention. And improved germinated fermented soybean extract to achieve advanced methods and results.
- Patent Document 1 Korean Patent Publication No. 10-2014-0053471
- Non-Patent Document 1 Starzec, J, J, and D.F. Berger, Effects of stress and ovariectomy on the plasma cholesterol, serum triglyceride, and aortic cholesterol levels of female rats, Physiol Behav. 1986.37 (1): p. 99-104.
- Non-Patent Document 2 Ettinger, B., Overview of estrogen replacement therapy a historical perspective. Proc Soc Exp Biol Med. 217 (1): p. 2-5.
- Non-Patent Document 3 3. Baker, M.E., K.L. Medlock, and D.M. Sheehan, Flavonoids inhibit estrogen binding to rat alpha-fetoprotein. Proc Soc Exp Biol Med. 1998.217 (3): p.317-21.
- Non-Patent Document 4 Boker, L.K., et al., Intake of Dietary Phytoestrogens by Dutch women. J Nutr. 2002. 132 (6): p. 1319-28.
- Non-Patent Document 5 Morito, K., et al., Interaction of Phytoestrogens with estrogen receptors alpha and beta, Biol Pharm Bull. 2001. 24 (4); p.6-11.
- Non-Patent Document 6 6. Albertazzi, P., et al., The effect of dietary soy supplementation on hot flushes, Obstet Gynecol. 1988. 91 (3): p. 389-94.
- Non-Patent Document 7 7. Han, K, K., et al., Benefits of soy isoflavone therapeutic regimen on menopausal symptoms, Obstet Gynecol. 2002. 99 (3): p. 389-94.
- Non-Patent Document 8 Petri Nahas, E., et al., Benefits of soy germ isoflavones in postmenopausal women with contraindication for conventional hormone replacement therapy, Maturitas. 2004. 48 (4): p.372-80.
- Non-Patent Document 9 Kaari, C., ey al., Randomized clinical trial comparing conjugated equine estrogens and isoflavones in postmenopausal women: a pilot study, Maturitus. 2006. 53 (1): p. 49-58.
- Non-Patent Document 10 Cheng, G., et al., Isoflavone treatment for acute menopausal symptoms, Menopause. 2007.14 (3Pt1): p.468-73.
- Non-Patent Document 11 DJ., Et al., Effect of soy protein foods on low-density lipoprotein oxidation and ex vivo sex hormone receptor activity--a controlled crossover trial. Metabolism. 2000.49 (4): p.537-43.
- Nonpatent Document 12 Kotsopoulos, D., et al., The effects of soy protein containing phytoesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.55 (21): p. 8589-95.
- Non-Patent Document 13 Wangen, K.E., et al., Soy Isoflavones improve plasma lipids in normocholesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.73 (2): p.225-31.
- Non-Patent Document 14 Feng, S., et al., Fungal-stressed germination of black soybeans leads to generation of oxooctadecadienoic acids in addition to glyceollins, J Agric Food Chem. 2007.55 (21): p.8589-95.
- Non-Patent Document 15 15. Rosalki, S.B. and A.Y.Foo. Two new methods for seperating and quantifying bone and liver alkaline phosphatase isoenzymes in plasma, Cli Chem. 1984. 30 (7): p.1182-6.
- the present invention uses fermented germinated soybean extract to relieve menopausal symptoms, to protect the liver cells, and to increase the mineral content of bone.
- menopause As women's average life expectancy is increased and the intellectual level is improved as a result of the health promotion resulting from economic growth, interest in women's menopause and postmenopausal quality of life and health improvement is also increasing.
- the main symptoms of menopause include hot flashes, anxiety, depression and nervousness. Symptoms such as cardiovascular disease, osteoporosis and memory loss are known. The cause of these symptoms is known to be caused by interrupted production and secretion of estrogen.
- hormone replacement therapy As a preventive measure, hormone replacement therapy has been proposed, but there are no results on usage, dose, and safety. In addition, there is a lack of appropriate drugs or health foods that have hepatoprotective effect and bone mineral density increase effect in postmenopausal women.
- the present invention relates to a cousterol-containing germinated fermented soybean extract having an effect of protecting liver cells and increasing bone density in postmenopausal women and a composition comprising the same.
- the germinated fermented soybean extract containing cumesterol to verify the effect of hepatocellular protection and bone density increase in postmenopausal women.
- the germinated fermented soybean extract as a sample in vivo
- blood calcium and phosphorus concentrations, ALP activity, bone mineral content, and bone mineral content were measured using Micro-CT.
- the present invention was found to be effective for the protection of hepatocytes by measuring in vitro antioxidant activity, cell viability and hematological tests, MDA, GST, GSH, SOD, CAT, etc. for hepatotoxicity in menopausal disorders. It was also confirmed that the administration of germinated fermented soybeans increased the mineral content of bone. Bone density was significantly increased in the rats treated with germinated fermented soybean, and the thickness and number of spongy bones were increased. Significant increases in blood estrogen levels can be expected to replace the role of estrogens in germinated fermented soybeans, thereby alleviating menopausal symptoms.
- the administration of germinated fermented soybean is expected to help improve menopausal obesity, and the high concentration of germinated fermented soybean is similar to that of estrogen. It was confirmed that there is a preventive effect of osteoporosis.
- the oxidative damage of the body to oxidative stress which is the cause of various diseases in humans, requires antioxidant enzymes such as catalase, SOD, GSH, and GST.
- antioxidant enzymes such as catalase, SOD, GSH, and GST.
- As a result of measuring liver enzyme antioxidant activity using an animal model with artificial liver damage using germinated fermented soybean it was significantly reduced and increased compared to liver damage model administered only t- BHP such as SOD, GSH, GST, and MDA. Indicated.
- the germinated fermented soybean was found to have a liver protective effect against oxidative stress.
- Figure 1 shows the weight gain of the group receiving the estrogen.
- Figure 2 shows serum lipids, cholesterol and glucose concentrations
- Figure 4 shows the bone metabolic index measurement results.
- Figure 5 shows the results of bone salt measurement.
- Figure 7 shows a 2D image taken by using the Micro-CT.
- Figure 8 shows the separation process using sep-pak of 70% ethanol germinated fermented soybeans.
- Figure 9 shows the cell viability of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.
- Figure 10 shows the growth rate of t- BHP cells of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.
- Figure 11 shows the results of ROS measurement of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.
- Figure 12 shows the results of animal model hematology test AST (GOT), ALT (GPT).
- Figure 13 shows the results of MDA measurement in Liver of the group administered t- BHP after the administration of germinated fermented soybean.
- Figure 14 shows the results of GST measurement in Liver of the group administered t- BHP after administration of germinated fermented soybeans.
- Figure 15 shows the results of GSH measurement in Liver of the group administered t- BHP after administration of germinated fermented soybean.
- Figure 16 shows the results of SOD measurement in Liver of the group administered t- BHP after the administration of germinated fermented soybean.
- Figure 17 shows the results of CAT measurements in Liver of the group administered t- BHP after the administration of germinated fermented soybeans.
- Germinated fermented soybean used in this experiment was provided by Hubio. Soybeans obtained from Jeju Agricultural Research and Extension Services are germinated and fermented by inoculation with Aspergillus oryzae according to previous studies. After the extraction process using water washing, choice and ultrasonic, it was dried and used as a test sample.
- the experimental animals were purchased from Korea Biolink (Daejeon), 8-week-old Sprague-Dawley female rats, and were bred as solid feed (Purina) in the laboratory. Therefore, the average body weight of each group was 289.72 ⁇ 0.86 g, and 7 animals of 6 groups (Negative control, Non-ovarian control group (sham), Estrogen group (OVX-estrogen), Germinated fermented soybean extract 0.5 g / kg Administered group (OVX-0.5), germinated fermented soybean extract 1.0 g / kg administered group (OVX-1.0) and germinated fermented soybean extract 2.0 g / kg administered group (OVX-2.0)].
- 6 groups Negative control, Non-ovarian control group (sham), Estrogen group (OVX-estrogen), Germinated fermented soybean extract 0.5 g / kg Administered group (OVX-0.5), germinated fermented soybean extract 1.0 g / kg administered group (
- the experimental animals were kept in a laboratory temperature of 22-24 ° C, humidity 60 ⁇ 5% using a breeding cage (20 ⁇ 26 ⁇ 13 cm), and the day and night cycle (12 hours light / 12 hours dark) was controlled by an automatic control device. It was raised in the university animal room and approved by Korea University's Animal Experiment Ethics Committee, and managed according to the 'Guide for the care and use of Laboratory Animals' (NRC). Water and diet were freely supplied, and the experimental samples were dissolved in tertiary distilled water and orally administered 3 ml daily, and the control group (Negative-control, sham) was orally administered daily with the same dose of tertiary distilled water.
- NRC Laboratory Animals'
- ovarian resection was performed by dividing the group by randomized complete block design according to body weight. Surgery was performed with isoflurane and after anesthesia, the dorsal flank was incised to remove the ovary and the incision was closed. In the case of non-ovarian resection control (Sham), fat was removed from the ovary in the same way and the incision was closed. Samples were administered orally after having a recovery period of 2 weeks after surgery.
- TG triglyceride
- TCHO total cholesterol
- HDL high density lipoprotein cholesterol
- LDL low density lipoprotein cholesterol
- GLU glucose
- Plasma levels of estradiol and testosterone were measured by the rat estradiol Elisa kit (Enzo life science, NY, USA) and rat testosterone Elisa kit (Enzo life science, NY, USA) by ELISA.
- Plasma calcium and phosphorus concentrations and alkaline alkaline phosphatase (ALP) concentrations were measured by FUJI DRI-CHEM 3500 and BONE-alkaline phosphatase (BONE-ALP) as bone markers.
- the femur was dried and crushed through the fat removal process, and then the bone salt in the femur was measured.
- a 2 mm high part of the tibia of the sacrificial rats was photographed using Micro-CT (Skyscan1072, SKYSCAN, Antwerpen, Belgium).
- the imaging tube voltage was 50 kVp
- the tube current was 500 ⁇ A
- 0.5 mm aluminum filtration
- the imaging time was 210 ms
- the pixel size was 18 ⁇ m.
- the threshold value was fixed at 58 to separate the bone column and bone marrow from the individual images, and the 3D image was reconstructed.
- Bone microstructure indicators were analyzed using Skyscan TM CT-analyzersoftware from images taken with Micro-CT.
- the indicators analyzed were Percent bone volume (BV / TV (%)), trabecular number (Tb.N ()), trabecular thickness (Tb.Th ( ⁇ m)), trabecular separation (Tb.Sp ( ⁇ m)), trabecular pattern factor (Tb.pf ()), and structure model index (SMI).
- the data obtained were processed using the statistical package for social science (version 12.0) statistical program. T-test was performed on the negative-control group to identify the significant differences among the groups. The measured values are expressed as average standard error, and p ⁇ 0.05, p ⁇ 0.01, and p ⁇ 0.001 are indicated by *, **, and *** , respectively.
- the weight gain of the estrogen-treated group was 12.1 ⁇ 8.7g, which tended to decrease relatively, but there was no significant difference in the germinated fermented soybean group. There was no significant difference in dietary intake changes.
- the ovarian resection control group, non-ovarian resection control group, and the estrogen-administered group were labeled as Neg.C, Sham, and Estradiol, and the germinated fermented soybean 0.5g / kg, 1.0g / kg, and 2.0g / kg groups were 0.5, 1.0, and 2.0, respectively. Notation.
- the figures are expressed as mean ⁇ standard error.
- estrogen secretion decreases rapidly in the ovarian resection group. At this time, the body accumulates a lot of fat cells capable of producing estrogen in order to enhance the reduced estrogen secretion, which leads to menopausal obesity. Lipid, cholesterol, and glucose concentrations in blood were identified as indicators of menopausal obesity, and the results are shown in FIG. 2.
- Neutral fat levels of the groups administered with 0.5 g / kg, 1.0 g / kg and 2.0 g / kg of germinated fermented soybean were 62.5 ⁇ 21.3 mg / dl, 59.0 ⁇ 8.5 mg / dl and 55.8 ⁇ 8.8 mg / dl, respectively.
- the decrease was similar to that of the non-ovarian ablation control 66.8 ⁇ 12.6 mg / dl.
- the blood glucose concentration was also 98.3 ⁇ 16.1 mg / dl, 123.3 ⁇ 14.4 mg / dl, 112.8 ⁇ 6.6 mg / dl in all groups administered with germinated fermented soybeans, the blood glucose concentration of 162.7 ⁇ It was confirmed that the tendency to decrease compared to 18.6 mg / dl.
- the administration of germinated fermented soybean is expected to help improve menopausal obesity.
- GOT and GPT are normal enzymes made in the liver. When the liver is damaged, these cells are broken down and the enzymes are released into the blood, increasing more than normal, so they are used as representative indicators of liver toxicity. Looking at Table 1 below, serum GOT, GPT activity analysis does not have a significant difference between groups. Therefore, it can be seen that the treatment of the sample does not show liver toxicity.
- the cause of various menopausal symptoms is a decrease in estrogen production and secretion due to aging of the ovary. Therefore, it is important to confirm the increase of blood estrogen concentration in order to alleviate the menopausal symptoms of the administration of germinated fermented soybeans.
- the estrogen concentration of the ovarian resection control group was 500.3 ⁇ 27.8 pg / ml compared to the estrogen concentration of 1020. ⁇ 63.6 pg / ml of the non-ovarian resection group, resulting in a significant (p ⁇ 0.001) decrease in estrogen concentration. You can see that. In the estrogen-administered group, it was confirmed that blood estrogen concentration was increased to 1134.1 ⁇ 205.8 pg / ml, which is similar to that of the non-ovarian ablation control group (FIG. 3).
- Testosterone is a representative anabolic hormone that is involved in muscle and fat metabolism. Lower levels of these hormones reduce muscle mass in the body, which leads to lower basal metabolism and easier obesity. It also induces skin keratinization, one of the typical symptoms of menopause.
- the concentration of testosterone in blood was decreased in all the groups that underwent ovarian ablation compared to the non-ovarian ablation control group. Among them, the testosterone concentrations of 2797.8 ⁇ 517.3 pg / ml and 1969.8 ⁇ 164.2 pg / ml were found in the estrogen-administered group and 1.0 g / kg of germinated fermented soybean, respectively. It was confirmed that the results were significantly increased (p ⁇ 0.05) compared to the control group.
- Alkaline phosphatase is a biomarker of osteoblasts involved in bone metabolism. In other diseases, such as osteoporosis, in which bone metabolism increases rapidly, the level of ALP also increases.
- the blood ALP level of the ovarian control group was 454.0 ⁇ 69.9 U / L, which was higher than that of the non-ovarian control group (340.2 ⁇ 33.31 U / L).
- the group administered 0.5 g / kg of germinated fermented soybeans 260.0 ⁇ 646.9 U / L, significantly lower ALP activity than the control group (p ⁇ 0.05).
- the Bone-ALP level was 140.8 ⁇ 58.5 U / L. In the group administered with 0.5 g / kg of germinated fermented soybean compared to the control group of ovarian resection, indicating that it was significantly lower (p ⁇ 0.05). .
- the bone inflammation was significantly lower (p ⁇ 0.05) in the ovarian resection group compared with the non-ovarian resection control group.
- the bone salts were 61.3 ⁇ 0.36% and 60.8 ⁇ 0.31%, respectively. This is a significant (p ⁇ 0.05) increase in bone salt compared to ovarian resection control. This confirmed that the germination of fermented soybeans increased the mineral content of bone.
- the ovarian control group showed a low value of 1.17 ⁇ 0.09 mm -1 , but increased to 1.69 ⁇ 0.15 mm -1 similar to that of the estrogen group in the 2.0 g / kg germinated fermented soybean group. It could be confirmed (p ⁇ 0.05).
- the ovarian control group showed a low value of 0.12 ⁇ 0.00 mm ⁇ 1 , but Tb.Th was significantly increased at all concentrations of estrogen and germinated fermented soybeans (FIG. 6). This means that the bone density of the germinated fermented soybeans increased significantly, and the thickness and number of spongy bones increased.
- the MTT assay is based on the principle that the yellow soluble tetraz olium salt is reduced to the blue insoluble formazan product by mitochondrial succina dehydrogenase in living cells.
- the absorbance of formazan is maximized at a wavelength of 540 nm.
- the absorbance measured at this wavelength reflects the concentration of living and metabolically active cells and is used as an indicator of mitochondrial activity.
- MTT is a test for measuring the mitochondrial activity to determine how toxic to a test substance by cytotoxicity test. If the test substance is toxic and the mitochondria are destroyed, the absorbance value is low.
- ROS Reactive oxygen species
- DCFH-DA Dichlorofluorescin diacetate
- ROS free radicals
- 70% EtoH germinated fermented soybean extract was incubated for 12 hours in a non-cytotoxic concentration and t- BHP 1 mM was treated for 1 hour to induce oxidative stress. . After 25 ⁇ M of DCFH-DA was added for 60 minutes, the fluorescence of DCF was measured.
- FIG. 1 compared with Normal (100 ⁇ 1.00%) fluorescence increased by (267.62 ⁇ 2.68%) by t-BHP treatment, 0.5 (195.24 ⁇ 1.95%) by 70% EtoH germinated fermented soybean extract pretreatment , 0.25mg / mL (195.24 ⁇ 1.95), 0.125mg / mL (240 ⁇ 2.41), 0.0625mg / mL (252.80 ⁇ 2.53), significantly inhibited the concentration from the highest concentration to 70% EtoH germinated fermented soybean extract It was confirmed that the active oxygen reduction effect by.
- LDH is a glycolytic enzyme and widely distributed in each tissue in the body. Especially, LDH in serum, liver, kidney, and muscle increases in heart disease, liver disease, malignancy, and leukemia, but has low specificity. In acute hepatitis, it escapes from hepatocytes like AST and ALT and increases in early acute hepatitis. In chronic hepatitis and cirrhosis, it increases to normal or mild. Significant differences were found in AST and ALT, one of the indicators of liver failure.
- ALT and AST are hepatocellular leak enzymes, which are used for hepatotoxicity studies because enzymes are released into the blood as hepatic necrosis and hepatic tissue destruction progress. 12 shows the results of measuring ALT and AST activity.
- the concentration of AST was 0.5 g / kg (83.20 ⁇ 2.86 U / L) and 1 g / kg (81.80 ⁇ 6.72 U / L), but there was no significant change in the concentration of germinated fermented soybeans. Significantly lower compared to the group (94.25 ⁇ 7.37 U / L).
- the ALT showed no significant change in the concentration of germinated fermented soybean at 0.5 g / kg (33.71 ⁇ 5.59 U / L) and 1 g / kg (38.17 ⁇ 4.49 U / L), but only the t-BHP was treated in the sample treatment group. Significantly lower compared to (48.25 ⁇ 16.19 U / L).
- Lipid peroxidation refers to a phenomenon in which polyunsaturated fatty acids in cell membranes are peroxidated by various free radicals derived from superoxide anion radicals that can occur naturally in a living body. Unsaturated fatty acids are degraded through peroxidation to produce MDA, which is an indicator of lipid peroxidation. 13, in the case of liver tissue, the group administered t- BHP after the administration of germinated fermented soybeans was significantly different between the groups at the sample concentration of 0.5 g / kg (34.25 ⁇ 5.80) and 1 g / kg (32.68 ⁇ 12.57). There was no difference, but it decreased as the concentration increased and decreased compared to the t- BHP-only group (44.66 ⁇ 1.94).
- GST is an enzyme that exists in almost every cell of the body and is involved in many metabolism as well as detoxification of the liver. In particular, GST catalyzes the reaction of binding an electron-affinity substrate to GSH.
- FIG. 14 GST measurements germination fermentation after administration of the beans treated group, t -BHP 0.5g / kg, 1g / kg, only one group administered t -BHP (25.68 ⁇ 4.28) Sample administration as compared to the concentration At 0.5 g / kg (33.65 ⁇ 12.97) and 1 g / kg (48.43 ⁇ 8.90), the sample concentration increased with increasing sample concentration, but significant difference with the t-BHP group was only observed at 1 g / kg.
- GSH is a very important antidote and powerful antioxidant in the human body that plays a key role in detoxification, immune function, and antioxidant function in the body.
- FIG. 13 GSH measurements germination fermentation after administration of the beans treated group, t -BHP 0.5g / kg, 1g / kg, t -BHP only administered group 21.07 ⁇ 8.33)
- Sample administration concentration of 0.5 as compared to the There was no significant increase as the sample concentration increased at g / kg (25.12 ⁇ 2.44) and 1 g / kg (24.40 ⁇ 2.52).
- there was no significant difference between the groups receiving the sample and the t- BHP group but there was a slight increase in the group receiving the sample.
- SOD is an enzyme that reduces superoxide anion (O 2 ) to H 2 O 2 , and is present in cytosol and mitochondria. Only Cu / Zn-SOD exists in the cytosol, but Mn-SOD exists in the matrix of mitochondria, and a small amount of Cu / Zn-SOD exists between the inner and outer layers. SOD measurement results (Fig.
- Catalase is an enzyme that protects the body from oxygen poison by reducing H 2 O 2 produced by the action of SOD or O 2 reduction to non-toxic H 2 O. Usually, it is present in peroxisome and in small amounts in mitochondria.
- CAT measurement results (Fig. 10) after germination fermented beans treated with the group treated with t -BHP 0.5g / kg, 1g / kg, t -BHP only one dose group (182.24 ⁇ 19.29) administration sample concentration of 0.5 as compared to the At g / kg (204.49 ⁇ 41.51) and 1 g / kg (293.19 ⁇ 61.02), the sample concentration increased significantly.
- the present invention may include one or more selected from the group consisting of excipients (disintegrants, binders, lubricants), stabilizers, emulsifiers, additives, etc., which are acceptable under the pharmacological, health functional food method, food hygiene method, in addition to the extract of germinated fermented soybean. Can be.
- the composition may be made and used for oral use such as warnings, granules, powders, tablets, or capsules.
- the dosage of the composition should be adjusted according to the criteria set forth in the relevant laws (use, purpose of use, patient's condition, age, sex, weight, prescription drug, disease).
- the formulation was prepared by mixing the germinated fermented soybean extract with excipients or carriers acceptable in the pharmacological method to prepare powders, tablets, capsules, pills, and granules as preparations.
- Germinated fermented soybean extract 100 mg, corn starch 100 mg, lactose 100 mg and stearic acid magnesium 2 mg were mixed and then compressed into tablets.
- Germinated fermented soybean extract 100 mg, corn starch 100 mg, lactose 100 mg, magnesium stearate 2 mg was mixed, and then prepared by filling in gelatin capsules.
- Foods were prepared by mixing the germinated fermented soybean extract with excipients or carriers allowed by the food hygiene or dietary supplement method.
- Germinated fermented soybean extract 1g Vitamin A 70 ⁇ g, Vitamin E 1.0 mg, Vitamin 0.13 mg, Vitamin B2 0.15 mg, Vitamin B6 0.5 mg, Vitamin B12 0.2 ⁇ g, Vitamin C 10 mg, Biotin 10 ⁇ g, Nicotinamide 1.7 mg, Folic acid 50 ⁇ g, calcium pantothenate 0.5 mg, mineral mixture appropriate amount, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium monophosphate 15 mg, calcium diphosphate 55 mg, potassium citrate 90 mg, carbonate Health food was prepared by mixing 100 mg of calcium and 24.8 mg of magnesium chloride.
- the proportion of the vitamin and minerals mixed with the germinated fermented soybean extract may be variously modified according to the formulation or the consumer layer.
- Germinated fermented soybean extract 1%, liquid fructose 0.5%, oligosaccharide 2%, sugar 2%, salt 0.5% and the balance of the purified water and subsidiary materials were combined and sterilized and then packaged to prepare a beverage.
- the ratio of sugars, dietary fiber, vitamins and minerals to the germinated fermented soybean extract may be variously modified depending on the dosage form or consumer layer.
- Germinated fermented soybean extract of the present invention significantly increases blood estrogen and testosterone concentrations, thereby alleviating menopausal symptoms, increasing bone mineral content, significantly increasing bone density, and the thickness and number of sponges Increases. Therefore, there is industrial applicability.
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Pharmacology & Pharmacy (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Animal Behavior & Ethology (AREA)
- Epidemiology (AREA)
- Natural Medicines & Medicinal Plants (AREA)
- Engineering & Computer Science (AREA)
- Botany (AREA)
- Mycology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Physical Education & Sports Medicine (AREA)
- Polymers & Plastics (AREA)
- Food Science & Technology (AREA)
- Nutrition Science (AREA)
- Microbiology (AREA)
- Medical Informatics (AREA)
- Alternative & Traditional Medicine (AREA)
- Biotechnology (AREA)
- Orthopedic Medicine & Surgery (AREA)
- Rheumatology (AREA)
- Obesity (AREA)
- Hematology (AREA)
- Diabetes (AREA)
- Child & Adolescent Psychology (AREA)
- Reproductive Health (AREA)
- Endocrinology (AREA)
- Gastroenterology & Hepatology (AREA)
- Medicines Containing Plant Substances (AREA)
Abstract
The present invention relates to a germinated fermented soybean extract containing coumestrol, a preparation method therefor, and a composition containing the germinated fermented soybean extract. It can be seen that the germinated fermented soybean extract of the present invention is effective in the hepatocyte protection against hepatotoxicity through in vitro antioxidative activity and cell viability, and hematological tests and MDA, GST, GSH, SOD, and CAT measurements in animal models. In addition, the germinated fermented soybean extract of the present invention increases the mineral content in bones, thereby significantly increasing the bone density, and has an effect of increasing the thickness and number of spongy bones. Furthermore, the germinated fermented soybean extract of the present invention has an effect of relieving symptoms of menopause.
Description
본 발명은 간세포 보호 효과 및 골 밀도 증가 효과를 갖는 쿠메스테롤 함유 발아 발효콩 추출물 및 이를 포함하는 조성물에 관한 것이다. 보다 상세하게는 자연식품 중에서 체내 지질 및 갱년기 장애 개선효과가 있는 성분으로 발아 발효콩 추출물을 이용하여 갱년기 장애 시 나타나는 간독성에 대하여 in vitro 항산화활성과 세포생존율 그리고 동물모델의 혈액학적 검사, MDA(malone dialdehyde), GST(glutathione S-transferase), GSH(환원형글루타치온), SOD(Superoxide dismutase), Catalase 등의 측정을 통해 간세포 보호 효과를 검증하고자 한다. 또한, 본 발명에서는 발아 발효콩 추출물을 시료로 하여 in
vivo
실험을 통해 갱년기 장애 시 나타날 수 있는 골 손실의 개선효과를 검토하기 위하여 혈중 칼슘과 인의 농도, 골 형성 지표인 ALP 활성, 골량(Bone mineral content)을 확인하였고, 해면골의 다양한 지표를 Micro-CT를 이용하여 측정하였다.The present invention relates to cousterol-containing germinated fermented soybean extract having a hepatocellular protective effect and an effect of increasing bone density and a composition comprising the same. More specifically, the in vitro antioxidant activity, cell viability, hematological test of animal models, MDA (malone) for the hepatotoxicity of menopausal disorders using germinated fermented soybean extract as a component that improves lipids and menopausal disorders in natural foods Dialdehyde), GST (glutathione S-transferase), GSH (reduced glutathione), SOD (Superoxide dismutase), and catalase were measured to verify hepatocellular protective effects. In the present invention, the germinated fermented soybean extract as a sample in vivo In order to examine the effect of improving bone loss in menopausal disorders, we examined the concentrations of calcium and phosphorus in blood, ALP activity, bone mineral content, and bone mineral content. It measured using.
본 발명의 발효발아콩 추출물은 혈중 에스트로겐과 테스토스테론 농도가 유의적으로 증가하므로 인해 갱년기 증상을 완화시키고, in vitro 와 in vivo실험에서 간세포 보호 효과와, 골의 미네랄 함량을 증가시켜 골의 밀도가 유의적으로 증가하고, 해면골의 두께와 수가 증가한다Fermented germinated soybean extract of the present invention significantly increases blood estrogen and testosterone concentrations, thereby relieving menopausal symptoms, and increasing bone mineral protection and increasing bone mineral content in vitro and in vivo. Increase, and the thickness and number of spongy bones increase
콩류의 이소플라본 중의 다이드제인(didzein)에 쿠메스테롤 성분이 들어 있으며 여성 호르몬 균형 및 함량을 조절하여 유방암과 자궁암을 예방해 주며 폐경기를 늦추거나 이와 관련된 여러 증상을 완화시킨다.Didzein in legumes isoflavones contain cumesterol, which regulates female hormone balance and content to prevent breast and uterine cancer, slowing menopause and alleviating many of the symptoms associated with it.
인구의 고령화는 전 세계적으로 매우 급속하게 진행되고 있으며, 특히, 우리나라의 경우 고령화 진전속도가 다른 선진국과 비교하여 진행속도가 매우 빨라 큰 문제가 되고 있다. 경제성장에 따른 건강 증진의 결과로 인한 우리나라 여성의 평균수명의 연장과 지적 수준의 향상 등으로 여성의 폐경기와 폐경 후의 삶의 질 향상과 건강 증진에 대한 관심 또한 점점 고조되고 있다. 폐경의 주요 증상으로는 안면홍조, 불안증, 우울, 신경과민. 심혈관질환, 골다공증 및 기억력 감퇴 등의 증상이 대표적이며 이러한 증상의 원인은 에스트로겐의 생산과 분비가 중단되어 나타난다고 알려져 있다. 이의 예방책으로 호르몬 대체요법(estrogen replacement therapy)이 제시되고 있으나 사용법, 용량, 안전성 등에 대한 결과는 없다.The aging of the population is progressing very rapidly all over the world, and especially in Korea, the progress of aging is very fast compared to other developed countries. As women's average life expectancy is increased and the intellectual level is improved as a result of the health promotion resulting from economic growth, interest in women's menopause and postmenopausal quality of life and health improvement is also increasing. The main symptoms of menopause include hot flashes, anxiety, depression and nervousness. Symptoms such as cardiovascular disease, osteoporosis and memory loss are known. The cause of these symptoms is known to be caused by interrupted production and secretion of estrogen. As a preventive measure, hormone replacement therapy has been proposed, but there are no results on usage, dose, and safety.
천연 식품 중의 식물성 에스트로겐(Phytoestrogen)은 생식 호르몬 유사물질로 폐경기 증상 및 골다공증 예방 가능성이 주목 받기 시작하면서 폐경기 이후의 여성에게 estrogen 대체작용을 할 수 있다는 것이 많이 연구, 보고되고 있다. 현재 폐경기 증상 치료에 합성 estrogen 대체 요법이 이용되면서, 대부분의 여성들은 재 월경 현상, 유방의 연화 및 체중증가 등의 부작용을 경험하고 있고, 천연 식품 속에서의 식물성 estrogen 유사물질을 가진 식품섭취를 통한 폐경기 증상 완화는 부작용 해소에 많은 도움을 주리라 생각한다.Phytoestrogen, a natural food product, is a reproductive hormone-like substance that is beginning to attract attention for the prevention of menopausal symptoms and osteoporosis, and has been studied and reported to be able to substitute estrogen in postmenopausal women. As synthetic estrogen replacement therapy is currently used to treat menopausal symptoms, most women experience side effects such as remenstruation, breast softening and weight gain, and food intake with plant-based estrogen-like substances in natural foods. Relieving menopausal symptoms will help to alleviate the side effects.
식물성 에스트로겐은 이소플라본과 리그난을 포함하며, 콩, 과일, 채소, 땅콩, 곡물 등의 식물에서 자연적으로 발생한 에스트로겐과 유사한 화합물이다. 이소플라본은 체내 에스트로겐 수용체와 약하게 결합하여 다양한 생리 효과를 가지는 식물성 에스트로겐의 일종으로 주목 받고 있다. 콩에서 추출한 이소플라본은 식물 내에서는 주로 glycoside로 존재하며, 인체 내로 흡수되면서 glycosidase에 의해 아글리콘 형태의 제니스테인과 다이드제인으로 변화되어 estrogenic property를 갖게 된다.Phytoestrogens include isoflavones and lignans and are compounds similar to those naturally occurring in plants such as beans, fruits, vegetables, peanuts and grains. Isoflavones are attracting attention as a kind of phytoestrogens that weakly bind to estrogen receptors in the body and have various physiological effects. Isoflavones extracted from soybeans exist mainly as glycosides in plants, and are absorbed into the human body to be converted into aglycone forms of genistein and dyedzein by glycosidase and have estrogenic properties.
여성의 건강과 관련하여 이소플라본의 섭취는 기존의 에스트로겐 치료제와 달리 부작용 없이 폐경기 증상을 완화시키고, 혈청 지질 농도 및 고콜레스테롤 혈증을 보이는 여성에서 심혈관계 위험도와 혈중 지질과 산화물을 감소시키는 것으로 보고된 바 있다.Concerning women's health, ingestion of isoflavones has been reported to alleviate menopausal symptoms without side effects, unlike conventional estrogen therapy, and to reduce cardiovascular risk and blood lipids and oxides in women with serum lipid levels and hypercholesterolemia. There is a bar.
대한민국 공개특허공보 제10-2014-0053471(쿠메스트롤을 포함하는 콩 추출물을 유효성분으로 포함하는 갱년기 증상의 예방 및 치료용 조성물)은 쿠메스트롤을 포함하는 콩 추출물을 유효성분으로 포함하는 갱년기 질환의 예방 등에 대한 조성물에 관한 세포실험(in vitro)의 실험 결과이며, 본원발명에서 상기의 세포실험을 확인하고자 동물실험(in-vivo)을 통해 갱년기 증상 개선 및 간세포 보호 효과 등을 확인하고자 개량 및 개선된 발아발효콩추출물로 진보된 방법과 결과를 얻고자 하는 목적입니다.Republic of Korea Patent Publication No. 10-2014-0053471 (composition for the prevention and treatment of menopausal symptoms comprising a soybean extract containing cumestrol as an active ingredient) a menopause containing a soybean extract containing cumestrol as an active ingredient It is the experimental result of the cell experiment (in vitro) on the composition for the prevention of diseases, and to improve the menopausal symptoms improvement and liver cell protection effect through the animal experiment (in-vivo) to confirm the cell experiment in the present invention. And improved germinated fermented soybean extract to achieve advanced methods and results.
{선행기술문헌}{Prior art technical literature}
(특허문헌 1) 한국특허 공개번호 10-2014-0053471(Patent Document 1) Korean Patent Publication No. 10-2014-0053471
(비특허문헌 1)1. Starzec, J,J, and D.F. Berger, Effects of stress and ovariectomy on the plasma cholesterol, serum triglyceride, and aortic cholesterol levels of female rats, Physiol Behav. 1986.37(1): p.99-104.(Non-Patent Document 1) 1. Starzec, J, J, and D.F. Berger, Effects of stress and ovariectomy on the plasma cholesterol, serum triglyceride, and aortic cholesterol levels of female rats, Physiol Behav. 1986.37 (1): p. 99-104.
(비특허문헌 2)2. Ettinger, B., Overview of estrogen replacement therapy a historical perspective. Proc Soc Exp Biol Med. 217(1): p.2-5.(Non-Patent Document 2) 2. Ettinger, B., Overview of estrogen replacement therapy a historical perspective. Proc Soc Exp Biol Med. 217 (1): p. 2-5.
(비특허문헌 3)3. Baker, M.E.,K.L. Medlock, and D.M. Sheehan, Flavonoids inhibit estrogen binding to rat alpha-fetoprotein. Proc Soc Exp Biol Med. 1998. 217(3): p.317-21.(Non-Patent Document 3) 3. Baker, M.E., K.L. Medlock, and D.M. Sheehan, Flavonoids inhibit estrogen binding to rat alpha-fetoprotein. Proc Soc Exp Biol Med. 1998.217 (3): p.317-21.
(비특허문헌 4)4. Boker, L.K., et al., Intake of Dietary Phytoestrogens by Dutch women. J Nutr. 2002. 132(6): p.1319-28.(Non-Patent Document 4) 4. Boker, L.K., et al., Intake of Dietary Phytoestrogens by Dutch women. J Nutr. 2002. 132 (6): p. 1319-28.
(비특허문헌 5)5. Morito, K., et al., Interaction of Phytoestrogens with estrogen receptors alpha and beta, Biol Pharm Bull. 2001. 24(4); p.6-11.(Non-Patent Document 5) 5. Morito, K., et al., Interaction of Phytoestrogens with estrogen receptors alpha and beta, Biol Pharm Bull. 2001. 24 (4); p.6-11.
(비특허문헌 6)6. Albertazzi, P., et al., The effect of dietary soy supplementation on hot flushes, Obstet Gynecol. 1988. 91(3): p.389-94.(Non-Patent Document 6) 6. Albertazzi, P., et al., The effect of dietary soy supplementation on hot flushes, Obstet Gynecol. 1988. 91 (3): p. 389-94.
(비특허문헌 7)7. Han, K,K., et al., Benefits of soy isoflavone therapeutic regimen on menopausal symptoms, Obstet Gynecol. 2002. 99(3): p.389-94. (Non-Patent Document 7) 7. Han, K, K., et al., Benefits of soy isoflavone therapeutic regimen on menopausal symptoms, Obstet Gynecol. 2002. 99 (3): p. 389-94.
(비특허문헌 8)8. Petri Nahas, E., et al., Benefits of soy germ isoflavones in postmenopausal women with contraindication for conventional hormone replacement therapy, Maturitas. 2004. 48(4): p.372-80.(Non-Patent Document 8) 8. Petri Nahas, E., et al., Benefits of soy germ isoflavones in postmenopausal women with contraindication for conventional hormone replacement therapy, Maturitas. 2004. 48 (4): p.372-80.
(비특허문헌 9)9. Kaari, C., ey al., Randomized clinical trial comparing conjugated equine estrogens and isoflavones in postmenopausal women: a pilot study, Maturitus. 2006. 53(1): p.49-58.(Non-Patent Document 9) 9. Kaari, C., ey al., Randomized clinical trial comparing conjugated equine estrogens and isoflavones in postmenopausal women: a pilot study, Maturitus. 2006. 53 (1): p. 49-58.
(비특허문헌 10)10. Cheng, G., et al., Isoflavone treatment for acute menopausal symptoms, Menopause. 2007.14(3Pt1):p.468-73.(Non-Patent Document 10) Cheng, G., et al., Isoflavone treatment for acute menopausal symptoms, Menopause. 2007.14 (3Pt1): p.468-73.
(비특허문헌 11) DJ., et al., Effect of soy protein foods on low-density lipoprotein oxidation and ex vivo sex hormone receptor activity - - a controlled crossover trial. Metabolism. 2000.49(4):p.537-43.(Non-Patent Document 11) DJ., Et al., Effect of soy protein foods on low-density lipoprotein oxidation and ex vivo sex hormone receptor activity--a controlled crossover trial. Metabolism. 2000.49 (4): p.537-43.
(비특허문헌 12)12. Kotsopoulos, D., et al., The effects of soy protein containing phytoesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.55(21):p.8589-95.(Nonpatent Document 12) Kotsopoulos, D., et al., The effects of soy protein containing phytoesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.55 (21): p. 8589-95.
(비특허문헌 13)13. Wangen, K.E., et al., Soy Isoflavones improve plasma lipids in normocholesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.73(2):p.225-31.(Non-Patent Document 13) 13. Wangen, K.E., et al., Soy Isoflavones improve plasma lipids in normocholesterolemic and mildly hypercholesterolemic postmenopausal women. Am J Clin Nutr. 2001.73 (2): p.225-31.
(비특허문헌 14)14. Feng, S., et al., Fungal-stressed germination of black soybeans leads to generation of oxooctadecadienoic acids in addition to glyceollins, J Agric Food Chem. 2007.55(21):p.8589-95.(Non-Patent Document 14) 14. Feng, S., et al., Fungal-stressed germination of black soybeans leads to generation of oxooctadecadienoic acids in addition to glyceollins, J Agric Food Chem. 2007.55 (21): p.8589-95.
(비특허문헌 15)15. Rosalki, S.B. and A.Y.Foo. Two new methods for seperating and quantifying bone and liver alkaline phosphatase isoenzymes in plasma, Cli Chem. 1984. 30(7): p.1182-6.(Non-Patent Document 15) 15. Rosalki, S.B. and A.Y.Foo. Two new methods for seperating and quantifying bone and liver alkaline phosphatase isoenzymes in plasma, Cli Chem. 1984. 30 (7): p.1182-6.
본 발명은 발효발아콩 추출물을 이용하여 갱년기 증상을 완화시키고, 간세포 보호 효과와, 골의 미네랄 함량을 증가시키는데 있다.The present invention uses fermented germinated soybean extract to relieve menopausal symptoms, to protect the liver cells, and to increase the mineral content of bone.
경제성장에 따른 건강 증진의 결과로 인한 우리나라 여성의 평균수명의 연장과 지적 수준의 향상 등으로 여성의 폐경기와 폐경 후의 삶의 질 향상과 건강 증진에 대한 관심 또한 점점 고조되고 있다. 폐경의 주요 증상으로는 안면홍조, 불안증, 우울, 신경과민. 심혈관질환, 골다공증 및 기억력 감퇴 등의 증상이 대표적이며 이러한 증상의 원인은 에스트로겐의 생산과 분비가 중단되어 나타난다고 알려져 있다. 이의 예방책으로 호르몬 대체요법(estrogen replacement therapy)이 제시되고 있으나 사용법, 용량, 안전성 등에 대한 결과는 없다. 더욱이 폐경기의 여성에 간세포 보호 효과 및 골 밀도 증가 효과를 갖는 마땅한 약품이나 건강식품이 부족한 실정이다.As women's average life expectancy is increased and the intellectual level is improved as a result of the health promotion resulting from economic growth, interest in women's menopause and postmenopausal quality of life and health improvement is also increasing. The main symptoms of menopause include hot flashes, anxiety, depression and nervousness. Symptoms such as cardiovascular disease, osteoporosis and memory loss are known. The cause of these symptoms is known to be caused by interrupted production and secretion of estrogen. As a preventive measure, hormone replacement therapy has been proposed, but there are no results on usage, dose, and safety. In addition, there is a lack of appropriate drugs or health foods that have hepatoprotective effect and bone mineral density increase effect in postmenopausal women.
본 발명은 폐경기의 여성에 간세포 보호 및 골 밀도 증가 효과를 갖는 쿠메스테롤 함유 발아 발효콩 추출물 및 이를 포함하는 조성물에 관한 것이다. 상기의 쿠메스테롤을 함유한 발아 발효콩 추출물을 이용하여 폐경기의 여성에 간세포 보호 및 골 밀도 증가 효과를 검증하고자 한다. 또한, 본 발명에서는 발아 발효콩 추출물을 시료로 하여 in
vivo
실험을 통해 갱년기 장애 시 나타날 수 있는 폐경기의 여성에 간세포 보호 및 골 손실의 개선효과를 검토하기 위하여 혈중 칼슘과 인의 농도, 골 형성 지표인 ALP 활성, 골량(Bone mineral content)을 확인하였고, 해면골의 다양한 지표를 Micro-CT를 이용하여 측정하였다.The present invention relates to a cousterol-containing germinated fermented soybean extract having an effect of protecting liver cells and increasing bone density in postmenopausal women and a composition comprising the same. The germinated fermented soybean extract containing cumesterol to verify the effect of hepatocellular protection and bone density increase in postmenopausal women. In the present invention, the germinated fermented soybean extract as a sample in vivo In order to examine the effect of improving hepatocellular protection and bone loss in postmenopausal women during menopausal disorders, we examined blood calcium and phosphorus concentrations, ALP activity, bone mineral content, and bone mineral content. Various indicators were measured using Micro-CT.
혈중 에스트로겐 농도가 유의적으로 증가하는 것을 통해 발아 발효콩의 isoflavone이 에스트로겐의 역할을 대체할 수 있고 이로 인해 갱년기 증상이 완화될 것을 기대할 수 있다. 테스토스테론 농도는 난소절제 대조군에 비해 유의적으로 (p < 0.05) 증가한 결과임을 확인하였다.Significant increases in blood estrogen levels can be expected to replace the role of estrogens in germinated fermented soybeans, thereby alleviating menopausal symptoms. Testosterone concentration was found to be significantly increased (p <0.05) compared to the ovarian control group.
본 발명은 갱년기 장애 시 나타나는 간독성에 대하여 in vitro 항산화활성과 세포생존율 그리고 동물모델의 혈액학적 검사, MDA, GST, GSH, SOD, CAT등의 측정을 통해 간세포 보호에 효과적임을 알 수 있었다. 또한 발아 발효콩 투여가 골의 미네랄 함량을 증가시키는 것을 확인할 수 있었다. 발아 발효콩을 투여한 쥐에서 골의 밀도가 유의적으로 증가했고, 해면골의 두께와 수가 증가하였다는 것을 의미한다. 혈중 에스트로겐 농도가 유의적으로 증가하는 것을 통해 발아 발효콩의 isoflavone이 에스트로겐의 역할을 대체할 수 있고 이로 인해 갱년기 증상이 완화될 것을 기대할 수 있다. 특히, 혈액 내 중성지방의 농도와 포도당의 농도가 감소하는 경향을 갖는 것으로 보아, 발아 발효콩의 투여가 갱년기 비만 개선에 도움을 줄 것으로 예상되고, 고농도의 발아 발효콩 투여가 에스트로겐 투여와 비슷한 수준으로 골다공증의 예방효과가 있다는 것을 확인했다. 인간의 다양한 질병의 원인으로 지목되는 산화적 스트레스에 대한 생체의 산화적 손상은 catalase, SOD, GSH, GST와 같은 항산화 효소들이 필요하다. 이에 발아 발효콩을 이용하여 인위적 간 손상을 가진 동물모델을 가지고 간효소 항산화 활성을 측정한 결과 SOD, GSH, GST, MDA등 t-BHP만 투여한 간손상 모델에 대비하여 유의적으로 감소 및 증가를 나타냈다. 이에 발아 발효콩은 산화적 스트레스에 대한 간 보호 효과가 있음을 알 수 있었다.The present invention was found to be effective for the protection of hepatocytes by measuring in vitro antioxidant activity, cell viability and hematological tests, MDA, GST, GSH, SOD, CAT, etc. for hepatotoxicity in menopausal disorders. It was also confirmed that the administration of germinated fermented soybeans increased the mineral content of bone. Bone density was significantly increased in the rats treated with germinated fermented soybean, and the thickness and number of spongy bones were increased. Significant increases in blood estrogen levels can be expected to replace the role of estrogens in germinated fermented soybeans, thereby alleviating menopausal symptoms. In particular, since the concentration of triglycerides and glucose in blood tend to decrease, the administration of germinated fermented soybean is expected to help improve menopausal obesity, and the high concentration of germinated fermented soybean is similar to that of estrogen. It was confirmed that there is a preventive effect of osteoporosis. The oxidative damage of the body to oxidative stress, which is the cause of various diseases in humans, requires antioxidant enzymes such as catalase, SOD, GSH, and GST. As a result of measuring liver enzyme antioxidant activity using an animal model with artificial liver damage using germinated fermented soybean, it was significantly reduced and increased compared to liver damage model administered only t- BHP such as SOD, GSH, GST, and MDA. Indicated. The germinated fermented soybean was found to have a liver protective effect against oxidative stress.
도 1은 에스트로겐을 투여한 군의 체중 증가량을 나타낸 것이다.Figure 1 shows the weight gain of the group receiving the estrogen.
도 2는 혈청 지질, 콜레스테롤 및 포도당 농도를 나타낸 것이다Figure 2 shows serum lipids, cholesterol and glucose concentrations
도 3은 호르몬의 변화를 나타낸 것이다.3 shows the change in hormone.
도 4는 골대사 지표 측정결과를 나타낸 것이다.Figure 4 shows the bone metabolic index measurement results.
도 5는 골염량 측정결과를 나타낸 것이다.Figure 5 shows the results of bone salt measurement.
도 6은 골 미세 구조 지표를 나타낸 것이다.6 shows bone microstructural indicators.
도 7은 Micro-CT를 이용하여 촬영한 2D이미지를 나타낸 것이다.Figure 7 shows a 2D image taken by using the Micro-CT.
도 8은 70% 에탄올 발아 발효콩의 sep-pak을 이용한 분리과정을 나타낸 것이다.Figure 8 shows the separation process using sep-pak of 70% ethanol germinated fermented soybeans.
도 9는 발아 발효콩 및 70% 에탄올 추출물 발아 발효콩의 세포생존율을 나타낸 것이다.Figure 9 shows the cell viability of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.
도 10은 발아 발효콩 및 70% 에탄올 추출물 발아 발효콩의 t-BHP세포생존율을 나타낸 것이다.Figure 10 shows the growth rate of t- BHP cells of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.
도 11은 발아 발효콩 및 70% 에탄올 추출물 발아 발효콩의 ROS 측정 결과를 나타낸 것이다.Figure 11 shows the results of ROS measurement of germinated fermented soybean and 70% ethanol extract germinated fermented soybean.
도 12는 동물모델 혈액학적검사 AST(GOT), ALT(GPT) 측정결과를 나타낸 것이다.Figure 12 shows the results of animal model hematology test AST (GOT), ALT (GPT).
도 13은 발아 발효콩을 투여한 후 t-BHP를 투여한 군의 Liver에서의 MDA 측정결과를 나타낸 것이다.Figure 13 shows the results of MDA measurement in Liver of the group administered t- BHP after the administration of germinated fermented soybean.
도 14는 발아 발효콩을 투여한 후 t-BHP를 투여한 군의 Liver에서의 GST 측정결과를 나타낸 것이다.Figure 14 shows the results of GST measurement in Liver of the group administered t- BHP after administration of germinated fermented soybeans.
도 15는 발아 발효콩을 투여한 후 t-BHP를 투여한 군의 Liver에서의 GSH 측정결과를 나타낸 것이다.Figure 15 shows the results of GSH measurement in Liver of the group administered t- BHP after administration of germinated fermented soybean.
도 16은 발아 발효콩을 투여한 후 t-BHP를 투여한 군의 Liver에서의 SOD 측정결과를 나타낸 것이다.Figure 16 shows the results of SOD measurement in Liver of the group administered t- BHP after the administration of germinated fermented soybean.
도 17은 발아 발효콩을 투여한 후 t-BHP를 투여한 군의 Liver에서의 CAT 측정결과를 나타낸 것이다.Figure 17 shows the results of CAT measurements in Liver of the group administered t- BHP after the administration of germinated fermented soybeans.
<실시예 1>; 재료 및 방법<Example 1>; Materials and methods
1.1. 실험재료1.1. Experimental material
본 실험에 사용한 발아 발효콩은 휴바이오로부터 제공받았다. 제주 농업기술원(Jeju Agricultural Research and Extension Services)으로부터 얻은 대두를 선행연구에 따라 발아시키고 Aspergillus oryzae를 접종하여 발효시켜 얻는다. 수세, 정선 및 초음파를 이용한 추출 과정을 거친 뒤, 건조 하여 실험 시료로 사용하였다.Germinated fermented soybean used in this experiment was provided by Hubio. Soybeans obtained from Jeju Agricultural Research and Extension Services are germinated and fermented by inoculation with Aspergillus oryzae according to previous studies. After the extraction process using water washing, choice and ultrasonic, it was dried and used as a test sample.
1.2. 시료의 제조1.2. Preparation of Sample
대두는 수세, 정선, 발아 및 Aspergillus
oryzae를 이용한 발효과정을 거쳐 여과한 뒤 60˚C에서 건조하였다. 건조시료 10 g에 80% 에탄올을 7 mL/g으로 가하여 분해한 뒤, 초음파를 이용한 열수추출을 하였다. 그 후, 원심분리을 이용하여 발아 발효콩 추출물을 얻었고, 이를 PTFE membrane으로 여과한 뒤, 건조하여 시료를 얻었다. 위 시료를 3차 증류수에 녹여 동물실험에 사용하였다Soybeans wash, select, germinate and Aspergillus After the fermentation process using oryzae filtered and dried at 60˚C. 80% ethanol was added to 7 mL / g of 10 g of the dry sample, and the hydrothermal extract was extracted using ultrasonic waves. Thereafter, germinated fermented soybean extract was obtained by centrifugation, which was filtered through a PTFE membrane and dried to obtain a sample. The sample was dissolved in tertiary distilled water and used for animal experiments.
1.3. 실험동물1.3. Laboratory animals
실험동물은 8주령의 Sprague-Dawley계 암컷 흰쥐를 대한바이오링크(대전)로부터 구입하여 본 실험실에서 고형사료(Purina)로 사육하였고, 실험 시작 전 2주일 동안 대조군 식이로 적응시킨 후 동물의 체중에 따라 각 군의 평균 체중을 289.72±0.86 g이 되도록 7마리씩 6군[난소절제 대조군(Negative control), 비 난소절제 대조군(sham), 에스트로겐 투여군(OVX-estrogen), 발아 발효콩 추출물 0.5 g/kg 투여군(OVX-0.5), 발아 발효콩 추출물 1.0 g/kg 투여군(OVX-1.0) 및 발아 발효콩 추출물 2.0 g/kg 투여군(OVX-2.0)]으로 나누어 6주간 실험하였다. 실험동물은 사육케이지 (20×26×13 cm)를 이용해 실험실 온도 22-24℃, 습도 60±5%가 유지되며 밤낮 주기(12 시간 light/ 12 시간 dark)가 자동 조절 장치에 의해 조절되는 고려대학교 동물실에서 사육하며 고려대학교 동물실험 윤리위원회 승인을 받은 후 '실험동물 관리 및 이용에 관한 지침 (Guide for the care and use of Laboratory Animals, NRC)'에 맞추어 관리하면서 실시하였다. 물과 식이는 자유 공급하였고, 실험 시료는 3차 증류수에 용해하여 매일 3 ml씩 경구 투여 하였고, 대조군(Negative-control, sham)은 동일용량의 3차 증류수를 매일 경구 투여하였다.The experimental animals were purchased from Korea Biolink (Daejeon), 8-week-old Sprague-Dawley female rats, and were bred as solid feed (Purina) in the laboratory. Therefore, the average body weight of each group was 289.72 ± 0.86 g, and 7 animals of 6 groups (Negative control, Non-ovarian control group (sham), Estrogen group (OVX-estrogen), Germinated fermented soybean extract 0.5 g / kg Administered group (OVX-0.5), germinated fermented soybean extract 1.0 g / kg administered group (OVX-1.0) and germinated fermented soybean extract 2.0 g / kg administered group (OVX-2.0)]. The experimental animals were kept in a laboratory temperature of 22-24 ° C, humidity 60 ± 5% using a breeding cage (20 × 26 × 13 cm), and the day and night cycle (12 hours light / 12 hours dark) was controlled by an automatic control device. It was raised in the university animal room and approved by Korea University's Animal Experiment Ethics Committee, and managed according to the 'Guide for the care and use of Laboratory Animals' (NRC). Water and diet were freely supplied, and the experimental samples were dissolved in tertiary distilled water and orally administered 3 ml daily, and the control group (Negative-control, sham) was orally administered daily with the same dose of tertiary distilled water.
1.4. 난소 절제 시술1.4. Ovarian excision procedure
2주일 동안 주위환경에 적응시켜 체중에 따라 난괴법(Randomized Complete Block Design)에 의해 군을 나누어 난소절제 시술을 하였다. 수술은 isoflurane으로 마취 후 심마취기에 이르면 등쪽 옆구리를 절개하여 난소를 제거하고 절개부위를 봉합하였다. 비 난소절제 대조군(Sham)의 경우, 같은 방법으로 난소 옆의 지방을 제거하고 절개부위를 봉합하였다. 시료는 수술 후 2주간의 회복기를 가진 후에 경구 투여하였다.For 2 weeks, ovarian resection was performed by dividing the group by randomized complete block design according to body weight. Surgery was performed with isoflurane and after anesthesia, the dorsal flank was incised to remove the ovary and the incision was closed. In the case of non-ovarian resection control (Sham), fat was removed from the ovary in the same way and the incision was closed. Samples were administered orally after having a recovery period of 2 weeks after surgery.
1.5. 혈청 지질 농도 및 효소 활성 분석1.5. Serum Lipid Concentration and Enzyme Activity Analysis
분리한 혈청의 중성지방(TG), 총 콜레스테롤(TCHO), 고밀도지단백 콜레스테롤(HDL), 저밀도지단백 콜레스테롤(LDL), 포도당(GLU)은 FUJI DRI-CHEM 3500를 이용하여 측정하였다. Glutamate oxaloacetate transaminase(GOT), Glutamate pyruvate transmaminase (GPT) 활성 또한 같은 방법으로 측정하였다.The triglyceride (TG), total cholesterol (TCHO), high density lipoprotein cholesterol (HDL), low density lipoprotein cholesterol (LDL) and glucose (GLU) of the isolated serum were measured using FUJI DRI-CHEM 3500. Glutamate oxaloacetate transaminase (GOT) and Glutamate pyruvate transmaminase (GPT) activities were also measured in the same way.
1.6. 호르몬 측정1.6. Hormone measurement
혈장 내 estradiol, testosterone의 양은 ELISA법을 이용하여 각각 rat estradiol Elisa kit(Enzo life science, NY, USA)과 rat testosterone Elisa kit(Enzo life science, NY, USA) 로 측정하였다.Plasma levels of estradiol and testosterone were measured by the rat estradiol Elisa kit (Enzo life science, NY, USA) and rat testosterone Elisa kit (Enzo life science, NY, USA) by ELISA.
1.7. 골대사 지표 측정 1.7. Bone metabolism indicator measurement
골형성 지표로 혈장 칼슘과 인의 농도, 혈장 alkaline phosphatase(ALP)의 농도는 FUJI DRI-CHEM 3500로 측정하였으며 BONE-alkaline phosphatase(BONE-ALP)은 선행 연구에 따라 측정하였다. 대퇴골을 건조, 제지방 과정을 거쳐 회화하였고, 그 후 대퇴골의 골염량을 측정하였다.Plasma calcium and phosphorus concentrations and alkaline alkaline phosphatase (ALP) concentrations were measured by FUJI DRI-CHEM 3500 and BONE-alkaline phosphatase (BONE-ALP) as bone markers. The femur was dried and crushed through the fat removal process, and then the bone salt in the femur was measured.
1.8. 골조직 형태 측정: Micro-CT 촬영1.8. Bone Morphology Measurement: Micro-CT
희생 쥐의 경골 중 성장판으로부터 1mm 떨어진 부위를 2mm 높이 부분을 Micro-CT(Skyscan1072, SKYSCAN, Antwerpen, Belgium)를 이용하여 촬영하였다. 촬영 관전압은 50 kVp, 관전류는 500 μA, 0.5 mm 알루미늄 여과를 이용하였고, 촬영시간은 210 ms이었으며, 화소 크기(pixel size)는 18 μm였다. 개개의 영상에서 골소주와 골수강을 분리하기 위해 역치값(threshold value)을 58로 고정하였으며 3차원 영상을 재구성하였다. Micro-CT로 촬영된 영상들로부터 SkyscanTMCT-analyzersoftware가 사용되어 골 미세 구조 지표들이 분석되었다. 이용 분석된 지표들은 Percent bone volume (BV/TV (%)), trabecular number (Tb.N ()), trabecular thickness (Tb.Th (μm)), trabecular separation (Tb.Sp (μm)), trabecular pattern factor (Tb.pf ()), and structure model index (SMI) 이었다.A 2 mm high part of the tibia of the sacrificial rats was photographed using Micro-CT (Skyscan1072, SKYSCAN, Antwerpen, Belgium). The imaging tube voltage was 50 kVp, the tube current was 500 μA, 0.5 mm aluminum filtration, the imaging time was 210 ms, and the pixel size was 18 μm. The threshold value was fixed at 58 to separate the bone column and bone marrow from the individual images, and the 3D image was reconstructed. Bone microstructure indicators were analyzed using Skyscan ™ CT-analyzersoftware from images taken with Micro-CT. The indicators analyzed were Percent bone volume (BV / TV (%)), trabecular number (Tb.N ()), trabecular thickness (Tb.Th (μm)), trabecular separation (Tb.Sp (μm)), trabecular pattern factor (Tb.pf ()), and structure model index (SMI).
1.9. 통계처리1.9. Statistical processing
연구결과 얻어진 자료를 SPSS (statistical package for social science, version 12.0) 통계 프로그램을 사용하여 처리하였다. 집단간의 유의적인 차이를 알아보기 위해 Negative-control 군에 대한 T-test를 실시하였다. 측정값은 평균 표준오차로 표기하였으며, p<0.05, p<0.01, p<0.001 은 각각 *, **, *** 로 표기되었다.The data obtained were processed using the statistical package for social science (version 12.0) statistical program. T-test was performed on the negative-control group to identify the significant differences among the groups. The measured values are expressed as average standard error, and p <0.05, p <0.01, and p <0.001 are indicated by *, **, and *** , respectively.
<실시예 1> 실험결과Example 1 Experimental Results
2.1. 체중 변화 및 식이섭취량2.1. Weight change and dietary intake
6 주간 실험을 진행하는 동안, 비 난소절제 대조군에 비해 난소절제술로 갱년기를 유도한 모든 군에서 체중이 증가하는 경향을 확인했다(도 1). 비 난소절제 대조군의 체중 증가량은 20.4 ± 3.3 g 이고, 이에 비해 난소절제 대조군, 발아 발효콩을 0.5 g/kg, 1.0 g/kg, 2.0 g/kg로 투여한 군의 체중 증가량은 각각 39.8 ± 11.1 g, 42.2 ± 4.5 g, 34.0 ± 5.6 g, 36.8 ± 4.3 g 임을 확인할 수 있었다. 에스트로겐을 투여한 군의 체중 증가량은 12.1 ± 8.7g 으로, 비교적 감소하는 경향을 보였으나, 발아 발효콩 투여 군에서는 유의적인 차이가 보이지 않았다. 식이섭취량 변화에서도 유의적인 차이를 보이지 않았다. 난소절제 대조군, 비 난소절제 대조군, 에스토로겐 투여군은 각 Neg.C, Sham, Estradiol로 표기하였고, 발아 발효콩 0.5g/kg, 1.0g/kg, 2.0g/kg 투여군은 각각 0.5, 1.0, 2.0으로 표기하였다. 수치는 평균 ± 표준오차로 표기하였다.During the six-week experiment, body weight gain was observed in all groups induced by menopause by ovarian resection compared to the non-ovarian resection control (FIG. 1). The weight gain of the non-ovarian control group was 20.4 ± 3.3 g, whereas the weight gain of the ovarian control group and the germinated fermented soybean at 0.5 g / kg, 1.0 g / kg and 2.0 g / kg were 39.8 ± 11.1, respectively. g, 42.2 ± 4.5 g, 34.0 ± 5.6 g, 36.8 ± 4.3 g was confirmed. The weight gain of the estrogen-treated group was 12.1 ± 8.7g, which tended to decrease relatively, but there was no significant difference in the germinated fermented soybean group. There was no significant difference in dietary intake changes. The ovarian resection control group, non-ovarian resection control group, and the estrogen-administered group were labeled as Neg.C, Sham, and Estradiol, and the germinated fermented soybean 0.5g / kg, 1.0g / kg, and 2.0g / kg groups were 0.5, 1.0, and 2.0, respectively. Notation. The figures are expressed as mean ± standard error.
2.2. 혈청 지질 농도 및 효소 활성 분석2.2. Serum Lipid Concentration and Enzyme Activity Analysis
여성 갱년기와 마찬가지로, 난소절제를 실시한 그룹에서는 에스트로겐 분비가 급격하게 감소한다. 이때 신체는 감소한 에스트로겐 분비를 보강하기 위하여 에스트로겐을 생산할 수 있는 지방 세포를 많이 축척하게 되고 이로 인해 갱년기 비만이 유도된다. 갱년기 비만의 지표로 혈액 내의 지질, 콜레스테롤, 포도당 농도를 확인하였고 그 결과는 도 2와 같다. As in female menopause, estrogen secretion decreases rapidly in the ovarian resection group. At this time, the body accumulates a lot of fat cells capable of producing estrogen in order to enhance the reduced estrogen secretion, which leads to menopausal obesity. Lipid, cholesterol, and glucose concentrations in blood were identified as indicators of menopausal obesity, and the results are shown in FIG. 2.
발아 발효콩을 0.5 g/kg, 1.0 g/kg, 2.0 g/kg로 투여한 군의 중성 지방 수치는 각각 62.5 ± 21.3 mg/dl, 59.0 ± 8.5 mg/dl, 55.8 ± 8.8 mg/dl으로, 비 난소절제 대조군 66.8 ± 12.6 mg/dl 과 유사한 수치로 감소하는 것을 확인할 수 있었다. Neutral fat levels of the groups administered with 0.5 g / kg, 1.0 g / kg and 2.0 g / kg of germinated fermented soybean were 62.5 ± 21.3 mg / dl, 59.0 ± 8.5 mg / dl and 55.8 ± 8.8 mg / dl, respectively. The decrease was similar to that of the non-ovarian ablation control 66.8 ± 12.6 mg / dl.
총 콜레스테롤의 경우, 발아 발효콩 1.0 g/kg, 2.0 g/kg로 투여한 경우 각각 139.5 ± 9.6 mg/dl, 136.7 ± 6.5 mg/dl으로 수치가 증가한 것이 확인되었다. 결과적으로 cholesterol pool이 커져 고밀도지단백 콜레스테롤과 저밀도지단백 콜레스테롤에서도 유사한 효과가 나타난 것으로 보인다. In the case of total cholesterol, the values increased to 139.5 ± 9.6 mg / dl and 136.7 ± 6.5 mg / dl, respectively, when administered at 1.0 g / kg and 2.0 g / kg of germinated fermented soybean. As a result, the cholesterol pool was enlarged, and similar effects were observed in high density lipoprotein cholesterol and low density lipoprotein cholesterol.
도 2를 살펴보면, 혈중 포도당의 농도 또한 발아 발효콩을 투여한 모든 군에서 98.3 ± 16.1 mg/dl, 123.3 ± 14.4 mg/dl, 112.8 ± 6.6 mg/dl으로 난소절제 대조군의 혈중 포도당 농도인 162.7 ±18.6 mg/dl 에 비해 감소하는 경향이 나타나는 것을 확인할 수 있었다.Referring to Figure 2, the blood glucose concentration was also 98.3 ± 16.1 mg / dl, 123.3 ± 14.4 mg / dl, 112.8 ± 6.6 mg / dl in all groups administered with germinated fermented soybeans, the blood glucose concentration of 162.7 ± It was confirmed that the tendency to decrease compared to 18.6 mg / dl.
혈액 내 중성지방의 농도와 포도당의 농도가 감소하는 경향을 갖는 것으로 보아, 발아 발효콩의 투여가 갱년기 비만 개선에 도움을 줄 것으로 예상된다.Given that the concentration of triglycerides and glucose in the blood tend to decrease, the administration of germinated fermented soybean is expected to help improve menopausal obesity.
GOT, GPT는 간에서 만들어지는 정상 효소이다. 간이 손상 받을 경우 간세포가 깨지면서 이 효소들이 혈액으로 유리되어 정상보다 많이 증가하기 때문에 이들은 대표적인 간 독성의 지표로 사용된다. 하기 표 1을 살펴보면 혈청 GOT, GPT 활성 분석은 그룹 별로 유의적인 차이를 갖지 않는다. 따라서 시료의 처리가 간 독성을 나타내지 않는 것을 알 수 있다. GOT and GPT are normal enzymes made in the liver. When the liver is damaged, these cells are broken down and the enzymes are released into the blood, increasing more than normal, so they are used as representative indicators of liver toxicity. Looking at Table 1 below, serum GOT, GPT activity analysis does not have a significant difference between groups. Therefore, it can be seen that the treatment of the sample does not show liver toxicity.
ToxicityToxicity | ShamSham | Neg.CNeg.C | EstradiolEstradiol | 0.50.5 | 1.01.0 | 2.02.0 |
GOT(U/L)GOT (U / L) | 92.00±10.4292.00 ± 10.42 | 76.57±7.2876.57 ± 7.28 | 63.71±5.8963.71 ± 5.89 | 62.71±6.3562.71 ± 6.35 | 74.00±3.9174.00 ± 3.91 | 69.14±5.1169.14 ± 5.11 |
GPT(U/L)GPT (U / L) | 28.43±3.9028.43 ± 3.90 | 23.86±1.5823.86 ± 1.58 | 31.00±3.7231.00 ± 3.72 | 22.00±6.2522.00 ± 6.25 | 23.86±0.5923.86 ± 0.59 | 26.43±5.4226.43 ± 5.42 |
2.3. 호르몬 측정2.3. Hormone measurement
다양한 갱년기 증상의 원인은 난소의 노화로 인한 에스트로겐 생산과 분비의 감소이다. 따라서 발아 발효콩의 투여의 갱년기 증상을 완화 효과를 보기 위해서는 혈중 에스트로겐 농도 증가를 확인하는 것이 중요하다. The cause of various menopausal symptoms is a decrease in estrogen production and secretion due to aging of the ovary. Therefore, it is important to confirm the increase of blood estrogen concentration in order to alleviate the menopausal symptoms of the administration of germinated fermented soybeans.
2.3.1. 에스트로겐2.3.1. Estrogen
비 난소절제 대조군의 에스트로겐 농도 1020. ± 63.6 pg/ml와 비교하여 난소절제 대조군의 에스트로겐 농도가 500.3 ± 27.8 pg/ml인 것으로 보아, 난소절제를 통해 에스트로겐 농도가 유의적으로 (p < 0.001) 감소한 것을 확인할 수 있다. 에스트로겐 투여군의 경우, 비 난소절제 대조군과 유사한 수치인 1134.1 ± 205.8 pg/ml로 혈중 에스트로겐 농도가 증가한 것을 확인하였다(도 3). 발아 발효콩을 투여한 세 군 모두 난소절제 대조군과 비교하여 각각 1577.1 ± 341.6 pg/ml, 2358.6 ± 191.8 pg/ml, 938.5 ± 51.2 pg/ml 유의적으로 혈중 에스트로겐 농도가 증가하였다(도 3). 혈중 에스트로겐 농도가 유의적으로 증가하는 것을 통해 발아 발효콩의 쿠메스테롤을 함유한 추출물이 에스트로겐의 역할을 대체할 수 있고 이로 인해 갱년기 증상이 완화될 것을 기대할 수 있다.The estrogen concentration of the ovarian resection control group was 500.3 ± 27.8 pg / ml compared to the estrogen concentration of 1020. ± 63.6 pg / ml of the non-ovarian resection group, resulting in a significant (p <0.001) decrease in estrogen concentration. You can see that. In the estrogen-administered group, it was confirmed that blood estrogen concentration was increased to 1134.1 ± 205.8 pg / ml, which is similar to that of the non-ovarian ablation control group (FIG. 3). All three groups that received germinated fermented soybeans significantly increased blood estrogen levels in 1577.1 ± 341.6 pg / ml, 2358.6 ± 191.8 pg / ml, and 938.5 ± 51.2 pg / ml, respectively, as compared to the ovarian ablation control (FIG. 3). Significantly increased blood estrogen levels can be expected to reduce the role of estrogens by extracts containing cousterol from germinated fermented soybeans, thereby alleviating menopausal symptoms.
2.3.2. 테스토스테론2.3.2. Testosterone
테스토스테론은 대표적인 동화 호르몬 (Anabolic hormone)으로, 근육대사와 지방대사에 관여한다. 이 호르몬의 수치가 낮아질 경우, 체내 근육량이 감소하여 기초대사량이 낮아지고 비만을 쉽게 야 기하게 된다. 또한 갱년기의 대표적인 증상 중 하나인 피부 각질화를 유도하게 된다. 도 3을 살펴보면, 난소절제를 실시한 모든 군에서 비 난소절제 대조군에 비해 혈중 테스토스테론의 농도가 감소하였다. 이 중 에스트로겐 투여군과 발아 발효콩 1.0 g/kg로 투여한 군에서는 각각 2797.8 ± 517.3 pg/ml, 1969.8 ± 164.2 pg/ml의 테스트스토스테론 농도를 확인할 수 있었고, 이 그룹들의 테스토스테론 농도는 난소절제 대조 군에 비해 유의적으로 (p < 0.05) 증가한 결과임을 확인하였다.Testosterone is a representative anabolic hormone that is involved in muscle and fat metabolism. Lower levels of these hormones reduce muscle mass in the body, which leads to lower basal metabolism and easier obesity. It also induces skin keratinization, one of the typical symptoms of menopause. Referring to FIG. 3, the concentration of testosterone in blood was decreased in all the groups that underwent ovarian ablation compared to the non-ovarian ablation control group. Among them, the testosterone concentrations of 2797.8 ± 517.3 pg / ml and 1969.8 ± 164.2 pg / ml were found in the estrogen-administered group and 1.0 g / kg of germinated fermented soybean, respectively. It was confirmed that the results were significantly increased (p <0.05) compared to the control group.
2.4. 골대사 지표 측정 2.4. Bone metabolism indicator measurement
도 4를 보면, 혈중 칼슘 농도는 그룹 간의 유의적인 차이가 나타나지 않았고, 인의 경우, 발아 발효콩 1.0 g/kg를 투여한 군에서 유의적 차이를 보였다(p < 0.05)). 4, there was no significant difference in blood calcium levels among the groups, and in the case of phosphorus, there was a significant difference in the group administered with 1.0 g / kg of germinated fermented soybeans (p <0.05)).
Alkaline phosphatase(ALP)는 골대사에 관여하는 조 골세포의 biomarker이다. 골다공증과 같이 골대사가 급격히 증가하는 질병이 있을 경우, ALP의 수치 또한 증가하게 된다. 난소절제 대조군의 혈중 ALP 수치는 454.0 ± 69.9 U/L으로 비 난소절제 대조군의 혈중 ALP 수 치 340.2 ± 33.31 U/L 에 비해 증가하였다. 반면, 발아 발효콩 0.5 g/kg를 투여한 군에서 260.0 ± 646.9 U /L로, 대조군에 비해 유의적으로 낮은 ALP 활성을 보였다(p < 0.05). 또한, 난소절제 대조군에 비해 발아발효콩 0.5 g/kg를 투여한 군에서 Bone-ALP의 수치가 140.8 ± 58.5 U/L.로 나타나, 유의적으로 (p < 0.05) 낮 아지는 것을 확인할 수 있었다.Alkaline phosphatase (ALP) is a biomarker of osteoblasts involved in bone metabolism. In other diseases, such as osteoporosis, in which bone metabolism increases rapidly, the level of ALP also increases. The blood ALP level of the ovarian control group was 454.0 ± 69.9 U / L, which was higher than that of the non-ovarian control group (340.2 ± 33.31 U / L). On the other hand, in the group administered 0.5 g / kg of germinated fermented soybeans, 260.0 ± 646.9 U / L, significantly lower ALP activity than the control group (p <0.05). In addition, the Bone-ALP level was 140.8 ± 58.5 U / L. In the group administered with 0.5 g / kg of germinated fermented soybean compared to the control group of ovarian resection, indicating that it was significantly lower (p <0.05). .
도 5를 살펴보면, 6주간의 경구투여를 실시한 뒤, 난소절제 대 조군에서 비 난소절제 대조군와 비교했을 때 유의적으로 (p < 0.05) 골염량이 낮아지는 것을 확인할 수 있었 다. 발아 발효콩 1.0 g/kg, 2.0 g/kg로 투여한 군에서 각각의 골염량은 61.3 ± 0.36%, 60.8 ± 0.31%으로 나 타났다. 이는 난소절제 대조군에 비해 유의적으로 (p < 0.05) 골염량이 증가한 수치이다. 이를 통해 발아 발효콩 투여가 골의 미네랄 함량을 증가시키는 것을 확인할 수 있었다.Referring to FIG. 5, after oral administration for 6 weeks, it was confirmed that the bone inflammation was significantly lower (p <0.05) in the ovarian resection group compared with the non-ovarian resection control group. In the groups administered with 1.0 g / kg and 2.0 g / kg of germinated fermented soybeans, the bone salts were 61.3 ± 0.36% and 60.8 ± 0.31%, respectively. This is a significant (p <0.05) increase in bone salt compared to ovarian resection control. This confirmed that the germination of fermented soybeans increased the mineral content of bone.
2.5. 골조직 형태 측정: Micro-CT 촬영2.5. Bone Morphology Measurement: Micro-CT
난소절제를 한 모든 군에서 비 난소절제 군에 비해 percent bone volume(BV/TV (%)), trabecular number(Tb.N (mm-1)), trabecular thickness(Tb.Th (μm))의 수치가 감소한 것을 확인할 수 있다. 에스트로겐 투여군과 발아 발효콩 2.0 g/kg 투여한 군에서 난소절제 대 조군에 비해 유의적으로 그 수치들이 증가하였다. BV/TV 의 경우, 난소절제 대조군에서 13.7 ± 1.3%로 감소하였으며, 발아 발효콩 2.0 g/kg를 투여한 군의 BT/TV 가 23.3 ± 1.3% 으로 나타나, 유의적인 차이를 보였다 (p < 0.01) (도 6). Tb.N 의 경우, 난소절제 대조군에서 1.17 ± 0.09 mm-1로 낮은 수치를 보였으나, 발아 발효콩 2.0 g/kg를 투여한 군에서 1.69 ± 0.15 mm-1로 에스트로겐 투여군과 유사한 수치로 증가한 것을 확인할 수 있었다(p < 0.05). Tb.Th 의 경우, 난소절제 대조군에서 0.12 ± 0.00 mm-1로 낮은 수치를 보였으나, 에스트로겐 및 발아 발효콩을 투여한 모든 농도에서 Tb.Th 가 유의적으로 증가하였다(도 6). 이는 발아 발효콩을 투여한 쥐에서 골의 밀도가 유의적으로 증가했고 , 해면골의 두께와 수가 증가하였다는 것을 의미한다. The values of percent bone volume (BV / TV (%)), trabecular number (Tb.N (mm -1 )), and trabecular thickness (Tb.Th (μm)) in all ovarian sections compared to non-ovarian sections It can be seen that decreases. The estrogen-treated group and the 2.0 g / kg germinated fermented soybean group increased significantly compared to the ovarian ablation control group. BV / TV was decreased to 13.7 ± 1.3% in the control group of ovarian ablation, and BT / TV of the group treated with 2.0 g / kg of germinated fermented soybean was 23.3 ± 1.3%, showing a significant difference (p <0.01 ) (FIG. 6). In the case of Tb.N, the ovarian control group showed a low value of 1.17 ± 0.09 mm -1 , but increased to 1.69 ± 0.15 mm -1 similar to that of the estrogen group in the 2.0 g / kg germinated fermented soybean group. It could be confirmed (p <0.05). In the case of Tb.Th, the ovarian control group showed a low value of 0.12 ± 0.00 mm −1 , but Tb.Th was significantly increased at all concentrations of estrogen and germinated fermented soybeans (FIG. 6). This means that the bone density of the germinated fermented soybeans increased significantly, and the thickness and number of spongy bones increased.
도 6을 보면, Trabecular separation (Tb .Sp (μm)), trabecular pattern factor (Tb.pf (mm-1))와 structure model index (SMI)의 경우 난소절제를 한 모든 군에서 비 난소절제 대조군에 비해 그 수치들이 증가했다. 발아 발효콩 2.0 g/kg 투여한 군에서 Tb.Sp 수치는 0.70 ± 0.10 으로 나타나 유의적으로 감소하였다(p < 0.05). 이 결과를 통해 난소절제 대조군에 비해 발아 발효콩을 투여한 군에서 골다공증과 밀접한 관련이 있는 해면골의 분산도가 감소했음을 알 수 있다. Tb.pf 의 경우, 에스트로겐 투여군과 발아 발효콩 2.0 g/kg 투여한 군에서 난소절 제 대조군에 비해 유의적으로 (p < 0.01, p < 0.05) 감소했다. 그 수치는 각각 0.76 ± 0.48 mm -1, 0.63 ± 1.25 mm-1 으로 나타났다. 마지막으로, structure model inde x (SMI)의 경우, 에스트로겐 투여군과 발아 발효콩 2.0 g/kg 투여한 군에서 각각 1.54 ± 0.07 , 1.53 ± 0.10 으로, 난소절제 대조군에 비해 유의적으로(p < 0.01) 감소했다. 따라서 발아 발효콩 2.0 g/kg 투여가 에스 트로겐 투여와 비슷한 수준으로 골다공증을 예방효과가 있음으로 예상한다. 6, in the case of trabecular separation (Tb.Sp (μm)), trabecular pattern factor (Tb.pf (mm −1 )) and structure model index (SMI), all groups with ovarian resection were compared to the non-ovarian resection control group. The figures increased. In the 2.0 g / kg group of germinated fermented soybean, the Tb.Sp level was 0.70 ± 0.10, which was significantly decreased (p <0.05). These results show that the degree of dispersal of spongy bone, which is closely related to osteoporosis, was decreased in the group to which the germinated fermented soybeans were administered compared to the ovarian control group. In the case of Tb.pf, estrogen-treated and 2.0 g / kg of germinated fermented soybeans were significantly decreased (p <0.01, p <0.05) compared to the ovarian control group. The figures were 0.76 ± 0.48 mm -1 and 0.63 ± 1.25 mm -1 , respectively. Finally, the structure model inde x (SMI) was 1.54 ± 0.07, 1.53 ± 0.10 in the estrogen-administered group and 2.0 g / kg of germinated fermented soybean, respectively (p <0.01). Decreased. Therefore, the germination fermented soybean 2.0 g / kg administration is expected to be effective in preventing osteoporosis to a level similar to the administration of estrogen.
3. 간 독성 모델의 항산화 효과3. Antioxidant Effect of Liver Toxicity Model
3.1. In vitro 항산화 활성(DPPH, ABTS, Polyphenol)측정 결과 3.1. In vitro antioxidant activity (DPPH, ABTS, Polyphenol) measurement results
하기 표 2를 살펴보면 각 시료의 항산화 활성을 측정한 결과 항상화 활성은 휴바이오에서 제공한 발아 발효콩보다 70% EtOH을 이용 하여 추출한 발아 발효콩의 항산화 활성 및 Polyphenol함량이 더 높게 측정되었다.Referring to Table 2 below, the antioxidant activity of each sample was measured, and the homeostasis activity of the germinated fermented soybeans extracted with 70% EtOH was higher than the germinated fermented soybeans provided by Hubio.
In vitro 항산화활성(DPPH, ABTS, Polyphenol) 측정결과In vitro antioxidant activity (DPPH, ABTS, Polyphenol) measurement results | |||
IC50ValuedIC 50 Valued | ABTS(mg/ml)ABTS (mg / ml) | DPPH(mg/ml)DPPH (mg / ml) | Polyphenol(μg/ml)Polyphenol (μg / ml) |
GFSGFS | 4.2936±0.034.2936 ± 0.03 | 2.4272±0.042.4272 ± 0.04 | 324.09±9.43324.09 ± 9.43 |
70%에탄올 GFS70% Ethanol GFS | 2.522±0.012.522 ± 0.01 | 0.7366±0.190.7366 ± 0.19 | 684.61±8.94684.61 ± 8.94 |
아스콜빈산(IC50 μg/ml)Ascorbic Acid (IC50 μg / ml) | 68.2545±0.9568.2545 ± 0.95 | 30.6857±1.8730.6857 ± 1.87 | -- |
3.2 실험 재료 (Germinated Fermented and Soybeans : GFS)의 분리 및 정제3.2 Isolation and Purification of Experimental Materials (Germinated Fermented and Soybeans: GFS)
도 8을 살펴보면 항산화 활성이 높게 측정된 70% EtOH추출물을 이용하여 Sep-pak을 이용하여 물질 분리 후 항산화 활성을 측정한 결과 Metanol 30%와 70%에서 높은 Polyphenol 함량을 보여주었다. 또한, 하기 표 3을 살펴보면 항산화 활성 측정 방법인 ABTS 결과 Methanol 70%에 가장 높은 항산화 활성을 나타내었다. 하기 표 3은 In vitro 항산화 활성(ABTS IC50value, Polyphenol)측정 결과를 나타낸 표이다.Referring to FIG. 8, 70% EtOH extract with high antioxidant activity was used to measure the antioxidant activity after separation of the material using Sep-pak. As a result, high polyphenol contents were shown at 30% and 70% of metanol. In addition, looking at the following Table 3 showed the highest antioxidant activity in 70% Methanol results of ABTS results of the antioxidant activity measurement method. Table 3 is a table showing the results of in vitro antioxidant activity (ABTS IC50value, Polyphenol) measurement.
70%에탄올 GFS 분획70% Ethanol GFS Fraction | 1One | 22 | 33 | 44 | 55 | 66 |
ABTS(mg/ml)ABTS (mg / ml) | 2.96±0.072.96 ± 0.07 | 1.16±0.011.16 ± 0.01 | 1.09±0.031.09 ± 0.03 | 0.68±0.010.68 ± 0.01 | 5.62±0.075.62 ± 0.07 | 1.09±0.091.09 ± 0.09 |
Polyphenol(μg/ml)Polyphenol (μg / ml) | 383.1396±7.61383.1396 ± 7.61 | 256.7509±1.63256.7509 ± 1.63 | 298.9451±2.65298.9451 ± 2.65 | 261.6921±1.73261.6921 ± 1.73 | 92.8187±1.5592.8187 ± 1.55 | 988.1484±9.67988.1484 ± 9.67 |
(Ascorbic acid(IC50 ug/ml): 2.13±0.06)(Ascorbic acid (IC 50 ug / ml): 2.13 ± 0.06)
3.3. HepG2 cell을 이용한 t-BHP로 인한 손상으로부터의 시료의 보호 활성 측정3.3. Determination of Protective Activity of Samples from Damage Caused by t-BHP Using HepG2 Cells
3.3.1. 세포생존율3.3.1. Cell survival rate
MTT assay는 노란색의 가용성 tetraz olium 염이 살아있는 세포의 미토콘드리아 succina dehydrogenase에 의해 파란색의 불용성 formazan 생성물로 환원되는 원리는 이용한 실험 방법으로 formazan의 흡광도는 540 nm의 파장에서 최대가 된다. The MTT assay is based on the principle that the yellow soluble tetraz olium salt is reduced to the blue insoluble formazan product by mitochondrial succina dehydrogenase in living cells. The absorbance of formazan is maximized at a wavelength of 540 nm.
이 파장에서 측정된 흡광도는 살아있고 대사적으로 왕성한 세포의 농도를 반영하는 것으로 미토콘드리아 활성의 지표로 사용되고 있다. MTT는 세포 독성도 조사로 시험물질에 어느 정도 독성을 나타내는지 미토콘드리아의 활성을 측정하는 실험으로 시험물질이 독성이 있어 미토콘드리아가 파괴되면 흡광도 값이 낮게 나타난다.The absorbance measured at this wavelength reflects the concentration of living and metabolically active cells and is used as an indicator of mitochondrial activity. MTT is a test for measuring the mitochondrial activity to determine how toxic to a test substance by cytotoxicity test. If the test substance is toxic and the mitochondria are destroyed, the absorbance value is low.
도 9A를 보면, HepG2 cell에 발아 발효콩 시료만 처리한 경우에는 시료를 처리하지 않은 세포(100%) 비교하였을 때 발아발효 콩을 처리한 군에서 0.0312(98.52 ± 3.64), 0.0625(97.08 ± 2.93), 0.125(94.44 ± 1.72), 0.25(96.46 ± 2.86), 0.5(96.19 ± 3.03)mg/mL 농도에서 유의적으로 약간의 독성을 나타내었다. 9A shows that when HepG2 cells were treated with only germinated fermented soybean samples, 0.0312 (98.52 ± 3.64), 0.0625 (97.08 ± 2.93) in the group treated with germinated fermented soybeans when compared with the untreated cells (100%). ), 0.125 (94.44 ± 1.72), 0.25 (96.46 ± 2.86), and 0.5 (96.19 ± 3.03) mg / mL concentrations showed significant toxicity.
또한, 도 9B를 보면 70% EtoH 추출물 발아 발효콩을 이용하여 세포에 처리하였을 때의 세포생존율은 시료를 처리하지 않은 군과 차이가 없었다.In addition, as shown in Figure 9B the cell survival rate when the cells were treated with 70% EtoH extract germinated fermented soybean did not differ from the group not treated with the sample.
아울러, 도 10의 A를 보면, HepG2 cell을 이용하여 발아 발효콩과 t-BHP를 처리한 경우에는 아무것도 처리하지 않은 세포와 비교하였을 때, t-BHP만 처리한 세포의 경우에는 약60% 정도 낮게 나왔으며 t-BHP만 처리한 세포와 비교하였을 때 발아 발효콩 시료를 처리하였을 때는 t-BHP만 처리한 세포와 큰 차이를 볼 수 없었다. 하지만, 도 10B를 살펴보면, 70%EtoH 추출물 발아 발효콩을 t-BHP만 처리한 세포(64.67 ± 0.7 8%)와 비교하였을 때는 농도 유의적이지는 않지만 세포의 생존율이 증가하는 것을 확인할 수 있다. 0.0625( 80.11 ± 0.90%), 0.125(82.67 ± 0.84%), 0.25(83.52 ±1.98%), 0.5(82.54 ± 2.58%)mg/mL.In addition, as shown in Figure 10A, when treated with germinated fermented soybeans and t-BHP using HepG2 cells, compared to cells that did not process anything, about 60% of cells treated with t-BHP only When compared with the cells treated with t-BHP alone, the germinated fermented soybean samples were not significantly different from those treated with t-BHP alone. However, referring to FIG. 10B, when the 70% EtoH extract germinated fermented soybean is compared with the cells treated with t-BHP only (64.67 ± 0.7 8%), the concentration of the cells is not significant but it can be seen that the survival rate of the cells increases. 0.0625 (80.11 ± 0.90%), 0.125 (82.67 ± 0.84%), 0.25 (83.52 ± 1.98%), 0.5 (82.54 ± 2.58%) mg / mL.
3.4. HepG2 cell을 이용한 ROS(reactive oxygen species) 측정 3.4. Reactive oxygen species (ROS) measurement using HepG2 cells
DCFH-DA (dichlorofluorescin diacetate)는 세포내의 활성산소 (ROS)와 반응하여 강한 형광물질인 DCF (dichlorofluorescein)로 전환되게 된다. 따라서 DCF에 의한 형광도를 측정함으로써 ROS의 상대적인 량을 추측할 수 있다. Dichlorofluorescin diacetate (DCFH-DA) reacts with free radicals (ROS) in the cell and converts it into a strong fluorescent substance, dichlorofluorescein (DCF). Therefore, the relative amount of ROS can be estimated by measuring the fluorescence by DCF.
70%EtoH 발아 발효콩 추출물에 대한 ROS 효과를 확인하기 위하여 70%EtoH 발아 발효콩 추출물을 세포독성이 없는 농도로 12시간 동안 배양하고 t-BHP 1mM을 1시간 동안 처리하여 산화적 스트레스를 유발하였다. 여기에 25 μM의 DCFH-DA를 60분 동안 넣은 후에 DCF의 형광도를 측정하였다. In order to confirm the ROS effect on 70% EtoH germinated fermented soybean extract, 70% EtoH germinated fermented soybean extract was incubated for 12 hours in a non-cytotoxic concentration and t- BHP 1 mM was treated for 1 hour to induce oxidative stress. . After 25 μM of DCFH-DA was added for 60 minutes, the fluorescence of DCF was measured.
그 결과는 도 11과 같다. 도 11을 살펴보면, Normal(100 ± 1.00%)에 비해 t-BHP 처리에 의해 (267.62 ± 2.68%)로 형광도가 증가 하였고, 70%EtoH 발아 발효콩 추출물 전처리에 의해서 0.5(195.24 ± 1.95%), 0.25mg/mL(195.24 ± 1.95), 0.125mg/mL(240 ± 2.41), 0.0625mg/mL (252.80 ± 2.53)로 가장 높 은 농도부터 농도 유의적으로 억제되어 70% EtoH 발아 발효콩 추출물에 의한 활성산소 감소효과를 확인할 수 있었다.The result is shown in FIG. Referring to Figure 11, compared with Normal (100 ± 1.00%) fluorescence increased by (267.62 ± 2.68%) by t-BHP treatment, 0.5 (195.24 ± 1.95%) by 70% EtoH germinated fermented soybean extract pretreatment , 0.25mg / mL (195.24 ± 1.95), 0.125mg / mL (240 ± 2.41), 0.0625mg / mL (252.80 ± 2.53), significantly inhibited the concentration from the highest concentration to 70% EtoH germinated fermented soybean extract It was confirmed that the active oxygen reduction effect by.
3.5. 동물모델을 이용한 혈액학적 검사3.5. Hematological tests using animal models
발아 발효콩을 투여하고 t-BHP투여한 군의 혈액 생화학적 검사결과는 아래의 표 4에 나타내었다. Total Cholestrol, LDH, Total Bilirubin, ALP, ALT, 측정한 결과 대조군에 비교하였을 때 유의적인 차이(표 4)가 나타났다. LDH는 해당계 효소로서 체내의 각 조직에 널리 분포되어 있으며, 특히 심장, 간장, 신장 및 근육에 존재하는 혈 청중 LDH는 심질환, 간질환 악성종양 및 백혈병 등에서 증가하나 특이성이 낮다. 급성간염에서는 AST, ALT와 동일하게 간세포로부터 빠져나와서 급성간염초기에 증가되며 만성간염 및 간경변증에서는 정상이거나 경도로 증가한다. 간장애를 나타내는 지표 중의 하나인 AST와 ALT에서 유의적인 차이가 나타났다.Blood biochemical test results of the group administered with germinated fermented soybean and administered with t-BHP are shown in Table 4 below. Total Cholestrol, LDH, Total Bilirubin, ALP, ALT, and measurement results showed a significant difference (Table 4) when compared to the control group. LDH is a glycolytic enzyme and widely distributed in each tissue in the body. Especially, LDH in serum, liver, kidney, and muscle increases in heart disease, liver disease, malignancy, and leukemia, but has low specificity. In acute hepatitis, it escapes from hepatocytes like AST and ALT and increases in early acute hepatitis. In chronic hepatitis and cirrhosis, it increases to normal or mild. Significant differences were found in AST and ALT, one of the indicators of liver failure.
ALT와 AST는 간세포 누출효소로 간세포의 괴사와 간 조직의 파괴가 진행됨에 따라 효소가 혈중으로 유리되어 나타나므로 간독성 연구에 활용되고 있다. 도 12는 ALT와 AST활성을 측정한 결과다. AST는 발아 발효콩의 농도가 0.5 g/kg(83.20 ± 2.86 U/L), 1g/kg(81.80 ± 6.72U/L)로 농도 유의적 변화는 없었지만, 시료 처리군에서 t-BHP만 처리한 군(94.25 ± 7.37 U/L)과 비교하였을 때 유의적으로 낮아졌다. ALT는 발아 발효콩 농도가 0.5g/kg(33.71 ± 5.59 U/L), 1g/kg(38.17 ± 4.49 U/L)로 농도 유의적 변화는 없었지만, 시료 처리군에서 t-BHP만 처리한 군(48.25 ± 16.19 U/L)과 비교하였을 때 유의적으로 낮아졌다. ALT and AST are hepatocellular leak enzymes, which are used for hepatotoxicity studies because enzymes are released into the blood as hepatic necrosis and hepatic tissue destruction progress. 12 shows the results of measuring ALT and AST activity. The concentration of AST was 0.5 g / kg (83.20 ± 2.86 U / L) and 1 g / kg (81.80 ± 6.72 U / L), but there was no significant change in the concentration of germinated fermented soybeans. Significantly lower compared to the group (94.25 ± 7.37 U / L). The ALT showed no significant change in the concentration of germinated fermented soybean at 0.5 g / kg (33.71 ± 5.59 U / L) and 1 g / kg (38.17 ± 4.49 U / L), but only the t-BHP was treated in the sample treatment group. Significantly lower compared to (48.25 ± 16.19 U / L).
TreatmentTreatment | TCHO(mg/dl)TCHO (mg / dl) | LDH(U/Liter)LDH (U / Liter) | TBIL(mg/dl)TBIL (mg / dl) |
NormalNormal | 80.00±7.81abc 80.00 ± 7.81 abc | 123.33±21.83b 123.33 ± 21.83 b | 0.48±0.05ab 0.48 ± 0.05 ab |
t-BHP Controlt-BHP Control | 85.25±3.77a 85.25 ± 3.77 a | 227.0±46.66a 227.0 ± 46.66 a | 0.60±0.22a 0.60 ± 0.22 a |
SilymarinSilymarin | 80.8±6.87ab 80.8 ± 6.87 ab | 141.33±24.34b 141.33 ± 24.34 b | 0.54±0.05ab 0.54 ± 0.05 ab |
GFS 0.5g/kg+t-BHPGFS 0.5g / kg + t-BHP | 66.29±9.89bc 66.29 ± 9.89 bc | 127.0±27.38b 127.0 ± 27.38 b | 0.49±0.07bc 0.49 ± 0.07 bc |
GFS 1g/kg+t-BHP |
64.63±7.89c 64.63 ± 7.89 c | 102.20±16.83b 102.20 ± 16.83 b | 0.41±0.04c 0.41 ± 0.04 c |
3.6. 동물모델을 이용한 MDA측정결과3.6. MDA measurement results using animal models
지질 과산화는 생체에서 자연적으로 발생될 수 있는 superoxide anion radical로부터 유래하는 각종의 free radical에 의해 세포막의 다가 불포화지방산이 과산화되는 현상을 지칭한다. 불포화지방산은 과산화 과정을 통해 분해되어 MDA를 생성하므로 이를 지질과산화를 측정하는 지표로 삼는다. 도 13을 보면, 간조직의 경우 발아 발효콩을 투여한 후 t-BHP를 투여한 군은 시료투여농도 0.5g/kg(34.25 ± 5.80), 1g/kg(32.68 ± 12.57) 에서 각 군간의 유의적인 차이는 없었으나 농도가 높아질수록 감소하였으며 t-BHP만 투여한 군(44.66 ± 1.94)과 비교하였을 때 감소하는 경향을 보였다.Lipid peroxidation refers to a phenomenon in which polyunsaturated fatty acids in cell membranes are peroxidated by various free radicals derived from superoxide anion radicals that can occur naturally in a living body. Unsaturated fatty acids are degraded through peroxidation to produce MDA, which is an indicator of lipid peroxidation. 13, in the case of liver tissue, the group administered t- BHP after the administration of germinated fermented soybeans was significantly different between the groups at the sample concentration of 0.5 g / kg (34.25 ± 5.80) and 1 g / kg (32.68 ± 12.57). There was no difference, but it decreased as the concentration increased and decreased compared to the t- BHP-only group (44.66 ± 1.94).
3.7. 동물모델을 이용한 GST 측정결과3.7. GST measurement results using animal models
GST는 신체의 거의 모든 세포에 존재하고 간의 해독작용뿐만 아니라 많은 신진대사에 관여 하는 효소이다. 특히 GST는 전자친화성 기질을 GSH에 결합시키는 반응을 촉매하는 역할을 하게된다. 도 14를 보면, GST측정 결과 발아 발효콩을 투여한 후 t-BHP를 투여한 군 0.5g/kg, 1g/kg, t-BHP만 투여한 군 (25.68 ± 4.28)과 비교하였을 때 시료투여농도 0.5g/kg (33.65 ± 12.97), 1g/kg (48.43 ± 8.90) 에서 시 료농도가 높아질수록 증가하였으나 t-BHP투여한 군과의 유의적인 차이는 1g/kg에서만 나타났다.GST is an enzyme that exists in almost every cell of the body and is involved in many metabolism as well as detoxification of the liver. In particular, GST catalyzes the reaction of binding an electron-affinity substrate to GSH. Referring to FIG. 14, GST measurements germination fermentation after administration of the beans treated group, t -BHP 0.5g / kg, 1g / kg, only one group administered t -BHP (25.68 ± 4.28) Sample administration as compared to the concentration At 0.5 g / kg (33.65 ± 12.97) and 1 g / kg (48.43 ± 8.90), the sample concentration increased with increasing sample concentration, but significant difference with the t-BHP group was only observed at 1 g / kg.
3.8. 동 물모델을 이용한 GSH 측정결과3.8. GSH measurement result using animal model
GSH는 체내에서 해독기능, 면역기능, 항산화 기능등의 핵심적인 역할을 하는 인체에서는 매우 중요한 해독제이며 강력한 항산화제이다. 도 13을 보면, GSH측정 결과 발아 발효콩을 투여한 후 t-BHP를 투여한 군 0.5g/kg, 1g/kg, t-BHP만 투여한 군21.07 ± 8.33)과 비교하였을 때 시료투여농도 0.5g/kg (25.12 ± 2.44), 1g/kg (24.40 ± 2.52) 에서 시료농도가 높아질수록 유의적 증가는 없었다. 또한 t-BHP투여한 군과의 시료를 투여한군 간의 유의적 차이는 보이지 않았지만, 시료를 투여한 군에서 약간의 증가하는 경향을 보여주었다.GSH is a very important antidote and powerful antioxidant in the human body that plays a key role in detoxification, immune function, and antioxidant function in the body. Referring to FIG. 13, GSH measurements germination fermentation after administration of the beans treated group, t -BHP 0.5g / kg, 1g / kg, t -BHP only administered group 21.07 ± 8.33) Sample administration concentration of 0.5 as compared to the There was no significant increase as the sample concentration increased at g / kg (25.12 ± 2.44) and 1 g / kg (24.40 ± 2.52). In addition, there was no significant difference between the groups receiving the sample and the t- BHP group, but there was a slight increase in the group receiving the sample.
3.9. 동물모델을 이용한 SOD 측정결과3.9. SOD measurement results using animal models
SOD는 superoxide anion(O2)를 H2O2로 환원시키는 작용을 하는 효소로서, cytosol과 mitochondria에 존재한다. Cytosol에는 Cu/Zn-SOD만이 존재하나 mitochondria의 matrix에는 Mn-SOD가, 내막과 외막 사이에는 Cu/Zn-SOD가 소량 존재 한다. SOD측정 결과(도 9) 발아 발효콩을 투여한 후 t-BHP를 투여한 군 0.5g/kg, 1g/kg, t-BHP만 투여한 군(23.11 ± 1.71)과 비교하였을 때 시료투여농도 0.5g/kg (27.35 ± 3.32), 1g/kg (25 .67 ± 0.02) 에서 시료농도가 높아질수록 유의적 증가는 없었다. 또한 t-BHP투여한 군과의 시료를 투여한 군간의 유의적 차이가 있다. 다만, 시료를 투여한 군에서 농도의존적인 효과를 보여주지는 않았다.SOD is an enzyme that reduces superoxide anion (O 2 ) to H 2 O 2 , and is present in cytosol and mitochondria. Only Cu / Zn-SOD exists in the cytosol, but Mn-SOD exists in the matrix of mitochondria, and a small amount of Cu / Zn-SOD exists between the inner and outer layers. SOD measurement results (Fig. 9) germination fermentation after administration of the beans when compared with the group (23.11 ± 1.71) of administration of only the group treated with t -BHP 0.5g / kg, 1g / kg, t -BHP administration sample concentration 0.5 There was no significant increase as the sample concentration increased at g / kg (27.35 ± 3.32) and 1 g / kg (25 .67 ± 0.02). There is also a significant difference between the group administered with the sample and the group administered with t- BHP. However, there was no concentration-dependent effect in the group administered the sample.
3.10. 동물모델을 이용한 CAT 측정결과3.10. CAT measurement results using animal model
Catalase는 SOD의 작용이나 O2의 환원으로 생성된 H2O2를 무독성의 H2O로 환원시켜 산소의 독으로부터 생체를 보호하는 효소로서 대부분 peroxisome에 존재하고 mitochondria에도 소량 존재한다. CAT측정 결과(도 10) 발아 발효콩을 투여한 후 t -BHP를 투여한 군 0.5g/kg, 1g/kg, t-BHP만 투여한 군(182.24 ± 19.29)과 비교하였을 때 시료투여농도 0.5g/kg (204.49 ± 41.51), 1g/kg (293.19 ± 61.02) 에서 시료농도가 높아질수록 유의적 증가하였다. 또한 t-BHP투여한 군과의 시료를 투여한 군간의 유의적 차이가 있으며, 시료를 투여한 군에서 농도의존적인 효과를 보여주었다.Catalase is an enzyme that protects the body from oxygen poison by reducing H 2 O 2 produced by the action of SOD or O 2 reduction to non-toxic H 2 O. Mostly, it is present in peroxisome and in small amounts in mitochondria. CAT measurement results (Fig. 10) after germination fermented beans treated with the group treated with t -BHP 0.5g / kg, 1g / kg, t -BHP only one dose group (182.24 ± 19.29) administration sample concentration of 0.5 as compared to the At g / kg (204.49 ± 41.51) and 1 g / kg (293.19 ± 61.02), the sample concentration increased significantly. In addition, there was a significant difference between the group administered with the sample and the t- BHP group, and the group administered with the sample showed a concentration-dependent effect.
<적용예> <Application example>
본 발명은 발아 발효콩의 추출물 이외에 약리학적, 건강기능식품법, 식품위생법상으로 허용되는 부형제(붕해제, 결합제, 활택제), 안정제, 유화제, 첨가물 등으로 이루어지는 군에서 1종 이상 선택된 것을 포함할 수 있다. 또한, 상기 조성물은 경고제, 과립제, 산제, 정제, 또는 캡슐제 등의 경구용으로 만들어 사용될 수 있다. 상기 조성물의 투여량은 해당 법률에서 정하고 있는 기준(용도, 사용목적, 환자의 상태, 나이, 성별, 체중, 처방약, 질병)에 따라 조절해야 한다.The present invention may include one or more selected from the group consisting of excipients (disintegrants, binders, lubricants), stabilizers, emulsifiers, additives, etc., which are acceptable under the pharmacological, health functional food method, food hygiene method, in addition to the extract of germinated fermented soybean. Can be. In addition, the composition may be made and used for oral use such as warnings, granules, powders, tablets, or capsules. The dosage of the composition should be adjusted according to the criteria set forth in the relevant laws (use, purpose of use, patient's condition, age, sex, weight, prescription drug, disease).
<적용예 1>; 제제 <Application Example 1>; Formulation
제제는 발아 발효콩 추출물에 약사법에서 허용하는 부형제 또는 담체를 섞어 제제로서 산제, 정제, 캡슐제, 환, 과립을 제조하였다. The formulation was prepared by mixing the germinated fermented soybean extract with excipients or carriers acceptable in the pharmacological method to prepare powders, tablets, capsules, pills, and granules as preparations.
<산제> <Product>
발아 발효콩 추출물 2 g 및 유당 1 g을 혼합하고 기밀포에 충진하여 산제를 제조하였다. 2 g of germinated fermented soybean extract and 1 g of lactose were mixed and filled into an airtight cloth to prepare a powder.
<정제> <Tablet>
발아 발효콩 추출물 100 ㎎, 옥수수전분 100 ㎎, 유 당 100 ㎎ 및 스테아린산 마그네 2 ㎎을 혼합한 후, 타정하여 정제를 제조하였다. Germinated fermented soybean extract 100 mg, corn starch 100 mg, lactose 100 mg and stearic acid magnesium 2 mg were mixed and then compressed into tablets.
<캡슐제> <Capsules>
발아 발효콩 추출물 100 ㎎, 옥수수전분 100 ㎎, 유 당 100 ㎎, 스테아린산 마그네슘 2 ㎎을 혼합한 후, 젤라틴 캡슐에 충전하여 제조하였다.Germinated fermented soybean extract 100 mg, corn starch 100 mg, lactose 100 mg, magnesium stearate 2 mg was mixed, and then prepared by filling in gelatin capsules.
<환> <Hwan>
발아 발효콩 추출물 1 g, 유당 1.5 g, 글리세린 1 g, 자일리톨 0.5 g을 혼합한 후, 4 g 크기로 환을 제조하였다.After germination of fermented soybean extract 1 g, lactose 1.5 g, glycerin 1 g, and xylitol 0.5 g, a ring was prepared in a size of 4 g.
<과립><Granule>
발아 발효콩 추출물 150 ㎎, 대두추출물 50 ㎎, 포도당 200 ㎎, 전분 600 ㎎을 프리믹스한 후, 30% 에탄올용액 100 ㎎을 첨가하여 60 ℃에서 건조하여 과립을 형성한 후 포에 충진하였다.After germination of fermented soybean extract 150 mg, soy extract 50 mg, glucose 200 mg, and starch 600 mg, 100 mg of 30% ethanol solution was added and dried at 60 ° C. to form granules, which were then filled into fabric.
<적용예 2>; 식품<Application Example 2>; food
식품은 발아 발효콩 추출물에 식품위생법 또는 건강기능식품법에서 허용하는 부형제 또는 담체를 섞어 건강식품 및 음료를 제조하였다.Foods were prepared by mixing the germinated fermented soybean extract with excipients or carriers allowed by the food hygiene or dietary supplement method.
<건강식품> <Health food>
발아 발효콩 추출물 1g, 초산비타민 A 70 ㎍, 비타민 E 1.0 ㎎, 비타민 0.13 ㎎, 비타민 B2 0.15 ㎎, 비타민 B6 0.5 ㎎, 비타민 B12 0.2 ㎍, 비타민 C 10 ㎎, 비오틴 10 ㎍, 니코틴산아미드 1.7 ㎎, 엽산 50 ㎍, 판토텐산 칼슘 0.5 ㎎, 무기질 혼합물 적량, 황산제1철 1.75 ㎎, 산화아연 0.82 ㎎, 탄산마그네슘 25.3 ㎎, 제1인산칼륨 15 ㎎, 제2인산칼슘 55 ㎎, 구연산칼륨 90 ㎎, 탄산칼슘 100 ㎎, 염화마그네슘 24.8 ㎎를 섞어 건강식품을 제조하였다. Germinated fermented soybean extract 1g, Vitamin A 70 μg, Vitamin E 1.0 mg, Vitamin 0.13 mg, Vitamin B2 0.15 mg, Vitamin B6 0.5 mg, Vitamin B12 0.2 μg, Vitamin C 10 mg, Biotin 10 μg, Nicotinamide 1.7 mg, Folic acid 50 ㎍, calcium pantothenate 0.5 mg, mineral mixture appropriate amount, ferrous sulfate 1.75 mg, zinc oxide 0.82 mg, magnesium carbonate 25.3 mg, potassium monophosphate 15 mg, calcium diphosphate 55 mg, potassium citrate 90 mg, carbonate Health food was prepared by mixing 100 mg of calcium and 24.8 mg of magnesium chloride.
상기에서 발아 발효콩 추출물에 비타민 및 미네랄을 혼합한 비율은 제형이나 수요자층에 따라 다양하게 변형할 수 있다. The proportion of the vitamin and minerals mixed with the germinated fermented soybean extract may be variously modified according to the formulation or the consumer layer.
<음료> <Beverage>
발아 발효콩 추출물 1%, 액상과당 0.5%, 올리고당 2%, 설탕 2%, 식염 0.5% 및 잔부의 정제수와 부재료를 배합하여 살균한 후 포장하여 음료를 제조하였다.Germinated fermented soybean extract 1%, liquid fructose 0.5%, oligosaccharide 2%, sugar 2%, salt 0.5% and the balance of the purified water and subsidiary materials were combined and sterilized and then packaged to prepare a beverage.
상기에서 발아 발효콩 추출물에 당류, 식이섬유, 비타민 및 미네랄을 혼합한 비율은 제형이나 수요자층에 따라 다양하게 변형할 수 있다. The ratio of sugars, dietary fiber, vitamins and minerals to the germinated fermented soybean extract may be variously modified depending on the dosage form or consumer layer.
본 발명의 발아 발효콩 추출물은 혈중 에스트로겐과 테스토스테론 농도가 유의적으로 증가하고, 이로 인해 갱년기 증상을 완화시키고, 골의 미네랄 함량을 증가시켜 골의 밀도가 유의적으로 증가하고, 해면골의 두께와 수가 증가한다. 따라서 산업상 이용가능성이 있다.Germinated fermented soybean extract of the present invention significantly increases blood estrogen and testosterone concentrations, thereby alleviating menopausal symptoms, increasing bone mineral content, significantly increasing bone density, and the thickness and number of sponges Increases. Therefore, there is industrial applicability.
Claims (10)
- 쿠메스테롤을 함유하는 발아 발효콩 추출물. Germinated fermented soybean extract containing cumesterol.
- 제1항에 있어서, 상기 추출물은 간세포 보호용인 것을 특징으로 하는 발아 발효콩 추출물. The germinated fermented soybean extract of claim 1, wherein the extract is for protecting hepatocytes.
- 제1항의 발아 발효콩 추출물을 포함하는 간세포 보호용 조성물. A composition for protecting hepatocytes comprising the germinated fermented soybean extract of claim 1.
- 제1항에 있어서, 상기 추출물은 골 밀도 증가용인 것을 특징으로 하는 발아 발효콩 추출물. The germinated fermented soybean extract according to claim 1, wherein the extract is for increasing bone density.
- 제1항의 발아 발효콩 추출물을 포함하는 골 밀도 증가용 조성물. Composition for increasing bone density comprising the germinated fermented soybean extract of claim 1.
- 제3항 또는 제5항에 있어서, 약사법에서 허용하는 부형제를 더 포함하는 약학 조성물인 것을 특징으로 하는 조성물.The composition according to claim 3 or 5, which is a pharmaceutical composition further comprising an excipient acceptable in the pharmacological method.
- 제3항 또는 제5항에 있어서, 건강기능식품법에서 허용하는 부형제를 더 포함하는 건강기능식품 조성물인 것을 특징으로 하는 조성물.The nutraceutical composition according to claim 3 or 5, further comprising an excipient which is allowed by the nutraceutical method.
- 제3항 또는 제5항에 있어서, 식품위생법에서 허용하는 부형제를 더 포함하는 식품 조성물인 것을 특징으로 하는 조성물.The composition according to claim 3 or 5, which is a food composition further comprising an excipient acceptable in the food hygiene method.
- 제3항 내지 제5항 중 어느 한 항에 있어서, 상기 조성물의 제형은 액제, 과립제, 환제 또는 캡슐인 것을 특징으로 하는 조성물.The composition of any one of claims 3 to 5, wherein the formulation of the composition is a liquid, granule, pill or capsule.
- 대두를 발아시켜 황국균을 접종하여 발효시키고, 여과한 후 건조하여 발아 발효콩을 얻는 단계;Germinating soybeans to inoculate and ferment the Rheum bacteria, and filtering and drying to obtain germinated fermented soybeans;상기 건조된 발아 발효콩에 에탄올 용액을 가하여 분해한 후, 초음파로 열수 추출하여 열수 추출물을 얻는 단계; 및Decomposing by adding ethanol solution to the dried germinated fermented soybeans, and extracting hot water by ultrasonication to obtain a hot water extract; And상기 열수 추출물을 원심분리하여 얻은 발아 발효콩 추출물을 여과한 후, 건조하여 발아 발효콩 추출물을 얻는 단계를 포함하는 것을 특징으로 하는 발아 발효콩 추출물의 제조방법.Filtrating the germinated fermented soybean extract obtained by centrifuging the hot water extract, and then drying to obtain a germinated fermented soybean extract.
Priority Applications (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201680076984.9A CN108430486B (en) | 2015-10-30 | 2016-10-31 | Sprouted fermented bean extract containing coumestrol and composition containing the same |
CN202210422042.9A CN114848702A (en) | 2015-10-30 | 2016-10-31 | Sprouted fermented bean extract containing coumestrol and composition containing the same |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
KR1020150152027A KR20160051658A (en) | 2014-10-31 | 2015-10-30 | Composition for Effect of Protecting Liver Cells and Increasing Bone Density Using Germinated Fermented Soybean Extract Containing |
KR10-2015-0152027 | 2015-10-30 |
Publications (2)
Publication Number | Publication Date |
---|---|
WO2017074159A2 true WO2017074159A2 (en) | 2017-05-04 |
WO2017074159A3 WO2017074159A3 (en) | 2017-06-22 |
Family
ID=58643650
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
PCT/KR2016/012399 WO2017074159A2 (en) | 2015-10-30 | 2016-10-31 | Coumestrol-containing germinated fermented soybean extract having hepatocyte protective effect and bone density increase effect and composition containing same |
Country Status (2)
Country | Link |
---|---|
CN (2) | CN108430486B (en) |
WO (1) | WO2017074159A2 (en) |
Family Cites Families (6)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
KR100706279B1 (en) * | 2005-11-11 | 2007-04-12 | (주)아모레퍼시픽 | Medicine and food compositions for improvement of obesity |
CN102803501B (en) * | 2010-01-29 | 2014-07-30 | 株式会社爱茉莉太平洋 | Method for preparing coumestrol and coumestrol prepared by same |
WO2011122872A2 (en) * | 2010-03-31 | 2011-10-06 | (주)아모레퍼시픽 | Composition comprising coumestrol or a bean extract containing coumestrol |
US20130028921A1 (en) * | 2010-03-31 | 2013-01-31 | Amorepacific Corporation | Composition comprising coumestrol or a bean extract containing coumestrol |
KR102200014B1 (en) * | 2013-05-06 | 2021-01-08 | (주)아모레퍼시픽 | Composition for preventing and treating climacteric symptoms comprising extract of bean containing coumestrol |
KR20160051658A (en) * | 2014-10-31 | 2016-05-11 | (주)휴바이오 | Composition for Effect of Protecting Liver Cells and Increasing Bone Density Using Germinated Fermented Soybean Extract Containing |
-
2016
- 2016-10-31 CN CN201680076984.9A patent/CN108430486B/en active Active
- 2016-10-31 CN CN202210422042.9A patent/CN114848702A/en active Pending
- 2016-10-31 WO PCT/KR2016/012399 patent/WO2017074159A2/en active Application Filing
Also Published As
Publication number | Publication date |
---|---|
CN108430486B (en) | 2022-05-10 |
CN114848702A (en) | 2022-08-05 |
CN108430486A (en) | 2018-08-21 |
WO2017074159A3 (en) | 2017-06-22 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
WO2012050396A2 (en) | Pharmaceutical composition including herbal medicine extract for preventing or treating liver cancer | |
JP5486744B2 (en) | A composition for treating hepatitis, comprising extracts of muyuyo flowers, leaves of thorny banci and turmeric roots | |
EP2144621A2 (en) | Guava extract | |
WO2014058142A1 (en) | Pharmaceutical composition containing aster glehni extract as active ingredientfor preventing or treating obesity or metabolic diseases | |
CA2847543C (en) | Annatto extract compositions, including geranyl geraniols and methods of use | |
JP5038310B2 (en) | Pharmaceutical composition for prevention and / or treatment of bone disease, functional food or health food containing the composition, and pharmaceutical preparation comprising the composition as an active ingredient | |
KR102100295B1 (en) | Germinated Fermented Soybean Extract Having Effect of Protecting Liver Cells and Increasing Bone Density Containing Coumesterol, And Composition Containing The Extract | |
WO2012050397A2 (en) | Pharmaceutical composition including herbal medicine extract for preventing or treating kidney cancer | |
KR100703180B1 (en) | A pharmaceutical composition comprising the extract of herb mixture for treating or preventing osteoporosis disease | |
KR102519649B1 (en) | Composition for the prevention or treatment of prostate-related disease comprising Rhodiola sachalinensis root extract containing kaempferol and epicatechin gallate | |
WO2011115416A2 (en) | Sedum sarmentosum fraction for breaking down alcohol and providing hangover relief | |
KR100733764B1 (en) | Composition comprising the cortex extract of albizzia julibrissin or kuraridinol isolated therefrom for preventing or treating hyperlipidemia | |
KR100817600B1 (en) | Pharmaceutical composition containing pueraria radix extract | |
WO2020235828A1 (en) | Composition for inhibiting osteoclasts containing agastache rugosa extract as active ingredient, and use thereof | |
KR100535872B1 (en) | Composition comprising extract of Glycyrrhizae radix or liquiritigenin for the treatment and prevention of diseases caused by heavy metal poisoning | |
WO2020091265A1 (en) | Composition comprising elderberry extract as effective component for preventing, treating, or alleviating male climacteric syndrome | |
WO2017074159A2 (en) | Coumestrol-containing germinated fermented soybean extract having hepatocyte protective effect and bone density increase effect and composition containing same | |
KR101445966B1 (en) | A composition comprising Amomum cardamomum L. extracts having anti-obesity activity | |
KR20060072642A (en) | Herb extract which are effective on improvement of brain function | |
KR101373493B1 (en) | Composition comprising Hizikia fusiformis for preventing and treating obesity or hyperlipidemia and atherosclerotic-vascular diseases | |
WO2018190638A1 (en) | Composition for preventing or treating corneal diseases, containing glycine max extract | |
WO2013062181A1 (en) | Capsosiphon fulvescens glycoprotein extract having liver protection effect and composition containing same | |
Patil et al. | Cordia dichotoma: a comprehensive review of its phytoconstituents and endophytic fungal metabolites and their potential anticancer effects | |
US10265297B2 (en) | Formulation for the prevention and treatment of bone related disorders | |
WO2015137633A1 (en) | Composite extract of black raspberry oil, raspberry oil and mulberry oil |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
121 | Ep: the epo has been informed by wipo that ep was designated in this application |
Ref document number: 16860342 Country of ref document: EP Kind code of ref document: A2 |
|
NENP | Non-entry into the national phase |
Ref country code: DE |
|
122 | Ep: pct application non-entry in european phase |
Ref document number: 16860342 Country of ref document: EP Kind code of ref document: A2 |