KR102144838B1 - Composition for improving, preventing or treating bone diseases comprising Lactobacillus curvatus WiKim38 or culture medium thereof - Google Patents
Composition for improving, preventing or treating bone diseases comprising Lactobacillus curvatus WiKim38 or culture medium thereof Download PDFInfo
- Publication number
- KR102144838B1 KR102144838B1 KR1020190045533A KR20190045533A KR102144838B1 KR 102144838 B1 KR102144838 B1 KR 102144838B1 KR 1020190045533 A KR1020190045533 A KR 1020190045533A KR 20190045533 A KR20190045533 A KR 20190045533A KR 102144838 B1 KR102144838 B1 KR 102144838B1
- Authority
- KR
- South Korea
- Prior art keywords
- wikim38
- strain
- culture medium
- lactobacillus
- composition
- Prior art date
Links
- 239000001963 growth medium Substances 0.000 title claims abstract description 48
- 239000000203 mixture Substances 0.000 title claims abstract description 29
- 241001134659 Lactobacillus curvatus Species 0.000 title claims abstract description 17
- 208000020084 Bone disease Diseases 0.000 title abstract description 25
- 241000186660 Lactobacillus Species 0.000 claims description 43
- 229940039696 lactobacillus Drugs 0.000 claims description 43
- 208000001132 Osteoporosis Diseases 0.000 claims description 24
- 230000036541 health Effects 0.000 claims description 16
- 239000008194 pharmaceutical composition Substances 0.000 claims description 15
- 235000013376 functional food Nutrition 0.000 claims description 12
- 238000011282 treatment Methods 0.000 claims description 10
- 238000012258 culturing Methods 0.000 claims description 9
- 238000000034 method Methods 0.000 claims description 5
- 239000012141 concentrate Substances 0.000 claims description 4
- 239000012228 culture supernatant Substances 0.000 claims description 4
- 239000000047 product Substances 0.000 claims description 4
- 230000006872 improvement Effects 0.000 claims description 3
- 238000011321 prophylaxis Methods 0.000 claims description 2
- 230000002265 prevention Effects 0.000 claims 1
- 210000002997 osteoclast Anatomy 0.000 abstract description 27
- 230000004069 differentiation Effects 0.000 abstract description 19
- 230000000694 effects Effects 0.000 abstract description 17
- 230000002401 inhibitory effect Effects 0.000 abstract description 9
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 abstract description 8
- 201000010099 disease Diseases 0.000 abstract description 6
- 235000021109 kimchi Nutrition 0.000 abstract description 6
- 230000001009 osteoporotic effect Effects 0.000 abstract description 5
- 239000000243 solution Substances 0.000 description 20
- 210000000988 bone and bone Anatomy 0.000 description 13
- 239000002609 medium Substances 0.000 description 11
- 108090000623 proteins and genes Proteins 0.000 description 11
- 102100024230 Dendritic cell-specific transmembrane protein Human genes 0.000 description 10
- 102100034404 Nuclear factor of activated T-cells, cytoplasmic 1 Human genes 0.000 description 10
- 230000014509 gene expression Effects 0.000 description 10
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 10
- 102000004171 Cathepsin K Human genes 0.000 description 9
- 108020004414 DNA Proteins 0.000 description 9
- 101710190014 Dendritic cell-specific transmembrane protein Proteins 0.000 description 9
- 101710151542 Nuclear factor of activated T-cells, cytoplasmic 1 Proteins 0.000 description 9
- 210000004027 cell Anatomy 0.000 description 9
- 108090000625 Cathepsin K Proteins 0.000 description 8
- 238000009806 oophorectomy Methods 0.000 description 8
- 230000002441 reversible effect Effects 0.000 description 8
- 241000699670 Mus sp. Species 0.000 description 7
- 235000013305 food Nutrition 0.000 description 7
- 210000002540 macrophage Anatomy 0.000 description 7
- 239000007758 minimum essential medium Substances 0.000 description 7
- 241000699666 Mus <mouse, genus> Species 0.000 description 6
- 230000001965 increasing effect Effects 0.000 description 6
- 230000001939 inductive effect Effects 0.000 description 6
- 230000002611 ovarian Effects 0.000 description 6
- 230000026731 phosphorylation Effects 0.000 description 6
- 238000006366 phosphorylation reaction Methods 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- 150000003839 salts Chemical class 0.000 description 6
- 102000007651 Macrophage Colony-Stimulating Factor Human genes 0.000 description 5
- 108010046938 Macrophage Colony-Stimulating Factor Proteins 0.000 description 5
- 101000868216 Mus musculus CD40 ligand Proteins 0.000 description 5
- 235000013361 beverage Nutrition 0.000 description 5
- 230000003247 decreasing effect Effects 0.000 description 5
- 239000004310 lactic acid Substances 0.000 description 5
- 235000014655 lactic acid Nutrition 0.000 description 5
- 238000002271 resection Methods 0.000 description 5
- 241000894006 Bacteria Species 0.000 description 4
- 108091003079 Bovine Serum Albumin Proteins 0.000 description 4
- 206010017076 Fracture Diseases 0.000 description 4
- 102000043136 MAP kinase family Human genes 0.000 description 4
- 108091054455 MAP kinase family Proteins 0.000 description 4
- 230000037182 bone density Effects 0.000 description 4
- 238000004113 cell culture Methods 0.000 description 4
- 230000005754 cellular signaling Effects 0.000 description 4
- 238000005516 engineering process Methods 0.000 description 4
- 238000011156 evaluation Methods 0.000 description 4
- 239000012091 fetal bovine serum Substances 0.000 description 4
- 244000005709 gut microbiome Species 0.000 description 4
- 210000001672 ovary Anatomy 0.000 description 4
- 235000013343 vitamin Nutrition 0.000 description 4
- 239000011782 vitamin Substances 0.000 description 4
- 229940088594 vitamin Drugs 0.000 description 4
- 229930003231 vitamin Natural products 0.000 description 4
- 108010057466 NF-kappa B Proteins 0.000 description 3
- 102000003945 NF-kappa B Human genes 0.000 description 3
- 108091023040 Transcription factor Proteins 0.000 description 3
- 102000040945 Transcription factor Human genes 0.000 description 3
- 150000001720 carbohydrates Chemical class 0.000 description 3
- 235000014633 carbohydrates Nutrition 0.000 description 3
- 230000001054 cortical effect Effects 0.000 description 3
- 239000003937 drug carrier Substances 0.000 description 3
- 239000000796 flavoring agent Substances 0.000 description 3
- 235000013355 food flavoring agent Nutrition 0.000 description 3
- 238000009472 formulation Methods 0.000 description 3
- 239000000499 gel Substances 0.000 description 3
- 229910052500 inorganic mineral Inorganic materials 0.000 description 3
- 239000007788 liquid Substances 0.000 description 3
- 244000005700 microbiome Species 0.000 description 3
- 239000011707 mineral Substances 0.000 description 3
- 230000020395 negative regulation of osteoclast differentiation Effects 0.000 description 3
- -1 s Species 0.000 description 3
- 239000007787 solid Substances 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 206010065687 Bone loss Diseases 0.000 description 2
- 238000011740 C57BL/6 mouse Methods 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- 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 2
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 2
- 101000942967 Homo sapiens Leukemia inhibitory factor Proteins 0.000 description 2
- 102100032352 Leukemia inhibitory factor Human genes 0.000 description 2
- 208000030136 Marchiafava-Bignami Disease Diseases 0.000 description 2
- 208000029725 Metabolic bone disease Diseases 0.000 description 2
- 102000008108 Osteoprotegerin Human genes 0.000 description 2
- 108010035042 Osteoprotegerin Proteins 0.000 description 2
- CZMRCDWAGMRECN-UGDNZRGBSA-N Sucrose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 CZMRCDWAGMRECN-UGDNZRGBSA-N 0.000 description 2
- 229930006000 Sucrose Natural products 0.000 description 2
- 102000008889 TNF receptor-associated factor TRAF Human genes 0.000 description 2
- 108050000808 TNF receptor-associated factor TRAF Proteins 0.000 description 2
- 235000010443 alginic acid Nutrition 0.000 description 2
- 229920000615 alginic acid Polymers 0.000 description 2
- 150000001413 amino acids Chemical class 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 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 2
- 210000004979 bone marrow derived macrophage Anatomy 0.000 description 2
- 239000002299 complementary DNA Substances 0.000 description 2
- 235000008504 concentrate Nutrition 0.000 description 2
- 235000009508 confectionery Nutrition 0.000 description 2
- 235000013365 dairy product Nutrition 0.000 description 2
- 238000002474 experimental method Methods 0.000 description 2
- 238000000855 fermentation Methods 0.000 description 2
- 230000004151 fermentation Effects 0.000 description 2
- 230000003176 fibrotic effect Effects 0.000 description 2
- 235000013373 food additive Nutrition 0.000 description 2
- 239000002778 food additive Substances 0.000 description 2
- 235000003599 food sweetener Nutrition 0.000 description 2
- 235000015203 fruit juice Nutrition 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 230000000968 intestinal effect Effects 0.000 description 2
- 210000000936 intestine Anatomy 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
- 230000007246 mechanism Effects 0.000 description 2
- 230000009245 menopause Effects 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 239000006872 mrs medium Substances 0.000 description 2
- XXUPLYBCNPLTIW-UHFFFAOYSA-N octadec-7-ynoic acid Chemical compound CCCCCCCCCCC#CCCCCCC(O)=O XXUPLYBCNPLTIW-UHFFFAOYSA-N 0.000 description 2
- 210000000056 organ Anatomy 0.000 description 2
- 210000000963 osteoblast Anatomy 0.000 description 2
- 208000005368 osteomalacia Diseases 0.000 description 2
- 208000028169 periodontal disease Diseases 0.000 description 2
- 238000002360 preparation method Methods 0.000 description 2
- 239000003755 preservative agent Substances 0.000 description 2
- 108091006084 receptor activators Proteins 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 208000007442 rickets Diseases 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000003381 stabilizer Substances 0.000 description 2
- UCSJYZPVAKXKNQ-HZYVHMACSA-N streptomycin Chemical compound CN[C@H]1[C@H](O)[C@@H](O)[C@H](CO)O[C@H]1O[C@@H]1[C@](C=O)(O)[C@H](C)O[C@H]1O[C@@H]1[C@@H](NC(N)=N)[C@H](O)[C@@H](NC(N)=N)[C@H](O)[C@H]1O UCSJYZPVAKXKNQ-HZYVHMACSA-N 0.000 description 2
- 229960005322 streptomycin Drugs 0.000 description 2
- 239000005720 sucrose Substances 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 239000003765 sweetening agent Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 230000002195 synergetic effect Effects 0.000 description 2
- 235000013311 vegetables Nutrition 0.000 description 2
- 238000001262 western blot Methods 0.000 description 2
- LNAZSHAWQACDHT-XIYTZBAFSA-N (2r,3r,4s,5r,6s)-4,5-dimethoxy-2-(methoxymethyl)-3-[(2s,3r,4s,5r,6r)-3,4,5-trimethoxy-6-(methoxymethyl)oxan-2-yl]oxy-6-[(2r,3r,4s,5r,6r)-4,5,6-trimethoxy-2-(methoxymethyl)oxan-3-yl]oxyoxane Chemical compound CO[C@@H]1[C@@H](OC)[C@H](OC)[C@@H](COC)O[C@H]1O[C@H]1[C@H](OC)[C@@H](OC)[C@H](O[C@H]2[C@@H]([C@@H](OC)[C@H](OC)O[C@@H]2COC)OC)O[C@@H]1COC LNAZSHAWQACDHT-XIYTZBAFSA-N 0.000 description 1
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- GZCWLCBFPRFLKL-UHFFFAOYSA-N 1-prop-2-ynoxypropan-2-ol Chemical compound CC(O)COCC#C GZCWLCBFPRFLKL-UHFFFAOYSA-N 0.000 description 1
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- FHVDTGUDJYJELY-UHFFFAOYSA-N 6-{[2-carboxy-4,5-dihydroxy-6-(phosphanyloxy)oxan-3-yl]oxy}-4,5-dihydroxy-3-phosphanyloxane-2-carboxylic acid Chemical compound O1C(C(O)=O)C(P)C(O)C(O)C1OC1C(C(O)=O)OC(OP)C(O)C1O FHVDTGUDJYJELY-UHFFFAOYSA-N 0.000 description 1
- 244000215068 Acacia senegal Species 0.000 description 1
- 235000006491 Acacia senegal Nutrition 0.000 description 1
- 102000013563 Acid Phosphatase Human genes 0.000 description 1
- 108010051457 Acid Phosphatase Proteins 0.000 description 1
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 description 1
- 108010011485 Aspartame Proteins 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 206010009900 Colitis ulcerative Diseases 0.000 description 1
- 208000011231 Crohn disease Diseases 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- 201000004624 Dermatitis Diseases 0.000 description 1
- 229920001353 Dextrin Polymers 0.000 description 1
- 239000004375 Dextrin Substances 0.000 description 1
- SNRUBQQJIBEYMU-UHFFFAOYSA-N Dodecane Natural products CCCCCCCCCCCC SNRUBQQJIBEYMU-UHFFFAOYSA-N 0.000 description 1
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 208000004262 Food Hypersensitivity 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
- 108010010803 Gelatin Proteins 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- 229920000084 Gum arabic Polymers 0.000 description 1
- 241000282412 Homo Species 0.000 description 1
- 208000011327 Increased bone mineral density Diseases 0.000 description 1
- 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 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 241000124008 Mammalia Species 0.000 description 1
- 229930195725 Mannitol Natural products 0.000 description 1
- 208000001145 Metabolic Syndrome Diseases 0.000 description 1
- 206010027476 Metastases Diseases 0.000 description 1
- 229920000168 Microcrystalline cellulose Polymers 0.000 description 1
- 101000761505 Mus musculus Cathepsin K Proteins 0.000 description 1
- 101000832059 Mus musculus Dendritic cell-specific transmembrane protein Proteins 0.000 description 1
- 101000995095 Mus musculus Nuclear factor of activated T-cells, cytoplasmic 1 Proteins 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 206010030247 Oestrogen deficiency Diseases 0.000 description 1
- 229920002230 Pectic acid Polymers 0.000 description 1
- 108010025832 RANK Ligand Proteins 0.000 description 1
- 102000014128 RANK Ligand Human genes 0.000 description 1
- 238000013381 RNA quantification Methods 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 244000269722 Thea sinensis Species 0.000 description 1
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 1
- 102100040247 Tumor necrosis factor Human genes 0.000 description 1
- 201000006704 Ulcerative Colitis Diseases 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 201000000690 abdominal obesity-metabolic syndrome Diseases 0.000 description 1
- 235000010489 acacia gum Nutrition 0.000 description 1
- 239000000654 additive Substances 0.000 description 1
- 235000013334 alcoholic beverage Nutrition 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 229940072056 alginate Drugs 0.000 description 1
- 239000000783 alginic acid Substances 0.000 description 1
- 229960001126 alginic acid Drugs 0.000 description 1
- 150000004781 alginic acids Chemical class 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 210000004102 animal cell Anatomy 0.000 description 1
- 239000012736 aqueous medium Substances 0.000 description 1
- 239000000605 aspartame Substances 0.000 description 1
- 235000010357 aspartame Nutrition 0.000 description 1
- IAOZJIPTCAWIRG-QWRGUYRKSA-N aspartame Chemical compound OC(=O)C[C@H](N)C(=O)N[C@H](C(=O)OC)CC1=CC=CC=C1 IAOZJIPTCAWIRG-QWRGUYRKSA-N 0.000 description 1
- 229960003438 aspartame Drugs 0.000 description 1
- 208000006673 asthma Diseases 0.000 description 1
- 208000010668 atopic eczema Diseases 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000037118 bone strength Effects 0.000 description 1
- 235000008429 bread Nutrition 0.000 description 1
- 238000009395 breeding Methods 0.000 description 1
- 230000001488 breeding effect Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000001506 calcium phosphate Substances 0.000 description 1
- 229910000389 calcium phosphate Inorganic materials 0.000 description 1
- 229960001714 calcium phosphate Drugs 0.000 description 1
- 235000011010 calcium phosphates Nutrition 0.000 description 1
- 239000000378 calcium silicate Substances 0.000 description 1
- 229910052918 calcium silicate Inorganic materials 0.000 description 1
- 229960003340 calcium silicate Drugs 0.000 description 1
- 235000012241 calcium silicate Nutrition 0.000 description 1
- OYACROKNLOSFPA-UHFFFAOYSA-N calcium;dioxido(oxo)silane Chemical compound [Ca+2].[O-][Si]([O-])=O OYACROKNLOSFPA-UHFFFAOYSA-N 0.000 description 1
- 201000011510 cancer Diseases 0.000 description 1
- 239000002775 capsule Substances 0.000 description 1
- 235000014171 carbonated beverage Nutrition 0.000 description 1
- 230000015556 catabolic process Effects 0.000 description 1
- 230000010261 cell growth Effects 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 235000010980 cellulose Nutrition 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 235000019219 chocolate Nutrition 0.000 description 1
- 239000003593 chromogenic compound Substances 0.000 description 1
- 239000003086 colorant Substances 0.000 description 1
- 238000002591 computed tomography Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000000093 cytochemical effect Effects 0.000 description 1
- 238000006731 degradation reaction Methods 0.000 description 1
- 230000001419 dependent effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 235000019425 dextrin Nutrition 0.000 description 1
- 239000008121 dextrose Substances 0.000 description 1
- 206010012601 diabetes mellitus Diseases 0.000 description 1
- 150000002016 disaccharides Chemical class 0.000 description 1
- 230000006806 disease prevention Effects 0.000 description 1
- 239000002270 dispersing agent Substances 0.000 description 1
- 239000002552 dosage form Substances 0.000 description 1
- 239000003792 electrolyte Substances 0.000 description 1
- 239000003995 emulsifying agent Substances 0.000 description 1
- 239000000839 emulsion Substances 0.000 description 1
- 210000003890 endocrine cell Anatomy 0.000 description 1
- 230000007613 environmental effect Effects 0.000 description 1
- 239000003797 essential amino acid Substances 0.000 description 1
- 235000020776 essential amino acid Nutrition 0.000 description 1
- 235000020774 essential nutrients Nutrition 0.000 description 1
- 229940011871 estrogen Drugs 0.000 description 1
- 239000000262 estrogen Substances 0.000 description 1
- 230000029142 excretion Effects 0.000 description 1
- 239000000284 extract Substances 0.000 description 1
- 235000020932 food allergy Nutrition 0.000 description 1
- 235000012041 food component Nutrition 0.000 description 1
- 239000005417 food ingredient Substances 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 229940014259 gelatin Drugs 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 239000008103 glucose Substances 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 235000011187 glycerol Nutrition 0.000 description 1
- 239000008187 granular material Substances 0.000 description 1
- 230000005802 health problem Effects 0.000 description 1
- 239000000017 hydrogel Substances 0.000 description 1
- 235000015243 ice cream Nutrition 0.000 description 1
- 210000000987 immune system Anatomy 0.000 description 1
- 230000006698 induction Effects 0.000 description 1
- 230000004968 inflammatory condition Effects 0.000 description 1
- 230000002757 inflammatory effect Effects 0.000 description 1
- 239000004615 ingredient Substances 0.000 description 1
- 230000003870 intestinal permeability Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 238000007918 intramuscular administration Methods 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000001990 intravenous administration Methods 0.000 description 1
- 239000008101 lactose Substances 0.000 description 1
- 239000003446 ligand Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 235000010355 mannitol Nutrition 0.000 description 1
- 239000000594 mannitol Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000003550 marker Substances 0.000 description 1
- 235000013372 meat Nutrition 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 230000009401 metastasis Effects 0.000 description 1
- 208000037819 metastatic cancer Diseases 0.000 description 1
- 208000011575 metastatic malignant neoplasm Diseases 0.000 description 1
- 229920000609 methyl cellulose Polymers 0.000 description 1
- 239000001923 methylcellulose Substances 0.000 description 1
- 235000010981 methylcellulose Nutrition 0.000 description 1
- LXCFILQKKLGQFO-UHFFFAOYSA-N methylparaben Chemical compound COC(=O)C1=CC=C(O)C=C1 LXCFILQKKLGQFO-UHFFFAOYSA-N 0.000 description 1
- 238000010603 microCT Methods 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 235000019813 microcrystalline cellulose Nutrition 0.000 description 1
- 239000008108 microcrystalline cellulose Substances 0.000 description 1
- 229940016286 microcrystalline cellulose Drugs 0.000 description 1
- 235000010446 mineral oil Nutrition 0.000 description 1
- 239000002480 mineral oil Substances 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000005088 multinucleated cell Anatomy 0.000 description 1
- 210000001721 multinucleated osteoclast Anatomy 0.000 description 1
- 201000006417 multiple sclerosis Diseases 0.000 description 1
- 235000021096 natural sweeteners Nutrition 0.000 description 1
- 210000000653 nervous system Anatomy 0.000 description 1
- 235000012149 noodles Nutrition 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 235000005985 organic acids Nutrition 0.000 description 1
- 238000011633 osteoporosis animal model Methods 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- LCLHHZYHLXDRQG-ZNKJPWOQSA-N pectic acid Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)O[C@H](C(O)=O)[C@@H]1OC1[C@H](O)[C@@H](O)[C@@H](OC2[C@@H]([C@@H](O)[C@@H](O)[C@H](O2)C(O)=O)O)[C@@H](C(O)=O)O1 LCLHHZYHLXDRQG-ZNKJPWOQSA-N 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 235000013550 pizza Nutrition 0.000 description 1
- 229920003023 plastic Polymers 0.000 description 1
- 239000004033 plastic Substances 0.000 description 1
- 239000010318 polygalacturonic acid Substances 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 239000011148 porous material Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002335 preservative effect Effects 0.000 description 1
- 239000006041 probiotic Substances 0.000 description 1
- 235000018291 probiotics Nutrition 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000000069 prophylactic effect Effects 0.000 description 1
- QELSKZZBTMNZEB-UHFFFAOYSA-N propylparaben Chemical compound CCCOC(=O)C1=CC=C(O)C=C1 QELSKZZBTMNZEB-UHFFFAOYSA-N 0.000 description 1
- 229960003415 propylparaben Drugs 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 206010039073 rheumatoid arthritis Diseases 0.000 description 1
- 235000019204 saccharin Nutrition 0.000 description 1
- CVHZOJJKTDOEJC-UHFFFAOYSA-N saccharin Chemical compound C1=CC=C2C(=O)NS(=O)(=O)C2=C1 CVHZOJJKTDOEJC-UHFFFAOYSA-N 0.000 description 1
- 229940081974 saccharin Drugs 0.000 description 1
- 239000000901 saccharin and its Na,K and Ca salt Substances 0.000 description 1
- 235000013580 sausages Nutrition 0.000 description 1
- HFHDHCJBZVLPGP-UHFFFAOYSA-N schardinger α-dextrin Chemical compound O1C(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC(C(O)C2O)C(CO)OC2OC(C(C2O)O)C(CO)OC2OC2C(O)C(O)C1OC2CO HFHDHCJBZVLPGP-UHFFFAOYSA-N 0.000 description 1
- 230000019491 signal transduction Effects 0.000 description 1
- 230000007727 signaling mechanism Effects 0.000 description 1
- 235000011888 snacks Nutrition 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 238000002415 sodium dodecyl sulfate polyacrylamide gel electrophoresis Methods 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 235000010356 sorbitol Nutrition 0.000 description 1
- 235000014347 soups Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 210000000130 stem cell Anatomy 0.000 description 1
- 238000007920 subcutaneous administration Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 239000000725 suspension Substances 0.000 description 1
- 239000006188 syrup Substances 0.000 description 1
- 235000020357 syrup Nutrition 0.000 description 1
- 239000003826 tablet Substances 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 235000012222 talc Nutrition 0.000 description 1
- 235000013616 tea Nutrition 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 239000002562 thickening agent Substances 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- QORWJWZARLRLPR-UHFFFAOYSA-H tricalcium bis(phosphate) Chemical compound [Ca+2].[Ca+2].[Ca+2].[O-]P([O-])([O-])=O.[O-]P([O-])([O-])=O QORWJWZARLRLPR-UHFFFAOYSA-H 0.000 description 1
- 210000004291 uterus Anatomy 0.000 description 1
- 150000003722 vitamin derivatives Chemical class 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 239000000080 wetting agent Substances 0.000 description 1
Images
Classifications
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- 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
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- 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
- A61K9/14—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles
- A61K9/19—Particulate form, e.g. powders, Processes for size reducing of pure drugs or the resulting products, Pure drug nanoparticles lyophilised, i.e. freeze-dried, solutions or dispersions
-
- 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
-
- 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
- A23V2200/306—Foods, ingredients or supplements having a functional effect on health having an effect on bone mass, e.g. osteoporosis prevention
-
- 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
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/133—Curvatus
-
- A23Y2220/25—
-
- C12R1/225—
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Medicinal Chemistry (AREA)
- Organic Chemistry (AREA)
- Mycology (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Epidemiology (AREA)
- Wood Science & Technology (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Zoology (AREA)
- Tropical Medicine & Parasitology (AREA)
- Polymers & Plastics (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Biochemistry (AREA)
- Molecular Biology (AREA)
- Physical Education & Sports Medicine (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
Abstract
Description
본 발명은 락토바실러스 쿠르바투스(Lactobacillus curvatus) WiKim38 균주 또는 이의 배양액을 포함하는 골 질환 개선, 예방 또는 치료용 조성물에 관한 것으로서, 더욱 상세하게는 김치로부터 분리된 락토바실러스 쿠르바투스 WiKim38 균주 또는 이의 배양액을 포함하는 조성물의 골 질환 개선, 예방 또는 치료 효과에 관한 것이다.The present invention relates to a composition for improving, preventing or treating bone diseases, including Lactobacillus curvatus WiKim38 strain or a culture thereof, and more particularly, Lactobacillus curvatus WiKim38 strain isolated from kimchi or its It relates to the effect of improving, preventing or treating bone diseases of a composition comprising a culture medium.
갱년기의 여성에게서는 여러 폐경 증후들이 나타나는데, 특히 에스트로겐의 감소로 인하여 뼈에서 칼슘이 빠져나가 뼈의 질량이 감소하고 구멍이 많아지며 골 손실의 증가 등으로 인하여 골다공증 질환의 발병률이 높아지게 된다. 폐경기 이후 후기변화는 증상이 나타나기까지 다소 시간이 걸릴 수 있으므로 문제를 쉽게 자각하지 못하는 경우도 많다. 보건복지부 국민 건강 통계치에 따르면, 30세 이상에서의 골다공증 유병률이 남성은 1%, 여성은 9%로 여성의 유병률이 남성 대비 9배나 높게 나타난 것으로 보고되었다.In menopausal women, various symptoms of menopause appear.In particular, the incidence of osteoporosis disease increases due to the loss of estrogen, which causes calcium to escape from the bones, resulting in a decrease in bone mass, increased pores, and increased bone loss. Late changes after menopause may take some time for symptoms to appear, so there are many cases where problems are not easily recognized. According to the National Health Statistics of the Ministry of Health and Welfare, the prevalence of osteoporosis in men over the age of 30 was 1% and 9% in women, and it was reported that the prevalence of women was 9 times higher than that of men.
골다공증에 의해 야기되는 골절은 주된 건강 문제를 구성하며 건강 관리 시스템 상에 막대한 경제적 부담을 준다. 골다공증으로 인한 골절의 위험은 서양에서 높으며(여성에서 약 50% 및 남성에서 약 20%), 골절은 현저한 치사율 및 이환율과 연관된다. 피질골(cortical bone)은 신체에서 뼈의 약 80%를 구성하며, 여러 연구들은 피질골이 뼈 강도의 주요 결정인자이며, 이에 따라 골절 민감성의 주요 결정인자임을 보여주었다. 65세 이후에 뼈 손실은 해면질골이 아니라 주로 피질골에서의 손실에 기인한다(Lancet, 2010, May 15; 375(9727):1729-36).Fractures caused by osteoporosis constitute a major health problem and place a huge economic burden on the health care system. The risk of fractures due to osteoporosis is high in the West (about 50% in women and about 20% in men), and fractures are associated with significant mortality and morbidity. Cortical bone makes up about 80% of the bones in the body, and several studies have shown that cortical bone is a major determinant of bone strength and thus a major determinant of fracture sensitivity. Bone loss after age 65 is primarily due to loss in cortical bone, not spongy bone (Lancet, 2010, May 15; 375(9727):1729-36).
골격은 뼈 형성 골아세포(OBs) 및 뼈흡수 파골 세포(OCLs)에 의해 리모델링된다. 대식세포 콜로니 자극인자(MCSF)는 OCLs 전구세포의 증식 및 생존을 증가시킬뿐만 아니라 OCL에서 핵인자-κB(RANK)의 리셉터 활성제의 발현을 상향조절한다. 이는 RANK 리간드(RANKL)가 바인딩하고 OCL 형성을 이끄는 시그널링 캐스케이드를 개시하도록 한다. RANKL의 영향은 RANKL에 대한 유인 리셉터인 오스테오프로테게린(OPG)에 의해 저해될 수 있다.The skeleton is remodeled by bone-forming osteoblasts (OBs) and bone resorptive osteoclasts (OCLs). Macrophage colony stimulating factor (MCSF) not only increases proliferation and survival of OCLs progenitor cells, but also upregulates the expression of receptor activators of nuclear factor-κB (RANK) in OCL. This allows the RANK ligand (RANKL) to bind and initiate the signaling cascade leading to OCL formation. The effect of RANKL can be inhibited by osteoprotegerin (OPG), an attractive receptor for RANKL.
최근에, 건강 및 질병 모두를 위해 장내 미생물(gut microbiota; 이하 GM)의 중요성이 집중적으로 연구되어 왔다. GM은 총괄적으로 인간 게놈 보다 150배 더 많은 유전자를 함유하는 엄청난 양의 박테리아로 구성된다. 이는 출생시 얻어지며, 구별되는 실재(distinct entity)임에도 불구하고, 분명히 인간 게놈과 함께 공진화하고, 다양한 방식으로 이의 숙주와 소통하고 영향을 미치는 다세포 생물로 간주될 수 있다.Recently, the importance of the gut microbiota (GM) for both health and disease has been intensively studied. GM is collectively made up of a huge amount of bacteria that contain 150 times more genes than the human genome. It is obtained at birth and, despite being a distinct entity, can clearly be considered a multicellular organism that co-evolves with the human genome and communicates and influences its host in a variety of ways.
GM의 구성은 음식 및 항생제 치료와 같은 다수의 환경적 요인에 의해 조절된다. 장내 미생물에 의해 생산된 분자들은 유익할 수도 있고 해로울 수도 있으며, 장 내의 내분비 세포, 장 신경계, 장 투과성 및 면역 시스템에 영향을 미치는 것으로 알려져 있다. 동요된 미생물 구성은 크론병, 궤양성 대장염, 류마티스성 관절염, 다발성 경화증, 당뇨, 음식 알레르기, 습진 및 천식뿐만 아니라 비만 및 대사 증후군을 포함하여 장 내외에서 다양한 염증 조건에 연루되는 것으로 상정되어 왔다.The composition of GM is regulated by a number of environmental factors, such as food and antibiotic therapy. Molecules produced by intestinal microbes can be beneficial or harmful, and are known to affect the endocrine cells in the intestine, the intestinal nervous system, intestinal permeability and the immune system. The disturbed microbial composition has been hypothesized to be implicated in various inflammatory conditions both inside and outside the intestine, including Crohn's disease, ulcerative colitis, rheumatoid arthritis, multiple sclerosis, diabetes, food allergies, eczema and asthma, as well as obesity and metabolic syndrome.
한편, 락토바실러스(Lactobacillus) 속 미생물은 동형 또는 이형발효를 하는 유산간균으로서, 유제품이나 채소의 발효과정에서 흔히 볼 수 있는 균이다. 최근 락토바실러스 속 미생물을 생균제, 식품 첨가제 등으로 개발하고자 하는 연구가 활발히 진행되고 있다.On the other hand, Lactobacillus (Lactobacillus) genus microorganisms are lactic acid bacillus that undergo homogeneous or heterogeneous fermentation, and are commonly found in the fermentation process of dairy products or vegetables. Recently, studies to develop microorganisms in Lactobacillus as probiotics and food additives have been actively conducted.
이에 본 발명자들은 락토바실러스 쿠르바투스(Lactobacillus curvatus) 미생물에 대한 연구를 계속한 결과, 김치로부터 분리된 락토바실러스 쿠르바투스 균주와 그의 배양액이 파골세포 분화 억제와 골다공증 질환 개선 효과를 나타냄을 확인하였다.Accordingly, the present inventors continued research on the Lactobacillus curvatus microorganism, and as a result, it was confirmed that the Lactobacillus curvatus strain isolated from kimchi and its culture medium showed an effect of inhibiting osteoclast differentiation and improving osteoporosis disease. .
이에, 본 발명의 목적은 골 질환 개선, 예방 또는 치료 활성을 가지는 수탁번호 KFCC11667P로 기탁된 락토바실러스 쿠르바투스 WiKim38 균주를 제공하는 것이다.Accordingly, it is an object of the present invention to provide a Lactobacillus curbatus WiKim38 strain deposited with accession number KFCC11667P having an activity to improve, prevent or treat bone diseases.
본 발명의 다른 목적은 골 질환 개선, 예방 또는 치료 활성을 가지는 수탁번호 KFCC11667P로 기탁된 락토바실러스 쿠르바투스 WiKim38 균주의 배양액을 제공하는 것이다.Another object of the present invention is to provide a culture solution of the Lactobacillus curbatus WiKim38 strain deposited under accession number KFCC11667P having an activity to improve, prevent or treat bone disease.
본 발명의 또 다른 목적은 수탁번호 KFCC11667P로 기탁된 락토바실러스 쿠르바투스 WiKim38 균주 또는 이의 배양액을 포함하는 골 질환 예방 또는 치료용 약학적 조성물을 제공하는 것이다.Another object of the present invention is to provide a pharmaceutical composition for preventing or treating bone diseases comprising the Lactobacillus kurbatus WiKim38 strain deposited with accession number KFCC11667P or a culture solution thereof.
본 발명의 또 다른 목적은 수탁번호 KCTC13816BP로 기탁된 락토바실러스 쿠르바투스 WiKim38 균주 또는 이의 배양액을 포함하는 골 질환 개선용 건강기능식품 조성물을 제공하는 것이다.Another object of the present invention is to provide a health functional food composition for improving bone diseases comprising the Lactobacillus kurbatus WiKim38 strain deposited with accession number KCTC13816BP or a culture solution thereof.
본 발명의 또 다른 목적은 수탁번호 KCTC13816BP로 기탁된 락토바실러스 쿠르바투스 WiKim38 균주의 골 질환 개선, 예방 및 치료 용도에 관한 것이다.Another object of the present invention relates to the use of the Lactobacillus curbatus WiKim38 strain deposited under accession number KCTC13816BP to improve, prevent and treat bone diseases.
본 발명은 락토바실러스 쿠르바투스(Lactobacillus curvatus) WiKim38 균주 또는 이의 배양액을 포함하는 골 질환 개선, 예방 또는 치료용 조성물에 관한 것으로, 본 발명에 따른 배양액은 파골세포 분화 억제 효과를 통해 골 질환 개선, 예방 또는 치료 효과를 나타낸다.The present invention relates to a composition for improving, preventing or treating bone diseases, including the Lactobacillus curvatus WiKim38 strain or a culture medium thereof, and the culture medium according to the present invention improves bone disease through an osteoclast differentiation inhibitory effect, It has a prophylactic or therapeutic effect.
본 발명자들은 김치로부터 분리된 락토바실러스 쿠르바투스 WiKim38 균주 또는 이의 배양액이 파골세포의 분화를 억제시키고 골 질환을 치료하는 효과가 있음을 확인하였다.The present inventors have confirmed that the Lactobacillus curbatus WiKim38 strain isolated from kimchi or a culture medium thereof inhibits the differentiation of osteoclasts and has an effect of treating bone diseases.
이하 본 발명을 더욱 자세히 설명하고자 한다.Hereinafter, the present invention will be described in more detail.
본 발명의 일 양태는 골 질환 개선, 예방 또는 치료 활성을 가지는 수탁번호 KFCC11667P로 기탁된 락토바실러스 쿠르바투스 WiKim38 균주이다. 상기 균주는 김치로부터 분리된 것일 수 있으나, 이에 한정되는 것은 아니다.One aspect of the present invention is a Lactobacillus curbatus WiKim38 strain deposited under accession number KFCC11667P having an activity to improve, prevent or treat bone diseases. The strain may be isolated from kimchi, but is not limited thereto.
본 발명의 다른 양태는 골 질환 개선, 예방 또는 치료 활성을 가지는 수탁번호 KFCC11667P로 기탁된 락토바실러스 쿠르바투스 WiKim38 균주의 배양액이다.Another aspect of the present invention is a culture medium of Lactobacillus curbatus WiKim38 strain deposited under accession number KFCC11667P having an activity to improve, prevent or treat bone disease.
본 발명의 다른 양태는 수탁번호 KFCC11667P로 기탁된 락토바실러스 쿠르바투스 WiKim38 균주 또는 이의 배양액을 포함하는 골 질환 예방 또는 치료용 약학적 조성물이다.Another aspect of the present invention is a pharmaceutical composition for preventing or treating bone diseases comprising the Lactobacillus kurbatus WiKim38 strain deposited with accession number KFCC11667P or a culture solution thereof.
상기 골 질환은 골다공증(osteoporosis), 뼈전이암(bone metastatic cancer), 골연화증(osteomalacia), 구루병, 섬유성 골염, 무형성 골질환, 대사성 골질환 및 치주질환으로 이루어진 군으로부터 선택되는 것일 수 있고, 예를 들어, 골다공증일 수 있으나, 이에 한정되는 것은 아니다.The bone disease may be selected from the group consisting of osteoporosis, bone metastatic cancer, osteomalacia, rickets, fibrotic osteopathy, amorphous bone disease, metabolic bone disease, and periodontal disease. For example, it may be osteoporosis, but is not limited thereto.
상기 배양액은, 락토바실러스 쿠르바투스 WiKim38 균주를 배양하고, 상기 균주를 제거한 후 수득된 배양 상층액, 이의 농축물, 이의 분획물 또는 이의 동결건조물인 것일 수 있다.The culture medium may be a culture supernatant obtained after culturing the Lactobacillus curbatus WiKim38 strain and removing the strain, a concentrate thereof, a fraction thereof, or a lyophilized product thereof.
상기 배양액은 락토바실러스 쿠르바투스 WiKim38 균주를 12시간 내지 30시간, 12시간 내지 24시간, 18시간 내지 30시간 또는 18시간 내지 24시간, 예를 들어, 22시간 내지 26시간 동안 배양하여 수득한 것일 수 있으나, 이에 한정되는 것은 아니다. 배양 시간은 24시간을 기점으로 하여 그 이상 배양하는 것이 배양 시간 의존적으로 파골세포 분화 억제 활성에 대하여 현저한 상승효과를 나타내는 것은 아니다.The culture medium was obtained by culturing the Lactobacillus curbatus WiKim38 strain for 12 to 30 hours, 12 to 24 hours, 18 to 30 hours or 18 to 24 hours, for example, 22 to 26 hours. However, it is not limited thereto. The culture time is 24 hours as the starting point, and the culture time does not show a remarkable synergistic effect on the osteoclast differentiation inhibitory activity depending on the culture time.
상기 락토바실러스 쿠르바투스 WiKim38 균주의 배양을 위해서는 Alpha-MEM 배지를 이용할 수 있다.Alpha-MEM medium may be used for culturing the Lactobacillus curbatus WiKim38 strain.
구체적으로, BME(Basal Media Eagle) 배지는 세포성장에 필요한 필수 영양분 및 무기염류와 비타민류에 대한 연구의 결과로 얻어진 배지로 현재 널리 사용되는 MEM과 DMEM의 기본조성이 된 배지이며, MEM(Minimum Essential Media Eagle) 배지는 BME에 비해서 아미노산의 농도가 더 높고 CO2/NaHCO3의 완충 작용을 위해 Earle's salts를 포함하고 있다. 세포 종류에 따라서 비필수아미노산이 더 첨가된 배지와 Earle's salts 대신 Hank's salts가 포함된 배지를 사용하기도 한다. Alpha-MEM(MEM,Alpha modificiation)은 아미노산, 비타민 등이 추가로 첨가되어 있어 동물세포의 DNA 형질전환에 유용하다.Specifically, BME (Basal Media Eagle) medium is a medium obtained as a result of research on essential nutrients, inorganic salts and vitamins necessary for cell growth, and is a medium that is the basic composition of MEM and DMEM, which are widely used today, and MEM (Minimum Essential Media Eagle) has a higher amino acid concentration than BME and contains Earle's salts to buffer CO 2 /NaHCO 3 . Depending on the cell type, a medium containing additional non-essential amino acids and a medium containing Hank's salts instead of Earle's salts may be used. Alpha-MEM (MEM, Alpha modificiation) is useful for DNA transformation of animal cells because amino acids and vitamins are additionally added.
상기 조성물은 배양액을 50 내지 400, 50 내지 350, 50 내지 300, 50 내지 250, 100 내지 400, 100 내지 350, 100 내지 300, 100 내지 250, 150 내지 400, 150 내지 350, 150 내지 300, 150 내지 250, 180 내지 400, 180 내지 350 또는 180 내지 300 ul/ml, 예를 들어, 180 내지 250 ul/ml의 농도로 포함하는 것일 수 있으나, 이에 한정되는 것은 아니다.The composition is a
상기 배양액은 pH 6.5 내지 8.5, 6.5 내지 8.0, 6.5 내지 7.5, 7.0 내지 8.5 또는 7.0 내지 8.0, 예를 들어, pH 7.0 내지 7.5인 것일 수 있으나, 이에 한정되는 것은 아니다.The culture medium may be pH 6.5 to 8.5, 6.5 to 8.0, 6.5 to 7.5, 7.0 to 8.5, or 7.0 to 8.0, for example, pH 7.0 to 7.5, but is not limited thereto.
배양액에서는 유산균이 자라면서 젖산 등의 대사체를 생산하게 되어 산성화될 수 있고, 이는 세포 배양에 적절하지 않은 pH 범위에 해당한다. 이러한 점에 근거하여, 세포 배양에 있어서 가장 적절한 pH 값은 7.4이다.In the culture medium, lactic acid bacteria grow and produce metabolites such as lactic acid, which may be acidified, which corresponds to a pH range that is not suitable for cell culture. Based on this point, the most appropriate pH value for cell culture is 7.4.
상기 골 질환 예방 및 치료는 파골세포 분화 억제를 통해 달성되는 것일 수 있으나, 이에 한정되는 것은 아니다.The bone disease prevention and treatment may be achieved through inhibition of osteoclast differentiation, but is not limited thereto.
본 발명의 약학적 조성물은 락토바실러스 쿠르바투스 WiKim38 균주 배양액의 약학적 유효량 및/또는 약학적으로 허용되는 담체를 포함하는 약학적 조성물로 이용될 수 있다.The pharmaceutical composition of the present invention can be used as a pharmaceutical composition comprising a pharmaceutically effective amount of a culture solution of Lactobacillus curbatus WiKim38 strain and/or a pharmaceutically acceptable carrier.
본 명세서에서 용어 "약학적 유효량"은 상술한 락토바실러스 쿠르바투스 WiKim38 균주 배양액의 효능 또는 활성을 달성하는 데 충분한 양을 의미한다.As used herein, the term "pharmaceutically effective amount" means an amount sufficient to achieve the efficacy or activity of the above-described Lactobacillus curbatus WiKim38 strain culture.
본 발명의 약학적 조성물에 포함되는 약학적으로 허용되는 담체는 제제시에 통상적으로 이용되는 것으로서, 락토스, 덱스트로스, 수크로스, 솔비톨, 만니톨, 전분, 아카시아 고무, 인산 칼슘, 알기네이트, 젤라틴, 규산칼슘, 미세결정성 셀룰로스, 폴리비닐피롤리돈, 셀룰로스, 물, 시럽, 메틸 셀룰로스, 메틸히드록시벤조에이트, 프로필히드록시벤조에이트, 활석, 스테아르산 마그네슘 및 미네랄 오일 등을 포함하나, 이에 한정되는 것은 아니다. 본 발명의 약학적 조성물은 상기 성분들 이외에 윤활제, 습윤제, 감미제, 향미제, 유화제, 현탁제, 보존제 등을 추가로 포함할 수 있다.Pharmaceutically acceptable carriers included in the pharmaceutical composition of the present invention are commonly used at the time of formulation, and include lactose, dextrose, sucrose, sorbitol, mannitol, starch, gum acacia, calcium phosphate, alginate, gelatin, Calcium silicate, microcrystalline cellulose, polyvinylpyrrolidone, cellulose, water, syrup, methyl cellulose, methylhydroxybenzoate, propylhydroxybenzoate, talc, magnesium stearate, mineral oil, etc. It does not become. The pharmaceutical composition of the present invention may further include a lubricant, a wetting agent, a sweetening agent, a flavoring agent, an emulsifying agent, a suspending agent, a preservative, and the like in addition to the above components.
본 발명에 따른 약학적 조성물은 인간을 포함하는 포유동물에 다양한 경로로 투여될 수 있다. 투여 방식은 통상적으로 사용되는 모든 방식일 수 있으며, 예컨대, 경구, 피부, 정맥, 근육, 피하 등의 경로로 투여될 수 있으며, 바람직하게는 경구로 투여될 수 있다.The pharmaceutical composition according to the present invention can be administered to mammals including humans by various routes. The mode of administration may be any method commonly used, for example, may be administered by oral, skin, intravenous, intramuscular, subcutaneous, and the like, and preferably may be administered orally.
본 발명의 약학적 조성물의 적합한 투여량은 제제화 방법, 투여방식, 환자의 연령, 체중, 성별, 병적 상태, 음식, 투여 시간, 투여 경로, 배설 속도 및 반응 감응성과 같은 요인들에 의해 다양하며, 보통으로 숙련된 의사는 소망하는 치료 또는 예방에 효과적인 투여량을 용이하게 결정 및 처방할 수 있다. 본 발명의 바람직한 구현예에 따르면, 본 발명의 약학적 조성물의 1일 투여량은 100 내지 400 mg/kg이다.A suitable dosage of the pharmaceutical composition of the present invention varies depending on factors such as formulation method, mode of administration, age, weight, sex, pathological condition, food, administration time, route of administration, excretion rate and response sensitivity of the patient, Usually the skilled practitioner can readily determine and prescribe the dosage effective for the desired treatment or prophylaxis. According to a preferred embodiment of the present invention, the daily dosage of the pharmaceutical composition of the present invention is 100 to 400 mg/kg.
본 발명의 약학적 조성물은 당해 발명이 속하는 기술 분야에서 통상의 지식을 가진 자가 용이하게 실시할 수 있는 방법에 따라, 약학적으로 허용되는 담체 및/또는 부형제를 이용하여 제제화함으로써 단위 용량 형태로 제조되거나 또는 다용량 용기내에 내입시켜 제조될 수 있다. 이때 제형은 오일 또는 수성 매질중의 용액, 현탁액 또는 유화액 형태이거나 엑스제, 분말제, 과립제, 정제, 캅셀제 또는 젤(예컨대, 하이드로젤) 형태일 수도 있으며, 분산제 또는 안정화제를 추가적으로 포함할 수 있다.The pharmaceutical composition of the present invention is prepared in a unit dosage form by formulating using a pharmaceutically acceptable carrier and/or excipient according to a method that can be easily carried out by a person having ordinary knowledge in the technical field to which the present invention belongs. Or it can be prepared by placing it in a multi-dose container. In this case, the formulation may be in the form of a solution, suspension or emulsion in an oil or aqueous medium, or may be in the form of an extract, powder, granule, tablet, capsule or gel (eg, hydrogel), and may additionally include a dispersant or a stabilizer. .
본 발명의 또 다른 양태는 수탁번호 KFCC11667P로 기탁된 락토바실러스 쿠르바투스 WiKim38 균주 또는 이의 배양액을 포함하는 골 질환 개선용 건강기능식품 조성물이다.Another aspect of the present invention is a health functional food composition for improving bone diseases comprising the Lactobacillus kurbatus WiKim38 strain deposited with accession number KFCC11667P or a culture solution thereof.
상기 골 질환은 골다공증, 뼈전이암, 골연화증, 구루병, 섬유성 골염, 무형성 골질환, 대사성 골질환 및 치주질환으로 이루어진 군으로부터 선택되는 것일 수 있고, 예를 들어, 골다공증일 수 있으나, 이에 한정되는 것은 아니다.The bone disease may be selected from the group consisting of osteoporosis, bone metastasis cancer, osteomalacia, rickets, fibrotic osteoitis, amorphous bone disease, metabolic bone disease, and periodontal disease, and may be, for example, osteoporosis, but is limited thereto. It is not.
상기 배양액은, 락토바실러스 쿠르바투스 WiKim38 균주를 배양하고, 상기 균주를 제거한 후 수득된 배양 상층액, 이의 농축물, 이의 분획물 또는 이의 동결건조물인 것일 수 있다.The culture medium may be a culture supernatant obtained after culturing the Lactobacillus curbatus WiKim38 strain and removing the strain, a concentrate thereof, a fraction thereof, or a lyophilized product thereof.
상기 배양액은 락토바실러스 쿠르바투스 WiKim38 균주를 12시간 내지 36시간, 12시간 내지 30시간, 12시간 내지 24시간, 18시간 내지 36시간, 18시간 내지 30시간 또는 18시간 내지 24시간, 예를 들어, 22시간 내지 26시간 동안 배양하여 수득한 것일 수 있으나, 이에 한정되는 것은 아니다. 배양 시간은 24시간을 기점으로 하여 그 이상 배양하는 것이 배양 시간 의존적으로 파골세포 분화 억제 활성에 대하여 현저한 상승효과를 나타내는 것은 아니다.The culture medium is Lactobacillus curbatus WiKim38 strain 12 hours to 36 hours, 12 hours to 30 hours, 12 hours to 24 hours, 18 hours to 36 hours, 18 hours to 30 hours or 18 hours to 24 hours, for example , It may be obtained by culturing for 22 hours to 26 hours, but is not limited thereto. The culture time is 24 hours as the starting point, and the culture time does not show a remarkable synergistic effect on the osteoclast differentiation inhibitory activity depending on the culture time.
상기 조성물은 배양액을 50 내지 400, 50 내지 350, 50 내지 300, 50 내지 250, 100 내지 400, 100 내지 350, 100 내지 300, 100 내지 250, 150 내지 400, 150 내지 350, 150 내지 300, 150 내지 250, 180 내지 400, 180 내지 350 또는 180 내지 300 ul/ml, 예를 들어, 180 내지 250 ul/ml의 농도로 포함하는 것일 수 있으나, 이에 한정되는 것은 아니다.The composition is a
상기 배양액은 pH 6.5 내지 8.5, 6.5 내지 8.0, 6.5 내지 7.5, 7.0 내지 8.5 또는 7.0 내지 8.0, 예를 들어, pH 7.0 내지 7.5인 것일 수 있으나, 이에 한정되는 것은 아니다.The culture medium may be pH 6.5 to 8.5, 6.5 to 8.0, 6.5 to 7.5, 7.0 to 8.5, or 7.0 to 8.0, for example, pH 7.0 to 7.5, but is not limited thereto.
배양액에서는 유산균이 자라면서 젖산 등의 대사체를 생산하게 되어 산성화될 수 있고, 이는 세포 배양에 적절하지 않은 pH 범위에 해당한다. 이러한 점에 근거하여, 세포 배양에 있어서 가장 적절한 pH 값은 7.4이다.In the culture medium, lactic acid bacteria grow and produce metabolites such as lactic acid, which may be acidified, which corresponds to a pH range that is not suitable for cell culture. Based on this point, the most appropriate pH value for cell culture is 7.4.
상기 골 질환 개선은 파골세포 분화 억제를 통해 달성되는 것일 수 있으나, 이에 한정되는 것은 아니다.The bone disease improvement may be achieved through inhibition of osteoclast differentiation, but is not limited thereto.
본 발명의 건강기능식품 조성물을 식품 첨가물로 사용할 경우, 상기 건강기능식품 조성물을 그대로 첨가하거나 다른 식품 또는 식품 성분과 함께 사용될 수 있고, 통상적인 방법에 따라 적절하게 사용될 수 있다. 일반적으로, 식품 또는 음료의 제조 시에 본 발명의 건강기능식품 조성물은 원료에 대하여 15 중량% 이하, 바람직하게는 10 중량% 이하의 양으로 첨가될 수 있다.When the health functional food composition of the present invention is used as a food additive, the health functional food composition may be added as it is or may be used with other foods or food ingredients, and may be appropriately used according to a conventional method. In general, when preparing food or beverage, the health functional food composition of the present invention may be added in an amount of 15% by weight or less, preferably 10% by weight or less based on the raw material.
상기 식품의 종류에는 특별한 제한은 없다. 상기 물질을 첨가할 수 있는 식품의 예로는 육류, 소세지, 빵, 초콜릿, 캔디류, 스넥류, 과자류, 피자, 라면, 기타 면류, 껌류, 아이스크림류를 포함한 낙농제품, 각종 스프, 음료수, 차, 드링크제, 알콜 음료 및 비타민 복합제 등이 있으며, 통상적인 의미에서의 식품을 모두 포함한다.There is no particular limitation on the type of food. Examples of foods to which the above substances can be added include meat, sausage, bread, chocolate, candy, snacks, confectionery, pizza, ramen, other noodles, gum, dairy products including ice cream, various soups, beverages, tea, drinks, Alcoholic beverages and vitamin complexes, and all foods in the usual sense are included.
상기 음료는 여러 가지 향미제 또는 천연 탄수화물 등을 추가 성분으로서 함유할 수 있다. 상술한 천연 탄수화물은 포도당, 과당과 같은 모노사카라이드, 말토스, 슈크로스와 같은 디사카라이드, 및 덱스트린, 사이클로덱스트린과 같은 천연 감미제나, 사카린, 아스파르탐과 같은 합성 감미제 등을 사용할 수 있다. 상기 천연 탄수화물의 비율은 당업자의 선택에 의해 적절하게 결정될 수 있다.The beverage may contain various flavoring agents or natural carbohydrates as additional ingredients. As the natural carbohydrates described above, monosaccharides such as glucose and fructose, disaccharides such as maltose and sucrose, natural sweeteners such as dextrin and cyclodextrin, synthetic sweeteners such as saccharin and aspartame may be used. . The proportion of the natural carbohydrate can be appropriately determined by the choice of a person skilled in the art.
상기 외에 본 발명의 건강기능식품 조성물은 여러 가지 영양제, 비타민, 전해질, 풍미제, 착색제, 펙트산 및 그의 염, 알긴산 및 그의 염, 유기산, 보호성 콜로이드 증점제, pH 조절제, 안정화제, 방부제, 글리세린, 알콜, 탄산 음료에 사용되는 탄산화제 등을 함유할 수 있다. 그 밖에 본 발명의 건강기능식품 조성물은 천연 과일쥬스, 과일쥬스 음료 및 야채 음료의 제조를 위한 과육을 함유할 수 있다. 이러한 성분은 독립적으로 또는 조합하여 사용할 수 있다. 이러한 첨가제의 비율 또한 당업자에 의해 적절히 선택될 수 있다.In addition to the above, the health functional food composition of the present invention includes various nutrients, vitamins, electrolytes, flavoring agents, coloring agents, pectic acid and salts thereof, alginic acid and salts thereof, organic acids, protective colloidal thickeners, pH adjusters, stabilizers, preservatives, glycerin. , Alcohols, carbonated beverages, and the like. In addition, the health functional food composition of the present invention may contain pulp for the manufacture of natural fruit juice, fruit juice beverage and vegetable beverage. These components may be used independently or in combination. The proportion of these additives can also be appropriately selected by those skilled in the art.
본 발명은 락토바실러스 쿠르바투스(Lactobacillus curvatus) WiKim38 균주 또는 이의 배양액을 포함하는 골 질환 개선, 예방 또는 치료용 조성물에 관한 것으로서, 김치로부터 분리된 락토바실러스 쿠르바투스 균주와 그의 배양액이 파골세포 분화 억제와 골다공증 질환 개선 효과를 나타내므로, 이를 효과적으로 골다공증 질환의 개선, 예방 또는 치료에 이용할 수 있다.The present invention relates to a composition for improving, preventing, or treating bone diseases comprising Lactobacillus curvatus WiKim38 strain or a culture solution thereof, wherein the Lactobacillus curvatus strain isolated from kimchi and its culture medium are differentiated from osteoclasts. Since it exhibits an inhibitory and osteoporotic disease improvement effect, it can be effectively used for improving, preventing or treating osteoporotic disease.
도 1a는 락토바실러스 쿠르바투스(Lactobacillus curvatus) WiKim38 균주 배양액의 파골세포 분화 억제 효과를 나타내는 사진이다.
도 1b는 락토바실러스 쿠르바투스 WiKim38 균주 배양액의 파골세포 분화 억제 효과를 나타내는 그래프이다.
도 2는 락토바실러스 쿠르바투스 WiKim38 균주 배양액의 파골세포 분화 유도 단백질의 인산화 억제 효과를 나타내는 웨스턴 블로팅 결과 사진이다.
도 3은 락토바실러스 쿠르바투스 WiKim38 균주 배양액의 파골세포 분화 유도 유전자의 발현 억제 효과를 나타내는 그래프이다.
도 4는 골다공증을 유도한 마우스 모델에서 락토바실러스 쿠르바투스 WiKim38 균주 및 이의 배양액으로 인한 골밀도 증가 효과를 나타낸 그래프이다.Figure 1a is a photograph showing the effect of inhibiting osteoclast differentiation of the culture medium of Lactobacillus curvatus WiKim38 strain.
1B is a graph showing the effect of inhibiting osteoclast differentiation of the culture medium of Lactobacillus curbatus WiKim38 strain.
Figure 2 is a photograph of the results of Western blotting showing the effect of inhibiting the phosphorylation of osteoclast differentiation-inducing proteins of Lactobacillus curbatus WiKim38 strain culture.
3 is a graph showing the effect of inhibiting the expression of osteoclast differentiation inducing genes of the culture medium of Lactobacillus curbatus WiKim38 strain.
4 is a graph showing the effect of increasing bone mineral density due to the Lactobacillus curbatus WiKim38 strain and a culture medium thereof in a mouse model inducing osteoporosis.
이하, 본 발명을 하기의 실시예에 의하여 더욱 상세히 설명한다. 그러나 이들 실시예는 본 발명을 예시하기 위한 것일 뿐이며, 본 발명의 범위가 이들 실시예에 의하여 한정되는 것은 아니다.Hereinafter, the present invention will be described in more detail by the following examples. However, these examples are for illustrative purposes only, and the scope of the present invention is not limited by these examples.
본 명세서 전체에 걸쳐, 특정 물질의 농도를 나타내기 위하여 사용되는 "%"는 별도의 언급이 없는 경우, 고체/고체는 (중량/중량)%, 고체/액체는 (중량/부피)%, 그리고 액체/액체는 (부피/부피)%이다.Throughout this specification, "%" used to indicate the concentration of a specific substance is (weight/weight)% for solids/solids, (weight/volume)% for solids/liquids, and Liquid/liquid is (vol/vol)%.
실시예 1: 락토바실러스 쿠르바투스 WiKiM38 균주 배양액의 파골세포 분화에 대한 억제 평가Example 1: Evaluation of inhibition of osteoclast differentiation of Lactobacillus curbatus WiKiM38 strain culture medium
세포에 처리하기 위한 배양액을 제조하기 위해 알파(alpha)-MEM 배지에 수탁번호 KFCC11667P로 기탁된 락토바실러스 쿠르바투스(Lactobacillus curvatus) WiKim38 균주를 24시간 동안 배양하고 원심분리 후 상층액을 분리하여 pH 7.4로 맞추어 준비하였으며, 이를 실험에 사용하였다. Lactobacillus curvatus ( Lactobacillus curvatus ) WiKim38 strain deposited with accession number KFCC11667P in alpha-MEM medium to prepare a culture solution for cell treatment for 24 hours, centrifugation, and then separate the supernatant to pH It was prepared according to 7.4, and it was used in the experiment.
마우스 대식세포를 12 웰 플레이트(well plate)에 2 x 105 / 웰(well)의 밀도로 24시간 동안 배양하였으며 소태아혈청(fetal bovine serum; 이하 FBS), 1% 페니실린-스트렙토마이신(Penicillin-Streptomycin; 이하 PS), 대식세포 콜로니 자극인자(macrophage colony stimulating factor; 이하 M-CSF)가 첨가된 배지로 교체해준 후 준비해 둔 배양액 50, 100 및 200 ul/ml을 2시간 동안 처리하였다. 이후 RANKL(Receptor activator of nuclear factor kappa-Β ligand)을 처리하여 24시간 동안 반응시켰다.Mouse macrophages were cultured in a 12-well plate at a density of 2 x 10 5 / well for 24 hours, and fetal bovine serum (FBS), 1% penicillin-streptomycin (Penicillin-) Streptomycin; less PS), macrophage colony stimulating factor (hereinafter referred to as M-CSF) was replaced with a medium added, and the
위와 같은 방법으로 6일 동안 분화시킨 후, 파골세포의 세포 화학적 표지효소인 타타르산염 내성 산성 포스타페이즈(Tartrate resistance acid phosphatase, TRAP)에 발색성 기질을 첨가하여 핵을 염색하였고, 핵이 3개 이상으로 다핵화된 세포를 관찰하고 이미지화하였다.After differentiating for 6 days in the same way as above, the nuclei were stained by adding a chromogenic substrate to tatarate resistance acid phosphatase (TRAP), a cytochemical marker enzyme of osteoclasts. Multinucleated cells were observed and imaged.
대식세포에 RANKL을 처리하면 RANK에 결합하며 TRAP 양성 세포로 분화하게 되는데, 이 TRAP 양성 세포에 RANKL, TNF-α와 같은 염증인자로 자극하면 세포끼리 융합되어 다핵형 TRAP 양성 세포로 분화된다.When the macrophages are treated with RANKL, they bind to RANK and differentiate into TRAP-positive cells. When these TRAP-positive cells are stimulated with inflammatory factors such as RANKL and TNF-α, the cells are fused to each other and differentiate into multinuclear TRAP-positive cells.
도 1a, 1b 및 표 1에서 확인할 수 있듯이, 상기 대식세포에 RANKL을 처리했을 때 파골세포로의 분화가 증가하였고, 배양액을 처리한 경우 농도 의존적으로 파골세포의 수가 현저히 감소하였음을 확인하였다. 이로부터 락토바실러스 쿠르바투스 WiKiM38 균주의 배양액이 파골세포로의 분화를 억제시킴을 확인할 수 있었다.1A, 1B and Table 1, when the macrophages were treated with RANKL, differentiation into osteoclasts increased, and when the culture medium was treated, the number of osteoclasts was significantly decreased in a concentration-dependent manner. From this, it was confirmed that the culture medium of Lactobacillus curbatus WiKiM38 strain inhibited differentiation into osteoclasts.
실시예 2: 락토바실러스 쿠르바투스 WiKiM38 균주 배양액의 파골세포 분화 유도 단백질 발현에 대한 억제 평가Example 2: Evaluation of inhibition of expression of osteoclast differentiation-inducing protein in culture medium of Lactobacillus curbatus WiKiM38 strain
마우스 대식세포에 RANKL 및 락토바실러스 쿠르바투스 WiKim38 균주 배양액을 처리했을 때 MAPK(mitogen-activated protein kinase) 및 NF-κB 신호기전을 확인하고자 하였다. 마우스 대식세포를 12 웰 플레이트에 4 x 105 / 웰의 밀도로 24시간 동안 배양하였다. FBS, 1% PS, M-CSF가 첨가된 배지로 교체해준 후 배양액 100 ul/ml을 2시간 동안 처리하였다.When the mouse macrophages were treated with RANKL and Lactobacillus curbatus WiKim38 strain culture, MAPK (mitogen-activated protein kinase) and NF-κB signaling mechanisms were to be identified. Mouse macrophages were cultured in 12 well plates at a density of 4 x 10 5 /well for 24 hours. After replacing the medium with FBS, 1% PS, and M-CSF added, 100 ul/ml of the culture solution was treated for 2 hours.
이후 RANKL을 처리하여 0, 5, 15 및 30분 동안 반응시켰다. 배지를 제거하고 단백질 용해 버퍼(Protein lysis buffer)를 이용하여 단백질을 추출한 후 정량하여 30 ug의 단백질을 SDS-PAGE 겔(Sodium dodecyl sulfate-Poly arcylamide gel electrophoresis gel)에서 분리하였다. 단백질을 멤브레인(membrane)으로 옮긴 후 1차, 2차 항체(p-jnk(Cell Signaling Technology, #9251S), p-p38(Cell Signaling Technology, 9211S), p-ERK(santa cruz, sc-7383), Ikbalpha(Cell Signaling Technology , 9242S), p-p65(Cell Signaling Technology, 3031S))를 순차적으로 처리하여 키트(ECL solution detection kit, BIO-RAD)로 발현되면 기계(Chemidoc)로 측정하였다.Then, RANKL was treated and reacted for 0, 5, 15 and 30 minutes. After removing the medium and extracting the protein using a protein lysis buffer, 30 ug of protein was separated on an SDS-PAGE gel (Sodium dodecyl sulfate-Poly arcylamide gel electrophoresis gel). After transferring the protein to the membrane, primary and secondary antibodies (p-jnk (Cell Signaling Technology, #9251S), p-p38 (Cell Signaling Technology, 9211S), p-ERK (santa cruz, sc-7383) , Ikbalpha (Cell Signaling Technology, 9242S), p-p65 (Cell Signaling Technology, 3031S)) were sequentially processed and expressed with a kit (ECL solution detection kit, BIO-RAD), and then measured with a machine (Chemidoc).
RANKL은 RANK에 결합하여 TRAF6(TNF receptor associated factor)를 통해서 MAPK 및 NF-κB 단백질 발현을 증가시키고, 여러 전사인자들의 증가를 유도하여 파골세포의 분화를 촉진시킨다. 따라서 이들 기전과 관련하여 락토바실러스 쿠르바투스 WiKim38 균주 배양액이 파골세포 분화를 억제시키는지 확인하기 위하여, MAPK 단백질인 ERK, JNK, p38의 인산화(phosphorylation)를 웨스턴 블로팅(western blotting)을 통해서 확인하였다.RANKL binds to RANK and increases the expression of MAPK and NF-κB proteins through TRAF6 (TNF receptor associated factor), and promotes the differentiation of osteoclasts by inducing an increase in several transcription factors. Therefore, in relation to these mechanisms, in order to confirm whether the culture medium of Lactobacillus curbatus WiKim38 strain inhibits osteoclast differentiation, phosphorylation of MAPK proteins ERK, JNK, p38 was confirmed through western blotting. I did.
도 2에서 확인할 수 있듯이, RANKL 처리에 의하여 5 및 15분째에 MAPK 단백질의 인산화가 증가하였고, 배양액 처리에 의해서 JNK, P38 및 ERK의 인산화가 감소하였음을 확인하였다. 또한 IkB 알파(alpha)의 분해 및 p65의 인산화는 억제되었다. 이로부터 락토바실러스 쿠르바투스 WiKim38 균주 배양액이 JNK, P38 및 ERK의 인산화를 억제함으로써 파골세포로의 분화를 억제하는 것임을 확인하였다.As can be seen in FIG. 2, it was confirmed that phosphorylation of MAPK protein was increased at 5 and 15 minutes by RANKL treatment, and phosphorylation of JNK, P38 and ERK was decreased by treatment with the culture medium. In addition, the degradation of IkB alpha and phosphorylation of p65 were inhibited. From this, it was confirmed that the culture medium of Lactobacillus curbatus WiKim38 strain inhibited the differentiation into osteoclasts by inhibiting phosphorylation of JNK, P38 and ERK.
실시예 3: 락토바실러스 쿠르바투스 WiKiM38 균주 배양액의 파골세포 분화 유도 유전자 발현에 대한 억제 평가Example 3: Evaluation of inhibition of expression of osteoclast differentiation-inducing gene in culture medium of Lactobacillus curbatus WiKiM38 strain
락토바실러스 쿠르바투스 WiKiM38 균주 배양액을 마우스 대식세포에 처리하여 TRAP, DC-STAMP, Cathepsin K, NFATc1에 대한 유전자 발현을 RT(Real-time)-PCR을 통해 확인하였다.Lactobacillus curbatus WiKiM38 strain culture was treated on mouse macrophages, and gene expression for TRAP, DC-STAMP, Cathepsin K, and NFATc1 was confirmed through RT (Real-time)-PCR.
골수-유래 대식세포(bone marrow-derived macrophages; 이하 BMDMs)을 12 웰 플레이트에 1 x 105 / 웰의 밀도로 24시간 동안 배양하였다. 1시간 동안 FBS가 배제된 1% PS만 첨가된 배지를 처리해준 후, 락토바실러스 쿠르바투스 WiKiM38 균주 배양액을 2시간 동안 처리하였다. 이후 RANKL을 처리하여 24시간 동안 반응시켰다. 위와 같은 방법으로 3일간 분화시킨 후 트리졸(TRIzol) 용액을 이용하여 세포 내 RNA를 분리하고 RNA 정량 값을 토대로 RT 프리믹스(premix)를 이용하여 cDNA를 합성하였다. 합성된 cDNA는 프라이머(primer)를 이용하여 RT PCR을 통해 증폭시켰다.Bone marrow-derived macrophages (BMDMs) were cultured in a 12 well plate at a density of 1 x 10 5 /well for 24 hours. After treating the medium to which only 1% PS excluding FBS was added for 1 hour, the culture solution of Lactobacillus curbatus WiKiM38 strain was treated for 2 hours. Then, RANKL was treated and reacted for 24 hours. After differentiation for 3 days in the same manner as above, intracellular RNA was isolated using a trizol solution, and cDNA was synthesized using RT premix based on the RNA quantification value. The synthesized cDNA was amplified through RT PCR using a primer.
사용된 프라이머는 Mouse TRAP, DC-STAMP, Cathepsin K, NFATc1이고 대조군(Control) 유전자인 GAPDH와 비교하여 상대적 발현량을 비교하였다.The primers used were Mouse TRAP, DC-STAMP, Cathepsin K, and NFATc1, and relative expression levels were compared with GAPDH, a control gene.
대식세포에 RANKL을 처리하면 RANK에 결합하여 TRAP 양성인 다핵화된 파골세포로 분화된다. 또한 파골세포로 분화하는데 중요한 전사 인자인 NFATc1을 활성화 시키고 파골세포분화기전(osteoclastogenesis)에 관여하는 TRAP, Cathepsine K, DC-STAMP의 발현을 증가시킨다.When macrophages are treated with RANKL, they bind to RANK and differentiate into TRAP-positive multinucleated osteoclasts. It also activates NFATc1, an important transcription factor for differentiation into osteoclasts, and increases the expression of TRAP, Cathepsine K, and DC-STAMP, which are involved in osteoclastogenesis.
도 3 및 표 3에서 확인할 수 있듯이, RANKL처리에 의하여 증가된 TRAP, DC-STAMP, Cathepsin K, NFATc1의 발현은 락토바실러스 쿠르바투스 WiKiM38 균주 배양액 처리에 의해서 감소하였다. 이로부터 상기 배양액이 전사인자인 NFATc1의 활성을 감소시키고, 파골세포분화기전 관련 유전자인 TRAP, DC-STAMP, Cathepsin K의 발현을 감소시킴으로써, 파골세포로의 분화를 억제시킴을 확인할 수 있었다.As can be seen in Figure 3 and Table 3, the expression of TRAP, DC-STAMP, Cathepsin K, and NFATc1 increased by RANKL treatment was decreased by treatment with Lactobacillus curbatus WiKiM38 strain culture solution. From this, it was confirmed that the culture medium inhibited the differentiation into osteoclasts by reducing the activity of the transcription factor NFATc1 and the expression of the genes related to the osteoclast differentiation mechanism, TRAP, DC-STAMP, and Cathepsin K.
실시예 4: 마우스의 난소적출 수술을 통한 골다공증 동물 모델 유도Example 4: Induction of an animal model of osteoporosis through ovariectomy in mice
난소가 절제된 마우스는 에스트로겐 결핍으로 인해 골밀도가 감소하는 것으로 알려져 골다공증 질환 연구에 많이 이용되고 있으므로, 난소 절제를 통해 마우스에서 골다공증 동물 모델을 유도하였다.Ovarian resected mice are known to have decreased bone density due to estrogen deficiency, and are widely used in osteoporotic disease studies. Thus, an osteoporosis animal model was induced in mice through ovarian resection.
실험동물로는 C57BL/6 암컷(7주령) 마우스를 사용하였다. 22±2℃ 및 상대습도 50±10%, 12시간 명암 주기로 설정된 환경의 플라스틱 케이지에서 사육하였다. 난소절제 이전에 동일 환경에서 약 1주 정도 사육 조건에 적응시켰다.C57BL/6 female (7 weeks old) mice were used as experimental animals. It was reared in a plastic cage in an environment set at 22±2℃ and 50±10% relative humidity, and a 12-hour contrast cycle. Before ovariectomy, they were adapted to breeding conditions for about 1 week in the same environment.
이후 골다공증 유발을 위한 난소적출을 수행하였으며, 졸레틸(zoletil)과 럼펀(lumpun)을 근육 주사하여 마취한 후 난소 수술부위를 제모하고 소독하여 1 cm가량 절개하고, 다른 장기에 손상이 가해지지 않도록 주의하여 자궁을 따라 난소를 확인한 후 봉합용 실로 난소를 결찰한 뒤 양측의 난소를 모두 절제하였다. 난소 절제 후 각 장기를 복강 내로 재위치 시킨 뒤 봉합용 실로 봉합하였다. 난소 절제로부터 10일 이후부터 8주 동안 아래와 같이 치료물질을 투여하였다.After that, ovariectomies were performed to induce osteoporosis, and after intramuscular injection of zoletil and lumpun were anesthetized, the ovarian surgical site was depilated and disinfected, and an incision was made by about 1 cm to prevent damage to other organs. After carefully checking the ovaries along the uterus, the ovaries were ligated with a suture thread, and both ovaries were excised. After ovarian resection, each organ was repositioned into the abdominal cavity and then sutured with a suture thread. Treatment substances were administered as follows for 8 weeks from 10 days after ovarian resection.
균주 투여군 제조: 락토바실러스 쿠르바투스 WiKiM38 균주를 전배양 및 본배양시킨 뒤 1*108 및 1*109 cells/ml로 계산하여 200 ul 경구투여하였다.Preparation of strain administration group: Lactobacillus curbatus WiKiM38 strain was pre-cultured and main-cultured, and then 200 ul was administered orally, calculated as 1*10 8 and 1*10 9 cells/ml.
배양액 투여군 제조: 락토바실러스 쿠르바투스 WiKiM38 균주를 전배양 및 본배양시킨 뒤 1*108 및 1*109 cells/ml 로 계산하였다. 균주는 MRS 배지에 10배 희석하여 24시간 동안 30℃에서 배양하였다. 그 다음 원심분리기를 이용하여 균을 가라앉힌 뒤 배양액인 상층액(sup)을 수집하여 pH 7.4로 맞추어 200 ul 경구투여하였다.Preparation of culture solution administration group: Lactobacillus curbatus WiKiM38 strain was pre-cultured and main cultured, and then 1*10 8 and 1*10 9 cells/ml were calculated. The strain was diluted 10-fold in MRS medium and cultured at 30° C. for 24 hours. Then, after the bacteria were settled using a centrifuge, a supernatant (sup), a culture medium, was collected, adjusted to pH 7.4, and 200 ul orally administered.
실험군은 아래와 같이 설정하였다(C57BL/6 마우스: 58 마리).The experimental group was set as follows (C57BL/6 mice: 58 mice).
① G1: 난소 비절제 대조군(sham), MRS 배지(대조군)투여 (n=6; MRS)① G1: Non-ovarian resection control (sham), MRS medium (control) administration (n=6; MRS)
② G2: 난소 비절제 대조군, 배양액(sup) 원액 투여 (n=6; WiKim38-sup-1)② G2: Ovariectomized control, culture medium (sup) undiluted solution administration (n=6; WiKim38-sup-1)
③ G3: 난소 비절제 대조군, 균주 1*109(cells/ml) 투여 (n=6; WiKim38 109)③ G3: Non-ovarian resection control,
④ G4: 난소 절제 실험군(OVX), MRS 투여 (n=8; MRS)④ G4: Ovariectomy experimental group (OVX), MRS administration (n=8; MRS)
⑤ G5: 난소 절제 실험군, 1/2 희석 배양액 투여 (n=8; WiKim38-sup-0.5)⑤ G5: Ovariectomy experimental group, 1/2 diluted culture solution administration (n=8; WiKim38-sup-0.5)
⑥ G6: 난소 절제 실험군, 배양액 원액 투여 (n=8; WiKim38-sup-1)⑥ G6: Ovariectomy experimental group, culture solution undiluted (n=8; WiKim38-sup-1)
⑦ G7: 난소 절제 실험군, 균주 1*108 투여 (n=8; WiKim38 108)⑦ G7: Ovariectomy experimental group,
⑧ G8: 난소 절제 실험군, 균주 1*109 투여 (n=8; WiKim38 109)⑧ G8: Ovariectomy experimental group,
실시예 5: 락토바실러스 쿠르바투스 유산균 및 그의 배양액 투여에 대한 골다공증 유도 마우스의 골밀도 평가Example 5: Evaluation of Bone Mineral Density in Osteoporosis-induced Mice for Administration of Lactobacillus Courbatus Lactobacillus and Its Culture Solution
골밀도(bone mineral density; BMD) 측정은 이전의 단순 방사선 사진 촬영법이나 컴퓨터 단층 촬영법에 비해 고해상도 영상자료를 제공하는 마이크로 CT(microcomputed tomography)를 이용하였다.Bone mineral density (BMD) was measured using microcomputed tomography (CT), which provides high-resolution image data compared to previous simple radiographs or computed tomography.
실험 종료 후 마우스 다리를 포르말린에 고정시킨 뒤 대구경북첨단의료산업진흥재단 실험동물센터에 골밀도 분석을 의뢰하였다.After the end of the experiment, the mouse leg was fixed in formalin, and the bone density analysis was requested to the Experimental Animal Center of the Daegu Gyeongbuk Advanced Medical Industry Promotion Foundation.
도 4 및 표 4에서 확인할 수 있듯이, 본 연구에서 난소 절제된 마우스 그룹(G4)은 Sham 군들에 비해 유의적으로 골밀도가 감소하는 것으로 관찰되었다. 한편, 락토바실러스 쿠르바투스 균주와 그의 배양액은 골밀도를 증가시키는 것으로 나타났다. 특히 배양액을 1/2 희석한 시료를 투여한 그룹(G5)과 난소 절제된 마우스 그룹(G4)을 비교 시 통계학적으로 유의적인 회복능력을 확인할 수 있었다.As can be seen in Fig. 4 and Table 4, it was observed that in this study, the ovarian resected mouse group (G4) significantly decreased bone density compared to the Sham group. On the other hand, it was found that Lactobacillus curbatus strain and its culture medium increased bone mineral density. In particular, a statistically significant recovery ability was confirmed when comparing the group administered with a sample diluted by 1/2 of the culture medium (G5) and the group of mice whose ovaries were excised (G4).
<110> Industry-University Cooperation Foundation Chonnam University <120> Composition for improving, preventing or treating bone diseases comprising Lactobacillus curvatus WiKim38 or culture medium thereof <130> PN190130 <160> 8 <170> KoPatentIn 3.0 <210> 1 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Mouse TRAP forward primer <400> 1 ctggagtgca cgatgccagc gaca 24 <210> 2 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Mouse TRAP reverse primer <400> 2 tccgtgctcg gcgatggacc aga 23 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> DC-STAMP forward primer <400> 3 ccaaggagtc gtccatgatt 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> DC-STAMP reverse primer <400> 4 ggctgctttg atcgtttctc 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Cathepsin K forward primer <400> 5 ggccaactca agaagaaaac 20 <210> 6 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Cathepsin K reverse primer <400> 6 gtgcttgctt cccttctgg 19 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> NFATc1 forward primer <400> 7 ctcgaaagac agtggagcat 20 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> NFATc1 reverse primer <400> 8 cggctgcctt ccgtctcata g 21 <110> Industry-University Cooperation Foundation Chonnam University <120> Composition for improving, preventing or treating bone diseases comprising Lactobacillus curvatus WiKim38 or culture medium thereof <130> PN190130 <160> 8 <170> KoPatentIn 3.0 <210> 1 <211> 24 <212> DNA <213> Artificial Sequence <220> <223> Mouse TRAP forward primer <400> 1 ctggagtgca cgatgccagc gaca 24 <210> 2 <211> 23 <212> DNA <213> Artificial Sequence <220> <223> Mouse TRAP reverse primer <400> 2 tccgtgctcg gcgatggacc aga 23 <210> 3 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> DC-STAMP forward primer <400> 3 ccaaggagtc gtccatgatt 20 <210> 4 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> DC-STAMP reverse primer <400> 4 ggctgctttg atcgtttctc 20 <210> 5 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> Cathepsin K forward primer <400> 5 ggccaactca agaagaaaac 20 <210> 6 <211> 19 <212> DNA <213> Artificial Sequence <220> <223> Cathepsin K reverse primer <400> 6 gtgcttgctt cccttctgg 19 <210> 7 <211> 20 <212> DNA <213> Artificial Sequence <220> <223> NFATc1 forward primer <400> 7 ctcgaaagac agtggagcat 20 <210> 8 <211> 21 <212> DNA <213> Artificial Sequence <220> <223> NFATc1 reverse primer <400> 8 cggctgcctt ccgtctcata g 21
Claims (14)
The health functional food composition for improving osteoporosis according to claim 9, wherein the culture medium is pH 6.5 to 8.5.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020190045533A KR102144838B1 (en) | 2019-04-18 | 2019-04-18 | Composition for improving, preventing or treating bone diseases comprising Lactobacillus curvatus WiKim38 or culture medium thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| KR1020190045533A KR102144838B1 (en) | 2019-04-18 | 2019-04-18 | Composition for improving, preventing or treating bone diseases comprising Lactobacillus curvatus WiKim38 or culture medium thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| KR102144838B1 true KR102144838B1 (en) | 2020-08-14 |
Family
ID=72050217
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| KR1020190045533A KR102144838B1 (en) | 2019-04-18 | 2019-04-18 | Composition for improving, preventing or treating bone diseases comprising Lactobacillus curvatus WiKim38 or culture medium thereof |
Country Status (1)
| Country | Link |
|---|---|
| KR (1) | KR102144838B1 (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102488905B1 (en) * | 2021-12-17 | 2023-01-18 | 주식회사 메디오젠 | Novel Lactobacillus curvatus strain, and uses thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20130132172A (en) * | 2012-05-25 | 2013-12-04 | 한국 한의학 연구원 | Pharmaceutical composition and functional food for prevention or treatment of bone disease comprising the lactic acid bacteria ferment of hwangryunhaedoktang |
| KR20140140387A (en) * | 2013-05-29 | 2014-12-09 | 바이오제닉스코리아 주식회사 | Nano-Sized Lactic Acid Bacteria from Kimchi |
| KR101680014B1 (en) * | 2016-07-04 | 2016-11-29 | 한국식품연구원 | Lactic acid bacterium isolated from Kimchi for treating inflammatory bowel disease and uses thereof |
| KR101838281B1 (en) * | 2016-12-12 | 2018-03-14 | 한국식품연구원 | Lactobacillus curvatus WIKIM53 having anti-allergic disease activity and composition for comprising the same |
-
2019
- 2019-04-18 KR KR1020190045533A patent/KR102144838B1/en active IP Right Grant
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20130132172A (en) * | 2012-05-25 | 2013-12-04 | 한국 한의학 연구원 | Pharmaceutical composition and functional food for prevention or treatment of bone disease comprising the lactic acid bacteria ferment of hwangryunhaedoktang |
| KR20140140387A (en) * | 2013-05-29 | 2014-12-09 | 바이오제닉스코리아 주식회사 | Nano-Sized Lactic Acid Bacteria from Kimchi |
| KR101680014B1 (en) * | 2016-07-04 | 2016-11-29 | 한국식품연구원 | Lactic acid bacterium isolated from Kimchi for treating inflammatory bowel disease and uses thereof |
| KR101838281B1 (en) * | 2016-12-12 | 2018-03-14 | 한국식품연구원 | Lactobacillus curvatus WIKIM53 having anti-allergic disease activity and composition for comprising the same |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR102488905B1 (en) * | 2021-12-17 | 2023-01-18 | 주식회사 메디오젠 | Novel Lactobacillus curvatus strain, and uses thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| KR102242668B1 (en) | Composition for improving, preventing or treating bone diseases comprising Lactobacillus sakei CVL-001 or culture medium thereof | |
| KR101494279B1 (en) | Lactobacillus plantarum KY1032 having inhibitory activity against adipocyte-specific gene expression and adipocyte differentiation, and product containing thereof as an effective factor | |
| US11273189B2 (en) | Lactobacillus plantarum TCI378 and its uses in losing fat and improving gastrointestinal functions | |
| KR101680014B1 (en) | Lactic acid bacterium isolated from Kimchi for treating inflammatory bowel disease and uses thereof | |
| KR102084973B1 (en) | Composition for preventing or treating colitis comprising enterococcus faecalis | |
| US20140050805A1 (en) | Composition, glucose metabolism-improving agent, and method for improving glucose metabolism | |
| KR102526564B1 (en) | Composition for preventing or treating inflammatory disease comprising Lactibacillus sakei CVL-001 strain | |
| KR102486028B1 (en) | Composition for preventing, treating or improving bone diseases containing extracellular vesicles derived from Lactobacillus sakei CVL001 strain culture medium | |
| KR20210067413A (en) | A mixture of Lactobacillus plantarum LRCC5195 and isomaltooligosaccharide having ameliorating or improving atopic dermatitis | |
| KR101402926B1 (en) | Pharmaceutical composition and functional food for prevention or treatment of bone disease comprising the lactic acid bacteria ferment of hwangryunhaedoktang | |
| KR102144838B1 (en) | Composition for improving, preventing or treating bone diseases comprising Lactobacillus curvatus WiKim38 or culture medium thereof | |
| EP3932416A2 (en) | Composition for improving, preventing, or treating bone diseases or metabolic diseases, including novel lactobacillus sakei cvl-001 strain and culture medium thereof | |
| KR102190239B1 (en) | Novel Lactobacillus plantarum Lb41 strain and compositions for the prevention and treatment of obesity containing the same | |
| EP4074324A1 (en) | Composition containing enteroccocus faecalis as active ingredient for preventing or treating obesity or metabolic syndromes induced thereby | |
| KR101504912B1 (en) | The products containing probiotic lactobacillus acidophilus HY7037 having activity preventing from insulin resistance, which caused type 2 diabetes mellitus | |
| JP6976030B2 (en) | Composition for prevention or improvement of non-alcoholic fatty liver disease | |
| RU2782361C1 (en) | Composition for alleviating, preventing, or treating bone diseases or metabolic diseases, containing a new strain of lactobacillus sakei cvl-001 and a culture medium thereof | |
| JP2009114112A (en) | Anti-osteoporosis agent | |
| KR101535077B1 (en) | The products containing probiotic Bifidobacterium lactis HY8101 having activity preventing from insulin resistance, which caused type 2 diabetes mellitus | |
| KR102470357B1 (en) | Composition for enhancing immunity comprising of lactococcus lactis Q1 | |
| JP6173850B2 (en) | Muscle bulking agent, muscle bulking agent when used together with exercise, and food and drink for muscle weight gain | |
| RU2785355C2 (en) | New lactic acid bacteria and application thereof | |
| JP2019011308A (en) | Oral composition | |
| KR20240086744A (en) | Pharmaceutical composition for alleviating or treating premenstrual syndrome containing enterococcus faecium lcm001 | |
| KR20220026201A (en) | Composition for preventing or treating of climacterium comprising Lactobacillus fermentum |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| AMND | Amendment | ||
| E601 | Decision to refuse application | ||
| X091 | Application refused [patent] | ||
| AMND | Amendment | ||
| X701 | Decision to grant (after re-examination) | ||
| GRNT | Written decision to grant |