CN110771769A - 一种辅助治疗二型糖尿病的壳寡糖复合固体饮料 - Google Patents
一种辅助治疗二型糖尿病的壳寡糖复合固体饮料 Download PDFInfo
- Publication number
- CN110771769A CN110771769A CN201911303224.9A CN201911303224A CN110771769A CN 110771769 A CN110771769 A CN 110771769A CN 201911303224 A CN201911303224 A CN 201911303224A CN 110771769 A CN110771769 A CN 110771769A
- Authority
- CN
- China
- Prior art keywords
- group
- t2dm
- mice
- chitosan oligosaccharide
- diabetes
- Prior art date
- Legal status (The legal status is an assumption and is not a legal conclusion. Google has not performed a legal analysis and makes no representation as to the accuracy of the status listed.)
- Pending
Links
- 208000001072 type 2 diabetes mellitus Diseases 0.000 title claims abstract description 22
- RQFQJYYMBWVMQG-IXDPLRRUSA-N chitotriose Chemical compound O[C@@H]1[C@@H](N)[C@H](O)O[C@H](CO)[C@H]1O[C@H]1[C@H](N)[C@@H](O)[C@H](O[C@H]2[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O2)N)[C@@H](CO)O1 RQFQJYYMBWVMQG-IXDPLRRUSA-N 0.000 title claims abstract description 18
- 235000013361 beverage Nutrition 0.000 title claims abstract description 15
- 239000007787 solid Substances 0.000 title claims abstract description 15
- 238000009098 adjuvant therapy Methods 0.000 title claims abstract description 10
- 239000002131 composite material Substances 0.000 title claims abstract description 9
- 229940054810 white kidney bean extract Drugs 0.000 claims abstract description 10
- UQZIYBXSHAGNOE-USOSMYMVSA-N Stachyose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO[C@@H]2[C@@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O2)O1 UQZIYBXSHAGNOE-USOSMYMVSA-N 0.000 claims abstract description 9
- 239000003094 microcapsule Substances 0.000 claims abstract description 9
- UQZIYBXSHAGNOE-XNSRJBNMSA-N stachyose 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[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO[C@@H]3[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O3)O)O2)O)O1 UQZIYBXSHAGNOE-XNSRJBNMSA-N 0.000 claims abstract description 9
- FYGDTMLNYKFZSV-URKRLVJHSA-N (2s,3r,4s,5s,6r)-2-[(2r,4r,5r,6s)-4,5-dihydroxy-2-(hydroxymethyl)-6-[(2r,4r,5r,6s)-4,5,6-trihydroxy-2-(hydroxymethyl)oxan-3-yl]oxyoxan-3-yl]oxy-6-(hydroxymethyl)oxane-3,4,5-triol Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1[C@@H](CO)O[C@@H](OC2[C@H](O[C@H](O)[C@H](O)[C@H]2O)CO)[C@H](O)[C@H]1O FYGDTMLNYKFZSV-URKRLVJHSA-N 0.000 claims abstract description 8
- 229920002498 Beta-glucan Polymers 0.000 claims abstract description 8
- 235000021388 linseed oil Nutrition 0.000 claims abstract description 8
- 239000000944 linseed oil Substances 0.000 claims abstract description 8
- 239000000843 powder Substances 0.000 claims abstract description 8
- 229920001661 Chitosan Polymers 0.000 claims description 33
- -1 chitosan oligosaccharide compound Chemical class 0.000 claims description 8
- 241000699670 Mus sp. Species 0.000 description 59
- 239000008280 blood Substances 0.000 description 20
- 210000004369 blood Anatomy 0.000 description 20
- 241000699666 Mus <mouse, genus> Species 0.000 description 18
- 230000000694 effects Effects 0.000 description 18
- 230000000968 intestinal effect Effects 0.000 description 15
- 241001474033 Acar Species 0.000 description 13
- 210000001035 gastrointestinal tract Anatomy 0.000 description 13
- 238000012163 sequencing technique Methods 0.000 description 13
- 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 12
- 206010012601 diabetes mellitus Diseases 0.000 description 12
- 210000004185 liver Anatomy 0.000 description 11
- 210000000056 organ Anatomy 0.000 description 11
- 241000894006 Bacteria Species 0.000 description 10
- 241001112693 Lachnospiraceae Species 0.000 description 10
- 210000005228 liver tissue Anatomy 0.000 description 10
- 238000004458 analytical method Methods 0.000 description 9
- 238000002474 experimental method Methods 0.000 description 9
- 108020004999 messenger RNA Proteins 0.000 description 9
- 241000192125 Firmicutes Species 0.000 description 8
- 239000000203 mixture Substances 0.000 description 8
- 108090000623 proteins and genes Proteins 0.000 description 8
- 210000002966 serum Anatomy 0.000 description 8
- 102100036475 Alanine aminotransferase 1 Human genes 0.000 description 7
- 108010082126 Alanine transaminase Proteins 0.000 description 7
- 241000606125 Bacteroides Species 0.000 description 7
- 241000605059 Bacteroidetes Species 0.000 description 7
- 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 7
- 108010023302 HDL Cholesterol Proteins 0.000 description 7
- 108090000472 Phosphoenolpyruvate carboxykinase (ATP) Proteins 0.000 description 7
- 102100034792 Phosphoenolpyruvate carboxykinase [GTP], mitochondrial Human genes 0.000 description 7
- 239000008103 glucose Substances 0.000 description 7
- 210000005084 renal tissue Anatomy 0.000 description 7
- 238000011160 research Methods 0.000 description 7
- 102100029077 3-hydroxy-3-methylglutaryl-coenzyme A reductase Human genes 0.000 description 6
- 108010003415 Aspartate Aminotransferases Proteins 0.000 description 6
- 102000004625 Aspartate Aminotransferases Human genes 0.000 description 6
- 101000988577 Homo sapiens 3-hydroxy-3-methylglutaryl-coenzyme A reductase Proteins 0.000 description 6
- 108010028554 LDL Cholesterol Proteins 0.000 description 6
- 102000004169 proteins and genes Human genes 0.000 description 6
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 6
- 101000957672 Homo sapiens Cytochrome P450 7A1 Proteins 0.000 description 5
- 101001051093 Homo sapiens Low-density lipoprotein receptor Proteins 0.000 description 5
- 108010074438 Sterol Regulatory Element Binding Protein 2 Proteins 0.000 description 5
- 230000003247 decreasing effect Effects 0.000 description 5
- 210000002216 heart Anatomy 0.000 description 5
- 230000002440 hepatic effect Effects 0.000 description 5
- 235000018102 proteins Nutrition 0.000 description 5
- 241000894007 species Species 0.000 description 5
- 238000012353 t test Methods 0.000 description 5
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 4
- 241000692822 Bacteroidales Species 0.000 description 4
- 102100038637 Cytochrome P450 7A1 Human genes 0.000 description 4
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 4
- 102100036264 Glucose-6-phosphatase catalytic subunit 1 Human genes 0.000 description 4
- 101710099339 Glucose-6-phosphatase catalytic subunit 1 Proteins 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- 102100024640 Low-density lipoprotein receptor Human genes 0.000 description 4
- 241000605861 Prevotella Species 0.000 description 4
- 230000009286 beneficial effect Effects 0.000 description 4
- 230000033228 biological regulation Effects 0.000 description 4
- 230000037396 body weight Effects 0.000 description 4
- 235000012000 cholesterol Nutrition 0.000 description 4
- 231100000673 dose–response relationship Toxicity 0.000 description 4
- 230000014509 gene expression Effects 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 230000004060 metabolic process Effects 0.000 description 4
- 238000000034 method Methods 0.000 description 4
- 244000005700 microbiome Species 0.000 description 4
- 230000001105 regulatory effect Effects 0.000 description 4
- 238000010186 staining Methods 0.000 description 4
- XUFXOAAUWZOOIT-SXARVLRPSA-N (2R,3R,4R,5S,6R)-5-[[(2R,3R,4R,5S,6R)-5-[[(2R,3R,4S,5S,6R)-3,4-dihydroxy-6-methyl-5-[[(1S,4R,5S,6S)-4,5,6-trihydroxy-3-(hydroxymethyl)-1-cyclohex-2-enyl]amino]-2-oxanyl]oxy]-3,4-dihydroxy-6-(hydroxymethyl)-2-oxanyl]oxy]-6-(hydroxymethyl)oxane-2,3,4-triol Chemical compound O([C@H]1O[C@H](CO)[C@H]([C@@H]([C@H]1O)O)O[C@H]1O[C@@H]([C@H]([C@H](O)[C@H]1O)N[C@@H]1[C@@H]([C@@H](O)[C@H](O)C(CO)=C1)O)C)[C@@H]1[C@@H](CO)O[C@@H](O)[C@H](O)[C@H]1O XUFXOAAUWZOOIT-SXARVLRPSA-N 0.000 description 3
- HVCOBJNICQPDBP-UHFFFAOYSA-N 3-[3-[3,5-dihydroxy-6-methyl-4-(3,4,5-trihydroxy-6-methyloxan-2-yl)oxyoxan-2-yl]oxydecanoyloxy]decanoic acid;hydrate Chemical compound O.OC1C(OC(CC(=O)OC(CCCCCCC)CC(O)=O)CCCCCCC)OC(C)C(O)C1OC1C(O)C(O)C(O)C(C)O1 HVCOBJNICQPDBP-UHFFFAOYSA-N 0.000 description 3
- 102000004190 Enzymes Human genes 0.000 description 3
- 108090000790 Enzymes Proteins 0.000 description 3
- 229930186217 Glycolipid Natural products 0.000 description 3
- 241001465754 Metazoa Species 0.000 description 3
- 241000692844 Prevotellaceae Species 0.000 description 3
- 229960002632 acarbose Drugs 0.000 description 3
- XUFXOAAUWZOOIT-UHFFFAOYSA-N acarviostatin I01 Natural products OC1C(O)C(NC2C(C(O)C(O)C(CO)=C2)O)C(C)OC1OC(C(C1O)O)C(CO)OC1OC1C(CO)OC(O)C(O)C1O XUFXOAAUWZOOIT-UHFFFAOYSA-N 0.000 description 3
- 244000052616 bacterial pathogen Species 0.000 description 3
- 210000001072 colon Anatomy 0.000 description 3
- 230000000875 corresponding effect Effects 0.000 description 3
- 239000003814 drug Substances 0.000 description 3
- 229940088598 enzyme Drugs 0.000 description 3
- 230000004054 inflammatory process Effects 0.000 description 3
- 230000008944 intestinal immunity Effects 0.000 description 3
- 208000030159 metabolic disease Diseases 0.000 description 3
- 239000000047 product Substances 0.000 description 3
- 239000000243 solution Substances 0.000 description 3
- 231100000240 steatosis hepatitis Toxicity 0.000 description 3
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- LKAPTZKZHMOIRE-KVTDHHQDSA-N (2s,3s,4s,5r)-3,4-dihydroxy-5-(hydroxymethyl)oxolane-2-carbaldehyde Chemical compound OC[C@H]1O[C@H](C=O)[C@@H](O)[C@@H]1O LKAPTZKZHMOIRE-KVTDHHQDSA-N 0.000 description 2
- 201000001320 Atherosclerosis Diseases 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-M Butyrate Chemical group CCCC([O-])=O FERIUCNNQQJTOY-UHFFFAOYSA-M 0.000 description 2
- FERIUCNNQQJTOY-UHFFFAOYSA-N Butyric acid Natural products CCCC(O)=O FERIUCNNQQJTOY-UHFFFAOYSA-N 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- 241000588921 Enterobacteriaceae Species 0.000 description 2
- WZUVPPKBWHMQCE-UHFFFAOYSA-N Haematoxylin Chemical compound C12=CC(O)=C(O)C=C2CC2(O)C1C1=CC=C(O)C(O)=C1OC2 WZUVPPKBWHMQCE-UHFFFAOYSA-N 0.000 description 2
- 206010019708 Hepatic steatosis Diseases 0.000 description 2
- 238000012449 Kunming mouse Methods 0.000 description 2
- 241001468155 Lactobacillaceae Species 0.000 description 2
- 206010067125 Liver injury Diseases 0.000 description 2
- 102100026841 Sterol regulatory element-binding protein 2 Human genes 0.000 description 2
- ZSJLQEPLLKMAKR-UHFFFAOYSA-N Streptozotocin Natural products O=NN(C)C(=O)NC1C(O)OC(CO)C(O)C1O ZSJLQEPLLKMAKR-UHFFFAOYSA-N 0.000 description 2
- 230000002159 abnormal effect Effects 0.000 description 2
- 238000010521 absorption reaction Methods 0.000 description 2
- 239000003472 antidiabetic agent Substances 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- LKAPTZKZHMOIRE-UHFFFAOYSA-N chitose Natural products OCC1OC(C=O)C(O)C1O LKAPTZKZHMOIRE-UHFFFAOYSA-N 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 238000001816 cooling Methods 0.000 description 2
- 238000007405 data analysis Methods 0.000 description 2
- 230000000593 degrading effect Effects 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 229940079593 drug Drugs 0.000 description 2
- 230000004110 gluconeogenesis Effects 0.000 description 2
- 230000004153 glucose metabolism Effects 0.000 description 2
- 208000018914 glucose metabolism disease Diseases 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 230000002218 hypoglycaemic effect Effects 0.000 description 2
- 239000007924 injection Substances 0.000 description 2
- 238000002347 injection Methods 0.000 description 2
- 210000003734 kidney Anatomy 0.000 description 2
- 230000003902 lesion Effects 0.000 description 2
- 230000006372 lipid accumulation Effects 0.000 description 2
- 230000037356 lipid metabolism Effects 0.000 description 2
- 150000002632 lipids Chemical class 0.000 description 2
- 239000007788 liquid Substances 0.000 description 2
- 210000005229 liver cell Anatomy 0.000 description 2
- 238000005259 measurement Methods 0.000 description 2
- 210000004379 membrane Anatomy 0.000 description 2
- 239000012528 membrane Substances 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 229910052757 nitrogen Inorganic materials 0.000 description 2
- 210000000633 nuclear envelope Anatomy 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 238000003908 quality control method Methods 0.000 description 2
- 230000002441 reversible effect Effects 0.000 description 2
- 230000000087 stabilizing effect Effects 0.000 description 2
- 238000007619 statistical method Methods 0.000 description 2
- 230000007863 steatosis Effects 0.000 description 2
- 229960001052 streptozocin Drugs 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 210000003384 transverse colon Anatomy 0.000 description 2
- 210000003462 vein Anatomy 0.000 description 2
- 230000009278 visceral effect Effects 0.000 description 2
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 2
- 208000016261 weight loss Diseases 0.000 description 2
- 230000004580 weight loss Effects 0.000 description 2
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- 241001468182 Acidobacterium Species 0.000 description 1
- 102000007469 Actins Human genes 0.000 description 1
- 108010085238 Actins Proteins 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 101710171801 Alpha-amylase inhibitor Proteins 0.000 description 1
- 241000252983 Caecum Species 0.000 description 1
- 241000372691 Candidatus Saccharibacteria Species 0.000 description 1
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 241000192700 Cyanobacteria Species 0.000 description 1
- 108020004414 DNA Proteins 0.000 description 1
- 241000521195 Deferribacter Species 0.000 description 1
- 208000005156 Dehydration Diseases 0.000 description 1
- 241000605716 Desulfovibrio Species 0.000 description 1
- 241001467894 Desulfovibrionaceae Species 0.000 description 1
- 229920002307 Dextran Polymers 0.000 description 1
- 208000036649 Dysbacteriosis Diseases 0.000 description 1
- 208000027244 Dysbiosis Diseases 0.000 description 1
- 208000004930 Fatty Liver Diseases 0.000 description 1
- 206010016654 Fibrosis Diseases 0.000 description 1
- 241000605909 Fusobacterium Species 0.000 description 1
- 102000003886 Glycoproteins Human genes 0.000 description 1
- 108090000288 Glycoproteins Proteins 0.000 description 1
- 206010019668 Hepatic fibrosis Diseases 0.000 description 1
- 208000031226 Hyperlipidaemia Diseases 0.000 description 1
- 208000017170 Lipid metabolism disease Diseases 0.000 description 1
- 241001467578 Microbacterium Species 0.000 description 1
- 206010028980 Neoplasm Diseases 0.000 description 1
- 238000012408 PCR amplification Methods 0.000 description 1
- 239000012807 PCR reagent Substances 0.000 description 1
- 241000425347 Phyla <beetle> Species 0.000 description 1
- 241000233614 Phytophthora Species 0.000 description 1
- 241000192142 Proteobacteria Species 0.000 description 1
- 241001135259 Rikenella Species 0.000 description 1
- 241000692845 Rikenellaceae Species 0.000 description 1
- 241000192031 Ruminococcus Species 0.000 description 1
- 206010040744 Sinus headache Diseases 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 108091023040 Transcription factor Proteins 0.000 description 1
- 102000040945 Transcription factor Human genes 0.000 description 1
- 241000700605 Viruses Species 0.000 description 1
- 239000013543 active substance Substances 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 230000003321 amplification Effects 0.000 description 1
- 239000003392 amylase inhibitor Substances 0.000 description 1
- 229940127003 anti-diabetic drug Drugs 0.000 description 1
- 230000003178 anti-diabetic effect Effects 0.000 description 1
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 210000001815 ascending colon Anatomy 0.000 description 1
- 230000001580 bacterial effect Effects 0.000 description 1
- 230000003115 biocidal effect Effects 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000000740 bleeding effect Effects 0.000 description 1
- 229910052799 carbon Inorganic materials 0.000 description 1
- 210000004534 cecum Anatomy 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 229920002678 cellulose Polymers 0.000 description 1
- 239000001913 cellulose Substances 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 108010089807 chitosanase Proteins 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 239000000470 constituent Substances 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 238000013211 curve analysis Methods 0.000 description 1
- 238000005520 cutting process Methods 0.000 description 1
- 230000003013 cytotoxicity Effects 0.000 description 1
- 231100000135 cytotoxicity Toxicity 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 230000007850 degeneration Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000011161 development Methods 0.000 description 1
- 238000013118 diabetic mouse model Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 235000019621 digestibility Nutrition 0.000 description 1
- 230000008034 disappearance Effects 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000002651 drug therapy Methods 0.000 description 1
- 230000007140 dysbiosis Effects 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 208000010706 fatty liver disease Diseases 0.000 description 1
- 230000002550 fecal effect Effects 0.000 description 1
- 230000004761 fibrosis Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 239000012634 fragment Substances 0.000 description 1
- 235000013376 functional food Nutrition 0.000 description 1
- 230000007160 gastrointestinal dysfunction Effects 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 238000001502 gel electrophoresis Methods 0.000 description 1
- 229960004580 glibenclamide Drugs 0.000 description 1
- 210000005086 glomerual capillary Anatomy 0.000 description 1
- 230000001434 glomerular Effects 0.000 description 1
- ZNNLBTZKUZBEKO-UHFFFAOYSA-N glyburide Chemical compound COC1=CC=C(Cl)C=C1C(=O)NCCC1=CC=C(S(=O)(=O)NC(=O)NC2CCCCC2)C=C1 ZNNLBTZKUZBEKO-UHFFFAOYSA-N 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 231100000234 hepatic damage Toxicity 0.000 description 1
- 231100000753 hepatic injury Toxicity 0.000 description 1
- 235000009200 high fat diet Nutrition 0.000 description 1
- 238000012165 high-throughput sequencing Methods 0.000 description 1
- 201000001421 hyperglycemia Diseases 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 238000003331 infrared imaging Methods 0.000 description 1
- 210000004347 intestinal mucosa Anatomy 0.000 description 1
- 230000003870 intestinal permeability Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 230000008818 liver damage Effects 0.000 description 1
- 239000012139 lysis buffer Substances 0.000 description 1
- 238000010841 mRNA extraction Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 230000006371 metabolic abnormality Effects 0.000 description 1
- XZWYZXLIPXDOLR-UHFFFAOYSA-N metformin Chemical compound CN(C)C(=N)NC(N)=N XZWYZXLIPXDOLR-UHFFFAOYSA-N 0.000 description 1
- 229960003105 metformin Drugs 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 238000000465 moulding Methods 0.000 description 1
- 230000004682 mucosal barrier function Effects 0.000 description 1
- 201000001119 neuropathy Diseases 0.000 description 1
- 230000007823 neuropathy Effects 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 238000003199 nucleic acid amplification method Methods 0.000 description 1
- 235000015097 nutrients Nutrition 0.000 description 1
- 229920001542 oligosaccharide Polymers 0.000 description 1
- 238000005457 optimization Methods 0.000 description 1
- 229940127017 oral antidiabetic Drugs 0.000 description 1
- 230000008816 organ damage Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 208000033808 peripheral neuropathy Diseases 0.000 description 1
- 239000002504 physiological saline solution Substances 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 235000013406 prebiotics Nutrition 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000000513 principal component analysis Methods 0.000 description 1
- 230000035755 proliferation Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 238000011002 quantification Methods 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011084 recovery Methods 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 239000003053 toxin Substances 0.000 description 1
- 231100000765 toxin Toxicity 0.000 description 1
- 108700012359 toxins Proteins 0.000 description 1
- 238000009827 uniform distribution Methods 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Images
Classifications
-
- 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
- A23L2/00—Non-alcoholic beverages; Dry compositions or concentrates therefor; Their preparation
- A23L2/385—Concentrates of non-alcoholic beverages
- A23L2/39—Dry compositions
-
- 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
- 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
- A23L33/115—Fatty acids or derivatives thereof; Fats or oils
-
- 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
- A23L33/20—Reducing nutritive value; Dietetic products with reduced nutritive value
- A23L33/21—Addition of substantially indigestible substances, e.g. dietary fibres
-
- 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
Landscapes
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Health & Medical Sciences (AREA)
- Nutrition Science (AREA)
- Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Mycology (AREA)
- Botany (AREA)
- Oil, Petroleum & Natural Gas (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
Abstract
本发明涉及一种辅助治疗二型糖尿病的壳寡糖复合固体饮料，按重量百分数计，所述壳聚糖复合固体饮料包括以下组分：壳寡糖5～30％；水苏糖2～18％；燕麦β‑葡聚糖5～20％；白芸豆提取物16～38％；亚麻籽油微囊粉10～20％。该壳寡糖复合固体饮料具有辅助降低血脂及稳定血糖的作用，同时，该固体饮料可以改善肠道菌群、提高肠道免疫力以辅助二型糖尿病。
Description
技术领域
本发明涉及保健品技术领域，尤其是涉及一种辅助治疗二型糖尿病的壳寡糖复合固体饮料。
背景技术
二型糖尿病(Type 2diabetes，T2DM)是一种以高血糖为特征 (往往还合并高血脂症状)的代谢紊乱症之一，常伴随如神经性病变、肾病、心脑血管疾病等多种慢性并发症发生。可以导致多器官、多系统的损伤致残，致死率高。它是公共卫生的主要威胁，据国际糖尿病联合会估计，2015年全球有4.15亿糖尿病患者，到2035年这一数字预计将增加到5.92亿，而我国糖尿病患者远超于其他国家，并且人数还在持续增加。因此，我国在糖尿病的防治方面面临着巨大的挑战。
传统的抗糖尿病策略主要是药物治疗结合饮食及运动调节。目前，二甲双胍、格列本脲、阿卡波糖等为临床上常见的口服抗糖尿病药物。然而，这些可用的抗糖尿病药物尚存在一些器官损伤或胃肠道功能障碍等不良反应。因此，寻找天然的，特别是食品源的降血糖活性物质，开新的降血糖功能食品依然是大健康研究领域的重要热点之一。
发明内容
为了克服现有技术存在的上述技术问题，本发明提供一种辅助治疗二型糖尿病的壳寡糖复合固体饮料，该固体饮料以壳寡糖、水苏糖、燕麦β-葡聚糖、白芸豆提取物等为主要原料，具有辅助降低血脂及稳定血糖的作用；同时，该固体饮料可以改善肠道菌群、提高肠道免疫力以辅助二型糖尿病。
一种辅助治疗二型糖尿病的壳寡糖复合固体饮料，所述壳聚糖复合固体饮料包括壳寡糖、水苏糖、燕麦β-葡聚糖、白芸豆提取物和亚麻籽油微囊粉。
壳寡糖是由壳聚糖降解而得到，是碱性氨基低聚糖，是动物性纤维素制备的益生元，具有很多生物活性，比如低细胞毒性、水溶性好、易于被肠道吸收、抗氧化、抗炎、辅助治疗二型糖尿病、抗菌、降胆固醇、调节免疫和抗肿瘤等。
燕麦β-葡聚糖和水苏糖都是功能性的糖类，具有很多生理功能，比如促进肠道内有益菌的增殖继而提高其所占比例、抑制肠道内有害菌的生长、阻止外源性病原菌在肠道的定植、增加机体肠道免疫力，通过改善肠道微生态而发挥其功能性活性。白芸豆提取物中的α-淀粉酶抑制剂属于天然糖蛋白，既安全绿色又没有任何毒性，可以通过降低淀粉类的消化率，进入肠道后为肠道微生物提供优质碳源，来辅助降低2型糖尿病病人的血糖。
进一步的，按重量百分数计，所述壳聚糖复合固体饮料中各组分的含量如下：壳寡糖5～30％；水苏糖2～18％；燕麦β-葡聚糖5～20％；白芸豆提取物16～38％；亚麻籽油微囊粉10～20％。
进一步的，其特征在于，按重量百分数计，壳寡糖28％；水苏糖 18％；燕麦β-葡聚糖18％；白芸豆提取物16％；亚麻籽油微囊粉20％。
相对现有技术，本发明的有益效果在于：
(1)壳寡糖复合固体饮料具有辅助降低T2DM小鼠血脂及稳定血糖的作用。
(2)壳聚糖复合固体饮料可以影响肠道菌群的组成和多样性，并且促进有益菌的生长与定植，降低条件致病菌的相对丰度，从而促进肠道健康，并可辅助治疗二型糖尿病等疾病。
附图说明
为了更清楚地说明本发明实施例或现有技术中的技术方案，下面将对实施例或现有技术描述中所需要使用的附图作简单地介绍，显而易见地，下面描述中的附图仅仅是本发明的一些实施例，对于本领域普通技术人员来讲，在不付出创造性劳动的前提下，还可以根据这些附图获得其他的附图。
图1不同剂量壳寡糖复合固体饮料对小鼠血糖影响变化；
图2小鼠肝脏和肾组织H&E染色；
图3壳糖平调节葡萄糖代谢中关键基因mRNA水平和蛋白水平。
图4(A)稀释曲线(B)Shannon-Wiener曲线分析。
图5七组样本在门水平上的相对丰度。
图6七组样本在科水平上的相对丰度。
图7各实验组的LEFse分析比较。
图8七组样本OTU水平上的维恩图。
图9小鼠肠道样本PCoA分析
其中：
图2中(A)肝脏组织H&E染色(20×)；(B)肾脏组织H&E染色(20×)；
图3中(A)PEPCK mRNA水平，(B)HMGCR mRNA水平， (C)G6pase mRNA水平，(D)SREBP-2mRNA水平，(E)CYP7A1 mRNA水平，(F)LDLR mRNA水平，(G)PEPCK、HMGCR蛋白水平。注：经t检验，#表示与NFD组比较差异显著(p<0.05)， ##表示与NFD组比较差异极显著(p<0.01)；*表示与T2DM组比较差异显著(p<0.05)，**表示与T2DM组比较差异极显著(p<0.01)，n＝8。
具体实施方式
下面结合具体实施例对本发明做进一步的分析，显然，所描述的仅仅是本发明一部分实施例，而不是全部的实施例。基于本发明中的实施例，本领域普通技术人员在没有作出创造性劳动的前提下所获得的所有其他实施例，都属于本发明保护的范围。
为了方便，本发明下述实施例和实验例中将“壳寡糖复合固体饮料”简称为“壳糖平”。
实施例
| 壳糖平的组分 | 实施例1 | 实施例2 | 实施例3 | 实施例4 |
| 壳寡糖 | 5％ | 30％ | 28％ | 26％ |
| 水苏糖 | 17％ | 2％ | 18％ | 15％ |
| 燕麦β-葡聚糖 | 20％ | 20％ | 18％ | 5％ |
| 白芸豆提取物 | 38％ | 38％ | 16％ | 36％ |
| 亚麻籽油微囊粉 | 20％ | 10％ | 20％ | 18％ |
实验例
为了进一步说明效果，以实施例3为例进行实验，具体实验情况如下：
1、实验材料和仪器
实验用的壳糖平以实施例3为准；4周龄(体重18～22g)健康雄性清洁级昆明小鼠【生产许可证：SCXK(军)2007-004，合格证号： 00549571】及高脂饲料和基础饲料购自中国人民解放军军事医学科学院实验动物中心；阿卡波糖片(Acar)杭州中美华东制药有限公司；总胆固醇(TC)、甘油三酯(TG)、高密度脂蛋白胆固醇(HDL-C)、低密度脂蛋白胆固醇(LDL-C)、谷草转氨酶(AST)、谷丙转转氨酶(ALT)试剂盒南京建成生物工程研究所；AxyPrepDNA Gel Ext raction Kit试剂盒购自AXYGEN公司；KOD Polymerase购自Illumi na公司。
2、实验方法
2.1实验分组和二型糖尿病小鼠模型的建立
健康雄性SPF昆明小鼠70只(4周龄，20±2g)。按照体重随机分组分为7组，每组10只，分组和饲喂剂量为：正常对照组(NF D+PBS)、模型组(T2DM+PBS)、阳性对照阿卡波糖(50mg/kg) 组(T2DM+Acar)、壳寡糖(80mg/kg)组(T2DM+COS)、壳糖平低剂量(40mg/kg)组(T2DM+LKTP)、壳糖平中剂量(80mg/ kg)组(T2DM+MKTP)、壳糖平高剂量(120mg/kg)组(T2DM+ HKTP)。实验小鼠以基础饲料适应性喂养一周后，T2DM、T2DM+ Acar、T2DM+COS、T2DM+LKTP、T2DM+MKTP和T2DM+HKTP 组小鼠禁食不禁水12h，采用65mg/kg链脲佐菌素(STZ)连续3d进行腹腔注射，造模期间饲喂高脂饲料。注射药物5d后，小鼠尾静脉取血，测定小鼠的血糖；据报告：空腹血糖水平≥7.0mmol/L，餐后2h血糖水平≥11.1mmol/L为造模成功，从第4周开始分别灌胃4周。
2.2壳糖平对二型糖尿病小鼠糖脂代谢紊乱的调节作用的实验方法
2.2.1小鼠血液样品及器官的处理与保存
实验结束后，小鼠禁食12h，采取摘眼球法活体取血，取血完毕后采用断颈法处死小鼠。将血液放室温平衡10min，3000rpm离心10min，取上清，-20℃保存，对小鼠进行解剖，摘取小鼠的肝脏和心脏，用无菌生理盐水冲洗后吸干水分后快速称重，计算肝脏和心脏指数。参考现有文献的方法，其计算公式为：脏器指数(％)＝脏器重量/体重×100；同时将摘取的小鼠肝脏、心脏、升结肠、横结肠、肾脏组织分装于对应的离心管中，其中脏器组织的一部分迅速至于液氮罐中冷却，保存于超低温(-80℃)冰箱中。取脏器组织块少量置于含福尔马林溶液的离心管中，常温保存备用。
2.2.2小鼠血糖和血清各指标的检测
第3周开始直到结束期间每周测定一次空腹血糖。采用尾静脉取血，测定其空腹血糖值。将2.2.1保存的血清参照试剂盒说明书进行 TC、TG、HDL-C和LDL-C、AST、ALT含量的测定。根据公式计算动脉动脉粥样硬化指数：AI＝(TC-HDL-C)/HDL-C。
2.2.3肝脏组织和肾脏组织HE染色
将2.2.2中固定在福尔马林溶液中的肝肾组织块取出进行脱水处理，用石蜡进行包埋并切片，用苏木精等进行染色，在正置显微镜观察并获取照片。
2.2.4肝脏mRNA提取和检测
将2.2.2保存的小鼠肝脏称取30mg，提取总RNA并逆转录成 cDNA，使用表1设计的引物以SYBR Green PCR试剂检测各目的基因的mRNA水平。以β-肌动蛋白为参照并通过
方法计算结果
表1实时定量检测所设计引物序列
2.2.5蛋白质印迹分析
用蛋白质裂解缓冲液在冰上裂解肝脏组织30min。使用SDS-P AGE凝胶电泳分离蛋白质，然后通过电印迹法转移到NC膜，将膜与相应的一抗4℃过夜孵育，再用相应的二抗抗体作用膜。通过红外成像系统使目的蛋白成像可视。
2.2.6数据分析
数据采用软件SPSS 17.0进行统计学分析。实验结果表示为平均值±标准偏差(M±SD)。P<0.05时，表示与比较组存在显著差异， P<0.01表示与比较组差异极显著。
2.3壳糖平对二型糖尿病小鼠肠道菌群结构的影响的实验方法
2.3.1小鼠的处理与组织保存
实验结束后，小鼠禁食12h，采用断颈法处死小鼠。进行解剖摘取小鼠横结肠部分装于离心管中，迅速置于液氮罐中冷却，保存于超低温冰箱(-80℃)中备用。
2.3.2小鼠结肠内容物菌群DNA提取和16S测序
将保存在超低温冰箱中的结肠内容物取出解冻。利用DNA提取试剂盒提取盲肠内容物微生物总DNA。用带有barcode的特异引物扩增16S rDNA的V3+V4区。引物序列为:：341F：CCTACGGGNG GCWGCAG；806R：GGACTACHVGGGTATCTAAT。然后进行PCR 扩增产物，运用切胶回收试剂盒回收目的片段，并进行定量。将纯化的扩增产物进行等量混合，连接测序接头，构建测序文库，运用His eq2500 PE250上机测序。
2.3.3数据分析
用高通量Illumina Hiseq 2500测序得到原始数据，将不可信和低质量的数据过滤掉，继而得到有效可信的序列。随后将有效序列进行拼接、OTU聚类和物种分类学分析。在OTU基础上聚类分析结果，在各个分类水平上进行群落结构的统计分析。另一方面，基于PCAO 距离，采用主坐标分析对样本进行加权和聚类，获得各样本的菌群差异信息，上述统计分析采SPSS17.0软件进行，P<0.05为有统计学意义差异。
3、实验结果
3.1壳糖平对二型糖尿病小鼠糖脂代谢紊乱的调节作用的实验结果
3.1.1壳糖平对T2DM小鼠体重的影响
由表2可以看出，与对照组相比，其余各组小鼠体重显著降低(p <0.05或p<0.01)，说明糖尿病小鼠“三多一少”中体重减少明显，符合糖尿病发病特征。与T2DM模型组比，壳糖平治疗组以剂量依赖方式显著减缓小鼠体重降低(p<0.05或p<0.01)。该结果表明壳糖平能够减缓T2DM小鼠体重减轻的症状。
表2各组小鼠实验前后体重变化(M±SD，n＝10)
注：经t检验，a表示与NFD组比较差异显著(p<0.05)，aa表示与NFD组比较差异显著(p<0.01)； b表示与T2DM组比较差异显著(p<0.05)，bb表示与T2DM组比较差异显著(p<0.01)。
3.1.2壳糖平对T2DM小鼠脏器指数的影响
由表3可以看出，与对照组(NFD)小鼠相比，其余各组小鼠肝脏指数和心脏指数指数显著增大(p<0.05或p<0.01)，脏器指数能够反应脏器的健康状况，二型糖尿病加高脂饮食会使得小鼠器脏发生脂质堆积及发生病变，继而使得脏器指数增大。与模型组(T2DM)相比，壳糖平以剂量依赖方式显著降低T2DM小鼠肝脏和心脏指数(p <0.05或p<0.01)。该结果表明壳糖平治疗能够显著逆转T2DM小鼠脏器脂质堆积和病变。
表3壳糖平对T2DM小鼠脏器指数的影响(M±SD，n＝10)
注：经t检验，a表示与NFD组比较差异显著(p<0.05)，aa表示与NFD组比较差异极显著(p<0.01)； b表示与T2DM组比较差异显著(p<0.05)，bb表示与T2DM组比较差异极显著(p<0.01)。
3.1.3壳糖平对T2DM小鼠血糖及血脂指标的影响
由图1可以看出，与对照组(NFD)小鼠相比，其余各组小鼠血糖显著升高(p<0.05或p<0.01)，说明二型糖尿病(T2DM)模型小鼠成功建立。与模型组(T2DM)相比，壳糖平治疗组以剂量依赖方式显著降低T2DM小鼠血糖(p<0.05或p<0.01)，并且随实验时间延长稳步下降。该结果表明壳糖平可有效降低T2DM小鼠血糖。
由表4可知，与对照组相比，二型糖尿病(T2DM)模型组小鼠的TG、TC、LDL-C显著上升，而HDL-C下降(p<0.05或p<0.01)，表明小鼠血清脂质水平发生改变，二型糖尿病小鼠模型造模成功。与 T2DM模型组相比，壳糖平治疗显著降低血清中TG、TC、LDL-C水平,而提高HDL-C水平((p<0.05或p<0.01)；说明壳糖平治疗具有调节T2DM小鼠脂质代谢紊乱的作用。
并且，与模型组相比，壳糖平治疗显著降低AI值(p<0.05或p <0.01)，表明壳糖平治疗能够减少动脉粥样硬化相关疾病的发生概率。
表4壳糖平对T2DM小鼠血清TC、TG、HDL-C和LDL-C的含量的影响(M±SD，n＝10)
注：经t检验，a表示与NFD组比较差异显著(p<0.05)，a表示与NFD组比较差异极显著(p<0.01)；b 表示与T2DM组比较差异显著(p<0.05)，bb表示与T2DM组比较差异极显著(p<0.01)；c表示与T2DM+Acar 组比较差异显著(p<0.05)，cc表示与T2DM+Acar组比较差异极显著(p<0.01)。
3.1.4壳糖平对T2DM小鼠血清AST、ALT的含量的影响
由表5可知，与对照组(NFD)小鼠相比，其余各组小鼠血清A ST、ALT水平指数显著增大(p<0.05或p<0.01)；T2DM小鼠糖代谢障碍，导致脂肪在肝脏中蓄积,引起脂肪肝，造成肝损伤；与模型组(T2DM)相比，壳糖平治疗组以剂量依赖方式显著降低血清AST、 ALT水平(p<0.05或p<0.01)；说明壳糖平治疗能够显著恢复T2D M小鼠肝损伤。
表5壳糖平对T2DM小鼠血清AST、ALT的含量的影响(M±SD，n＝10)
注：经t检验，a表示与NFD组比较差异显著(p<0.05)，aa表示与NFD组比较差异极显著(p<0.01)； b表示与T2DM组比较差异显著(p<0.05)，bb表示与T2DM组比较差异极显著(p<0.01)。
3.1.5壳糖平对T2DM小鼠肝和肾组织的影响
小鼠肝和肾组织H&E染色结果如图2(A)所示，NFD组小鼠肝细胞形态、大小正常、核膜核仁清楚、核圆居中、肝小叶结构正常，肝板之间分界明显，肝窦清晰可见；T2DM模型组小鼠肝组织结构出现病理变化，如肝窦受压变窄甚至消失、肝细胞大小不一，核膜核仁模糊、肝索排列无序，肝纤维化和脂肪变性严重。同样的，如图2(B) 所示，与NFD组小鼠肾切片相比较，模型组(T2DM)小鼠肾小球体积增大，细胞数增多，肾小球毛细血管扩张，出现空泡变性。而给予壳糖平各组肝脏和肾脏脂肪变性症状及纤维化得到一定程度的恢复。该结果说明壳糖平具有良好保护肝脏和肾脏、缓解糖尿病的作用。 3.1.6壳糖平对T2DM小鼠肝组织mRNA水平和蛋白水平的脂质代谢影响
如图3所示：为了探讨壳糖平对T2DM诱导的异常葡萄糖代谢的潜在机制，从mRNA和蛋白水平评估小鼠肝脏组织中PEPCK、G6 Pase、HMGCR、SREBP-2、CYP7A1、LDLR基因的变化情况。其中 PEPCK和G6Pase是肝糖异生关键酶，可以调节糖异生；HMGCR 是胆固醇合成的限速酶，而CYP7A1是一种参与胆固醇合成的的酶， LDLR和SREBP-2基因是脂类物质代谢的关键转录因子。
与正常组(NFD)相比，模型组(T2DM)小鼠PEPCK、G6Pas e、HMGCR、SREBP-2基因显著上调，CYP7A1和LDLR显著下调，说明T2DM组小鼠葡萄糖代谢紊乱同时伴随着胆固醇代谢异常，因此PEPCK、G6Pase、HMGCR、SREBP-2基因表达被异常激活，而L DLR和CYP7A1基因表达被抑制。
与T2DM组相比，壳糖平治疗可降低PEPCK、G6Pase、HMGC R、SREBP-2基因表达，而上调LDLR和CYP7A1基因表达(p<0.05 或p<0.01)。该结果表明壳糖平可以改善葡萄糖代谢紊乱和胆固醇代谢异常，维持机体内糖脂代谢平衡。
3.2壳糖平对二型糖尿病小鼠肠道菌群结构的影响的实验结果
3.2.1测序数据得质控与统计
运用Hiseq 2500二代高通量测序仪获得小鼠肠道样本16S测序原始数据后，经序列质控优化及统计后得到每个实验组的有效序列为130301～244496，其长度范围在315～478bp。为了研究各组小鼠肠道微生物菌群占比和组成多样性信息，用Uparse软件对样品进行有效序列聚类，将一致性达97％序列聚类成OTUs(Operational Taxonomic Units)，本研究中各实验组OTU范围为1367～1835，肠道菌群的物种覆盖率大于0.99，表明本研究测序覆盖率和深度可满足样品16S测序分析需求。由图4(A)可知，7组小鼠结肠样品的稀释曲线在10 0000趋向于平坦，说明本研究样品测序数据可靠测序深度满足要求；如图4(B)Shannon-Wiener曲线趋向平坦，说明测序数据物种丰富度高且分布均匀，其中NFD组样品群落多样性最高，其它依次是C OS组、MKTP组、T2DM组、LKTP组、HKTP组、Acar组，并且 (表6)NFD组ACE指数(2720.99)、chao1指数(2745.55)、S hannon香农指数(7.31)值都是最高的，说明高脂联合STZ诱导的T 2DM小鼠肠道菌群发生了显著变化，使得其肠道菌群的多样性降低，而灌服COS、LKTP和MKTP提高了Shannon香农指数，说明灌服壳寡糖及壳糖平可有效恢复T2DM小鼠肠道菌群多样性。
表6各组样品有OTUs数量和菌群多样性
3.2.2在门水平上的菌群丰度及差异
采用Hiseq高通量测序平台检测微生物组成和多样性，将检测到的微生物由门到种进行各个水平上的分类，发现建模组T2DM小鼠粪便菌群与各受试物组的粪便菌群存显著差异。如图5，7组小鼠在门(phylum)的水平上的菌群组成主要由厚壁菌门(Firmicutes)、拟杆菌门(Bacteroidetes)、变行菌门(Proteobaterria)、酸杆菌门(A ctinobateria)、蓝菌门(Cyanobacteria)、Saccharibacteria、疣微菌(V errucomicrobia)、梭杆菌门(Fusobacteria)、脱铁杆菌门(Deferrib acteres)、浮霉菌门(Planctomycetes)等10个菌门组成。可以明显看出，厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes)为主要优势菌，这与Larsen等研究报道一致；与NFD组相比(占比为38.33％)，在T2DM组中厚壁菌门(Firmicutes)显著升高(占比54.57％，p<0. 05)；而Acar组、COS组、LKTP组、MKTP组、HKTP组与T2D M模型组相比均可显著降低厚壁菌门丰度(p<0.05)，其中Acar组占比28.61％、COS组占比30.40％、LKTP组占比20.00％、MKTP 组占比20.76％、HKTP组占比23.89％；与NFD组相比(占比为56. 31％)，拟杆菌门(Bacteroidetes)丰度在T2DM组中显著降低(占比40.12％)；与T2DM模型组相比，其他给药组拟杆菌门丰度均显著升高(p<0.05)，其中MKTP组占比最高(75.88％)、其次是LK TP组占比74.27％、HKTP组占比67.64％、COS组占比65.90％、Ac ar组占比64.48％。研究表明，厚壁菌门(Firmicutes)和拟杆菌门(Bacteroidetes)协同作用于宿主能量吸收或代谢；而糖尿病患者肠道与正常人肠道对比拟杆菌门(Bacteroidetes)显著性降低，厚壁菌门(F irmicutes)显著性降低。
该结果表明：高脂联合STZ诱导的二型糖尿病小鼠厚壁菌门(F irmicutes)和拟杆菌门(Bacteroidetes)发生变化，而灌服壳糖平对二型糖尿病小鼠肠道菌群失调具有一定改善调节作用，其中壳糖平中剂量组作用效果最显著。
3.2.3在科平上的菌群组成及差异分析
如图6，7组样本在科分类水平上的主要组成菌为：拟杆菌目(B acteroidales_S24-7_group)、毛螺科菌(Lachnospiraceae)、理研菌科(Rikenellaceae)、普雷沃氏菌(Prevotellaceae)、瘤胃菌科(Ru minococcaceae)、拟杆菌科(Bacteroidaceae)、脱疏弧菌科(Desul fovibrionaceae)、肠杆菌科(Enterobacteriaceae)、乳酸杆菌科(Lactobacillaceae)、紫单胞菌科(Porphyromonadaceae)等10个菌科；其中拟杆菌目(Bacteroidales_S24-7_group)、毛螺科菌(Lachnospi raceae)为两种丰度最大的优势菌，占比超过了57％。与正常对照组 (NFD)相比，T2DM模型组拟杆菌目(Bacteroidales_S24-7_group) 和普雷沃氏菌(Prevotellaceae)显著下降(p<0.05)，而毛螺科菌(Lachnospiraceae)和脱硫弧菌科(Desulfovibrionaceae)显著升高(p <0.05)。与模型组(T2DM)相比，Acar组、COS组、LKTP组、M KTP组、HKTP组拟杆菌目(Bacteroidales_S24-7_group)、普雷沃氏菌(Prevotellaceae)显著升高(p<0.05)，而毛螺科菌(Lachnospiraceae)和脱硫弧菌科(Desulfovibrionaceae)显著降低(p<0.05)，其中HKTP组Bacteroidales_S24-7_group升高最明显，而Lachnospir aceae降低最明显。与阳性药组(Acar)相比(图7)，通过LEFse 分析组间菌群差异，壳糖平中高剂量组拟杆菌群占主导，且所占比例较高；相关研究表明，毛螺科菌(Lachnospiraceae)与肠道炎症程度正相关；人类二型糖尿病和T2DM模型小鼠的发病可能与Lachnospi raceae相关。据报告显示，Desulfovibrionaceae是一种硫酸盐还原菌，能够产生毒素并破坏粘膜屏障与炎症密切相关；Prevotellaceae是一种与肠道的通透性相关的黏蛋白降解菌，在抵御病原菌方面发挥重要作用；肠道中的Bacteroidales_S24-7_group是丁酸盐产生菌，丁酸盐是肠道上皮的能量提供源，并有助于肠道对营养物质的消化吸收，从而提高机体的肠道免疫力。
本研究结果表明，给予灌服壳糖平可以改善T2DM小鼠肠道炎症程度，促进肠道有益菌的生长而抑制致病菌的增值，从而使得肠道免疫力增强，同时使得宿主整体内环境协调稳定。并且灌服壳糖平与阳性药(Acar)相比，壳糖平的作用更温和，对肠道有益菌的定植更高效。
3.2.4不同分组之间OTU物种差异和PCA分析
如图8，根据OTU丰度信息开展韦恩图分析，7组样本含有的特有的OTU个数分别为326(NFD组)、223(T2DM组)、237(Ac ar组)、118(COS组)、197(LKTP组)、142(MKTP组)、18 1(HKTP组)。其中有556个OTU是7组共有。组间PCoA分析结果说明，小鼠肠道样品越相似，其反映在PCoA图中的距离越近，对于不同环境中的样品可能会出现单独聚类分布的情况，如图9表明，表现在PCoA坐标上的距离，Acar组距离其他各组都比较远，说明A car组的小鼠肠道菌群组成及结构与其他组差异显著；T2DM组、CO S组和NFD组菌落组成最相近，差异不显著，LKTP组、MKTP组和 HKTP组菌落组成较为相似，差异不显著。
以上内容是结合具体的优选实施方式对本发明所作的进一步详细说明，不能认定本发明的具体实施只局限于这些说明。对于本发明所属技术领域的普通技术人员来说，在不脱离本发明构思的前提下，还可以做出若干简单推演或替换，都应当视为属于本发明的保护范围。
Claims (3)
1.一种辅助治疗二型糖尿病的壳寡糖复合固体饮料，其特征在于，所述壳聚糖复合固体饮料包括壳寡糖、水苏糖、燕麦β-葡聚糖、白芸豆提取物和亚麻籽油微囊粉。
2.根据权利要求1所述的辅助治疗二型糖尿病的壳寡糖复合固体饮料，其特征在于，按重量百分数计，所述壳聚糖复合固体饮料中各组分的含量如下：壳寡糖5～30％；水苏糖2～18％；燕麦β-葡聚糖5～20％；白芸豆提取物16～38％；亚麻籽油微囊粉10～20％。
3.根据权利要求2所述的辅助治疗二型糖尿病的壳寡糖复合固体饮料，其特征在于，按重量百分数计，壳寡糖28％；水苏糖18％；燕麦β-葡聚糖18％；白芸豆提取物16％；亚麻籽油微囊粉20％。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911303224.9A CN110771769A (zh) | 2019-12-17 | 2019-12-17 | 一种辅助治疗二型糖尿病的壳寡糖复合固体饮料 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201911303224.9A CN110771769A (zh) | 2019-12-17 | 2019-12-17 | 一种辅助治疗二型糖尿病的壳寡糖复合固体饮料 |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110771769A true CN110771769A (zh) | 2020-02-11 |
Family
ID=69394633
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201911303224.9A Pending CN110771769A (zh) | 2019-12-17 | 2019-12-17 | 一种辅助治疗二型糖尿病的壳寡糖复合固体饮料 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110771769A (zh) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112450385A (zh) * | 2020-12-02 | 2021-03-09 | 宣城柏维力生物工程有限公司 | 一种白芸豆壳糖直饮粉制备工艺 |
-
2019
- 2019-12-17 CN CN201911303224.9A patent/CN110771769A/zh active Pending
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN112450385A (zh) * | 2020-12-02 | 2021-03-09 | 宣城柏维力生物工程有限公司 | 一种白芸豆壳糖直饮粉制备工艺 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| Zhang et al. | Effects of Auricularia auricula and its polysaccharide on diet-induced hyperlipidemia rats by modulating gut microbiota | |
| JP2022000040A (ja) | 代謝障害を処置するための低温殺菌アッカーマンシアの使用 | |
| Ran et al. | Sea buckthorn (Hippophae rhamnoides L.) fermentation liquid protects against alcoholic liver disease linked to regulation of liver metabolome and the abundance of gut microbiota | |
| Raheja et al. | Lactobacillus acidophilus stimulates the expression of SLC26A3 via a transcriptional mechanism | |
| CN110150669B (zh) | 一种适于糖尿病患者食用的益生菌组合物及其应用 | |
| CN109689077B (zh) | 新用途及方法 | |
| TWI667036B (zh) | 植物萃取物於調控ｐｄｇｆｃ、ｆｇｆ２、ｉｇｆ１ｒ、ｐｔｇｉｓ、ｎｏｓ３、ｅｄｎ１、ｐｌａｔ、ｐｒｏｃ、ｖｗｆ、ｆ３、ｓｅｒｐｉｎｅ１、ｉｌ－８、ｉｃａｍ１、ｖｃａｍ１及ｃａｓｐ８基因的用途 | |
| Pang et al. | Lactobacillus rhamnosus from human breast milk ameliorates ulcerative colitis in mice via gut microbiota modulation | |
| Ma et al. | Oyster (Crassostrea gigas) polysaccharide ameliorates obesity in association with modulation of lipid metabolism and gut microbiota in high-fat diet fed mice | |
| KR102282490B1 (ko) | 패칼리박테리움 프라우스니찌 유래 나노소포 및 이의 용도 | |
| Lu et al. | Eurotium cristatum, a probiotic fungus from Fuzhuan brick tea, and its polysaccharides ameliorated DSS-induced ulcerative colitis in mice by modulating the gut microbiota | |
| CN113197921B (zh) | 乳双歧杆菌MN-Gup及其菌剂在治疗2型糖尿病中的应用 | |
| US11898156B2 (en) | Nano-vesicles derived from genus Morganella bacteria and use thereof | |
| Wan et al. | Dicaffeoylquinic acids from Ilex kudingcha attenuate dextran sulfate sodium-induced colitis in C57BL/6 mice in association with the modulation of gut microbiota | |
| Gallotti et al. | Effects of dietary fibre intake in chemotherapy-induced mucositis in murine model | |
| Wang et al. | Methamphetamine induces intestinal injury by altering gut microbiota and promoting inflammation in mice | |
| Chen et al. | Fructus mori polysaccharide alleviates diabetic symptoms by regulating intestinal microbiota and intestinal barrier against TLR4/NF-κB pathway | |
| Lu et al. | Effects of exopolysaccharides from Antrodia cinnamomea on inflammation and intestinal microbiota disturbance induced by antibiotics in mice | |
| CN110771769A (zh) | 一种辅助治疗二型糖尿病的壳寡糖复合固体饮料 | |
| WO2019109551A1 (zh) | 一种含有铝碳酸镁的药物组合物及制药用途 | |
| KR102118996B1 (ko) | 베일로넬라 속 세균 유래 나노소포 및 이의 용도 | |
| Gomes et al. | Cooked common bean flour, but not its protein hydrolysate, has the potential to improve gut microbiota composition and function in BALB/c mice fed a high-fat diet added with 6-propyl-2-thiouracil | |
| KR102122903B1 (ko) | 블라우티아 속 세균 유래 나노소포 및 이의 용도 | |
| Leng et al. | Protective Effect of Ganoderma lucidum Spore Powder on Acute Liver Injury in Mice and its Regulation of Gut Microbiota | |
| CN111019010B (zh) | Nigella sativa种籽多糖、提取方法和制备治疗2型糖尿病药物的应用 |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| WD01 | Invention patent application deemed withdrawn after publication |
Application publication date: 20200211 |
|
| WD01 | Invention patent application deemed withdrawn after publication |