CN116376740A - Bifidobacterium longum with blood sugar reducing effect and application thereof - Google Patents
Bifidobacterium longum with blood sugar reducing effect and application thereof Download PDFInfo
- Publication number
- CN116376740A CN116376740A CN202211588301.1A CN202211588301A CN116376740A CN 116376740 A CN116376740 A CN 116376740A CN 202211588301 A CN202211588301 A CN 202211588301A CN 116376740 A CN116376740 A CN 116376740A
- Authority
- CN
- China
- Prior art keywords
- bifidobacterium longum
- longum
- composition
- group
- effect
- 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
- 210000004369 blood Anatomy 0.000 title claims abstract description 73
- 239000008280 blood Substances 0.000 title claims abstract description 73
- 241001608472 Bifidobacterium longum Species 0.000 title claims abstract description 72
- 229940009291 bifidobacterium longum Drugs 0.000 title claims abstract description 72
- 230000001603 reducing effect Effects 0.000 title claims abstract description 19
- 230000000694 effects Effects 0.000 claims abstract description 18
- 150000004666 short chain fatty acids Chemical class 0.000 claims abstract description 18
- 241000894006 Bacteria Species 0.000 claims abstract description 17
- 230000009286 beneficial effect Effects 0.000 claims abstract description 13
- 244000005700 microbiome Species 0.000 claims abstract description 10
- 238000000034 method Methods 0.000 claims abstract description 6
- 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 claims description 66
- 239000008103 glucose Substances 0.000 claims description 66
- 230000001580 bacterial effect Effects 0.000 claims description 31
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 claims description 27
- 239000000203 mixture Substances 0.000 claims description 22
- 210000002966 serum Anatomy 0.000 claims description 21
- 230000000968 intestinal effect Effects 0.000 claims description 16
- 239000000843 powder Substances 0.000 claims description 12
- 241000186000 Bifidobacterium Species 0.000 claims description 11
- 239000000243 solution Substances 0.000 claims description 11
- XBDQKXXYIPTUBI-UHFFFAOYSA-N dimethylselenoniopropionate Natural products CCC(O)=O XBDQKXXYIPTUBI-UHFFFAOYSA-N 0.000 claims description 10
- 238000002360 preparation method Methods 0.000 claims description 10
- 235000021391 short chain fatty acids Nutrition 0.000 claims description 10
- 230000014509 gene expression Effects 0.000 claims description 9
- 239000007788 liquid Substances 0.000 claims description 9
- 108090000045 G-Protein-Coupled Receptors Proteins 0.000 claims description 8
- 230000002218 hypoglycaemic effect Effects 0.000 claims description 7
- 239000003795 chemical substances by application Substances 0.000 claims description 6
- 235000019260 propionic acid Nutrition 0.000 claims description 5
- IUVKMZGDUIUOCP-BTNSXGMBSA-N quinbolone Chemical compound O([C@H]1CC[C@H]2[C@H]3[C@@H]([C@]4(C=CC(=O)C=C4CC3)C)CC[C@@]21C)C1=CCCC1 IUVKMZGDUIUOCP-BTNSXGMBSA-N 0.000 claims description 5
- 102100025101 GATA-type zinc finger protein 1 Human genes 0.000 claims description 4
- 101710198884 GATA-type zinc finger protein 1 Proteins 0.000 claims description 4
- DTHNMHAUYICORS-KTKZVXAJSA-N Glucagon-like peptide 1 Chemical compound C([C@@H](C(=O)N[C@@H]([C@@H](C)CC)C(=O)N[C@@H](C)C(=O)N[C@@H](CC=1C2=CC=CC=C2NC=1)C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](C(C)C)C(=O)N[C@@H](CCCCN)C(=O)NCC(=O)N[C@@H](CCCNC(N)=N)C(N)=O)NC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CCCCN)NC(=O)[C@H](C)NC(=O)[C@H](C)NC(=O)[C@H](CCC(N)=O)NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CC=1C=CC(O)=CC=1)NC(=O)[C@H](CO)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC(O)=O)NC(=O)[C@H](CO)NC(=O)[C@@H](NC(=O)[C@H](CC=1C=CC=CC=1)NC(=O)[C@@H](NC(=O)CNC(=O)[C@H](CCC(O)=O)NC(=O)[C@H](C)NC(=O)[C@@H](N)CC=1N=CNC=1)[C@@H](C)O)[C@@H](C)O)C(C)C)C1=CC=CC=C1 DTHNMHAUYICORS-KTKZVXAJSA-N 0.000 claims description 4
- 239000003629 gastrointestinal hormone Substances 0.000 claims description 4
- 238000012545 processing Methods 0.000 claims description 4
- 230000028327 secretion Effects 0.000 claims description 4
- 239000002775 capsule Substances 0.000 claims description 3
- 238000012258 culturing Methods 0.000 claims description 3
- 239000008187 granular material Substances 0.000 claims description 3
- 239000002245 particle Substances 0.000 claims description 3
- 239000006187 pill Substances 0.000 claims description 3
- 239000006041 probiotic Substances 0.000 claims description 3
- 235000018291 probiotics Nutrition 0.000 claims description 3
- 230000001737 promoting effect Effects 0.000 claims description 3
- 239000003826 tablet Substances 0.000 claims description 3
- 230000000529 probiotic effect Effects 0.000 claims description 2
- 238000004519 manufacturing process Methods 0.000 claims 3
- 230000003028 elevating effect Effects 0.000 claims 1
- 241001465754 Metazoa Species 0.000 abstract description 21
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 14
- 210000003608 fece Anatomy 0.000 abstract description 9
- 231100000331 toxic Toxicity 0.000 abstract description 8
- 230000002588 toxic effect Effects 0.000 abstract description 8
- 208000030159 metabolic disease Diseases 0.000 abstract description 7
- 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 abstract description 6
- 229930186217 Glycolipid Natural products 0.000 abstract description 6
- 108090000623 proteins and genes Proteins 0.000 abstract description 6
- 238000011156 evaluation Methods 0.000 abstract description 5
- 239000003242 anti bacterial agent Substances 0.000 abstract description 4
- 229940088710 antibiotic agent Drugs 0.000 abstract description 4
- 229940126672 traditional medicines Drugs 0.000 abstract description 3
- 241000700159 Rattus Species 0.000 description 36
- NOESYZHRGYRDHS-UHFFFAOYSA-N insulin Chemical compound N1C(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(NC(=O)CN)C(C)CC)CSSCC(C(NC(CO)C(=O)NC(CC(C)C)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CCC(N)=O)C(=O)NC(CC(C)C)C(=O)NC(CCC(O)=O)C(=O)NC(CC(N)=O)C(=O)NC(CC=2C=CC(O)=CC=2)C(=O)NC(CSSCC(NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2C=CC(O)=CC=2)NC(=O)C(CC(C)C)NC(=O)C(C)NC(=O)C(CCC(O)=O)NC(=O)C(C(C)C)NC(=O)C(CC(C)C)NC(=O)C(CC=2NC=NC=2)NC(=O)C(CO)NC(=O)CNC2=O)C(=O)NCC(=O)NC(CCC(O)=O)C(=O)NC(CCCNC(N)=N)C(=O)NCC(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC=CC=3)C(=O)NC(CC=3C=CC(O)=CC=3)C(=O)NC(C(C)O)C(=O)N3C(CCC3)C(=O)NC(CCCCN)C(=O)NC(C)C(O)=O)C(=O)NC(CC(N)=O)C(O)=O)=O)NC(=O)C(C(C)CC)NC(=O)C(CO)NC(=O)C(C(C)O)NC(=O)C1CSSCC2NC(=O)C(CC(C)C)NC(=O)C(NC(=O)C(CCC(N)=O)NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(N)CC=1C=CC=CC=1)C(C)C)CC1=CN=CN1 NOESYZHRGYRDHS-UHFFFAOYSA-N 0.000 description 18
- 241000699666 Mus <mouse, genus> Species 0.000 description 16
- 208000001072 type 2 diabetes mellitus Diseases 0.000 description 15
- 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 14
- 239000000725 suspension Substances 0.000 description 14
- 241000699670 Mus sp. Species 0.000 description 13
- 238000012360 testing method Methods 0.000 description 12
- 206010012601 diabetes mellitus Diseases 0.000 description 11
- 230000002496 gastric effect Effects 0.000 description 11
- 230000037396 body weight Effects 0.000 description 10
- XZWYZXLIPXDOLR-UHFFFAOYSA-N metformin Chemical compound CN(C)C(=N)NC(N)=N XZWYZXLIPXDOLR-UHFFFAOYSA-N 0.000 description 10
- 102000004877 Insulin Human genes 0.000 description 9
- 108090001061 Insulin Proteins 0.000 description 9
- 206010022489 Insulin Resistance Diseases 0.000 description 9
- 229940125396 insulin Drugs 0.000 description 9
- 235000011054 acetic acid Nutrition 0.000 description 8
- 239000002504 physiological saline solution Substances 0.000 description 8
- 238000004458 analytical method Methods 0.000 description 7
- 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 7
- 238000002474 experimental method Methods 0.000 description 7
- 229960003105 metformin Drugs 0.000 description 7
- 235000005911 diet Nutrition 0.000 description 6
- 230000037213 diet Effects 0.000 description 6
- 239000003814 drug Substances 0.000 description 6
- 239000001963 growth medium Substances 0.000 description 6
- 150000002632 lipids Chemical class 0.000 description 6
- 101000890668 Homo sapiens Free fatty acid receptor 2 Proteins 0.000 description 5
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 5
- 208000035475 disorder Diseases 0.000 description 5
- 229940079593 drug Drugs 0.000 description 5
- 239000007928 intraperitoneal injection Substances 0.000 description 5
- 235000021590 normal diet Nutrition 0.000 description 5
- 238000004393 prognosis Methods 0.000 description 5
- 210000002784 stomach Anatomy 0.000 description 5
- 108020004465 16S ribosomal RNA Proteins 0.000 description 4
- 102000003688 G-Protein-Coupled Receptors Human genes 0.000 description 4
- 208000017170 Lipid metabolism disease Diseases 0.000 description 4
- 208000018914 glucose metabolism disease Diseases 0.000 description 4
- 238000012423 maintenance Methods 0.000 description 4
- 239000000902 placebo Substances 0.000 description 4
- 229940068196 placebo Drugs 0.000 description 4
- 239000000047 product Substances 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- 239000011550 stock solution Substances 0.000 description 4
- 239000006228 supernatant Substances 0.000 description 4
- 150000003626 triacylglycerols Chemical class 0.000 description 4
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 4
- 108020004414 DNA Proteins 0.000 description 3
- 102100040136 Free fatty acid receptor 3 Human genes 0.000 description 3
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 101000890662 Homo sapiens Free fatty acid receptor 3 Proteins 0.000 description 3
- 238000005119 centrifugation Methods 0.000 description 3
- 230000008859 change Effects 0.000 description 3
- 235000012000 cholesterol Nutrition 0.000 description 3
- 238000001514 detection method Methods 0.000 description 3
- 201000001421 hyperglycemia Diseases 0.000 description 3
- 239000007924 injection Substances 0.000 description 3
- 238000002347 injection Methods 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 238000003305 oral gavage Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- QIJRTFXNRTXDIP-UHFFFAOYSA-N (1-carboxy-2-sulfanylethyl)azanium;chloride;hydrate Chemical compound O.Cl.SCC(N)C(O)=O QIJRTFXNRTXDIP-UHFFFAOYSA-N 0.000 description 2
- 241000186361 Actinobacteria <class> Species 0.000 description 2
- 208000013016 Hypoglycemia Diseases 0.000 description 2
- 230000007059 acute toxicity Effects 0.000 description 2
- 231100000403 acute toxicity Toxicity 0.000 description 2
- 210000004027 cell Anatomy 0.000 description 2
- 230000001684 chronic effect Effects 0.000 description 2
- 235000015140 cultured milk Nutrition 0.000 description 2
- 229960001305 cysteine hydrochloride Drugs 0.000 description 2
- 230000002550 fecal effect Effects 0.000 description 2
- 238000011049 filling Methods 0.000 description 2
- 235000013305 food Nutrition 0.000 description 2
- 230000001404 mediated effect Effects 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 239000006872 mrs medium Substances 0.000 description 2
- 210000000496 pancreas Anatomy 0.000 description 2
- 230000007918 pathogenicity Effects 0.000 description 2
- 238000011552 rat model Methods 0.000 description 2
- 239000002994 raw material Substances 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 239000008223 sterile water Substances 0.000 description 2
- 239000004575 stone 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
- 239000000126 substance Substances 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 1
- 241000606125 Bacteroides Species 0.000 description 1
- 241001134770 Bifidobacterium animalis Species 0.000 description 1
- 241000186015 Bifidobacterium longum subsp. infantis Species 0.000 description 1
- 208000031648 Body Weight Changes Diseases 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 235000019733 Fish meal Nutrition 0.000 description 1
- 208000003790 Foot Ulcer Diseases 0.000 description 1
- 102100040133 Free fatty acid receptor 2 Human genes 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 229930182566 Gentamicin Natural products 0.000 description 1
- CEAZRRDELHUEMR-URQXQFDESA-N Gentamicin Chemical compound O1[C@H](C(C)NC)CC[C@@H](N)[C@H]1O[C@H]1[C@H](O)[C@@H](O[C@@H]2[C@@H]([C@@H](NC)[C@@](C)(O)CO2)O)[C@H](N)C[C@@H]1N CEAZRRDELHUEMR-URQXQFDESA-N 0.000 description 1
- 240000008415 Lactuca sativa Species 0.000 description 1
- 240000004658 Medicago sativa Species 0.000 description 1
- 235000017587 Medicago sativa ssp. sativa Nutrition 0.000 description 1
- 241001529936 Murinae Species 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 208000008589 Obesity Diseases 0.000 description 1
- 208000017442 Retinal disease Diseases 0.000 description 1
- 206010038923 Retinopathy Diseases 0.000 description 1
- 235000019764 Soybean Meal Nutrition 0.000 description 1
- 229930006000 Sucrose Natural products 0.000 description 1
- 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 1
- 241001052560 Thallis Species 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum Nutrition 0.000 description 1
- 206010067584 Type 1 diabetes mellitus Diseases 0.000 description 1
- 108010059993 Vancomycin Proteins 0.000 description 1
- 208000000260 Warts Diseases 0.000 description 1
- 235000019752 Wheat Middilings Nutrition 0.000 description 1
- 240000008042 Zea mays Species 0.000 description 1
- 235000005824 Zea mays ssp. parviglumis Nutrition 0.000 description 1
- 235000002017 Zea mays subsp mays Nutrition 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 229960000723 ampicillin Drugs 0.000 description 1
- AVKUERGKIZMTKX-NJBDSQKTSA-N ampicillin Chemical compound C1([C@@H](N)C(=O)N[C@H]2[C@H]3SC([C@@H](N3C2=O)C(O)=O)(C)C)=CC=CC=C1 AVKUERGKIZMTKX-NJBDSQKTSA-N 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 229940118852 bifidobacterium animalis Drugs 0.000 description 1
- 229940004120 bifidobacterium infantis Drugs 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000004579 body weight change Effects 0.000 description 1
- 210000000988 bone and bone Anatomy 0.000 description 1
- NKWPZUCBCARRDP-UHFFFAOYSA-L calcium bicarbonate Chemical compound [Ca+2].OC([O-])=O.OC([O-])=O NKWPZUCBCARRDP-UHFFFAOYSA-L 0.000 description 1
- 229910000020 calcium bicarbonate Inorganic materials 0.000 description 1
- 239000005018 casein Substances 0.000 description 1
- BECPQYXYKAMYBN-UHFFFAOYSA-N casein, tech. Chemical compound NCCCCC(C(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(CC(C)C)N=C(O)C(CCC(O)=O)N=C(O)C(CC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(C(C)O)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=N)N=C(O)C(CCC(O)=O)N=C(O)C(CCC(O)=O)N=C(O)C(COP(O)(O)=O)N=C(O)C(CCC(O)=N)N=C(O)C(N)CC1=CC=CC=C1 BECPQYXYKAMYBN-UHFFFAOYSA-N 0.000 description 1
- 235000021240 caseins Nutrition 0.000 description 1
- 229960005091 chloramphenicol Drugs 0.000 description 1
- WIIZWVCIJKGZOK-RKDXNWHRSA-N chloramphenicol Chemical compound ClC(Cl)C(=O)N[C@H](CO)[C@H](O)C1=CC=C([N+]([O-])=O)C=C1 WIIZWVCIJKGZOK-RKDXNWHRSA-N 0.000 description 1
- 208000037976 chronic inflammation Diseases 0.000 description 1
- 230000006020 chronic inflammation Effects 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 238000000576 coating method Methods 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 238000012937 correction Methods 0.000 description 1
- 238000011697 diabetes animal model Methods 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 208000016097 disease of metabolism Diseases 0.000 description 1
- 235000013345 egg yolk Nutrition 0.000 description 1
- 210000002969 egg yolk Anatomy 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000004467 fishmeal Substances 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 238000012268 genome sequencing Methods 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- 208000004104 gestational diabetes Diseases 0.000 description 1
- 230000004153 glucose metabolism Effects 0.000 description 1
- 238000007446 glucose tolerance test Methods 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 235000009200 high fat diet Nutrition 0.000 description 1
- 235000021069 high fat-high sugar diet Nutrition 0.000 description 1
- 238000013225 high-fat diet-induced obese mouse model Methods 0.000 description 1
- 230000003345 hyperglycaemic effect Effects 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- XFIKMPRBSORKQP-JATHGWPISA-M lithium;9-[(e)-4-[(2s,3r,4r,5s)-3,4-dihydroxy-5-[[(2s,3s)-3-[(2s,3s)-3-hydroxybutan-2-yl]oxiran-2-yl]methyl]oxan-2-yl]-3-methylbut-2-enoyl]oxynonanoate Chemical class [Li+].C[C@H](O)[C@H](C)[C@@H]1O[C@H]1C[C@@H]1[C@@H](O)[C@@H](O)[C@H](C\C(C)=C\C(=O)OCCCCCCCCC([O-])=O)OC1 XFIKMPRBSORKQP-JATHGWPISA-M 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000008176 lyophilized powder Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000002609 medium Substances 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 210000000214 mouth Anatomy 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 231100000956 nontoxicity Toxicity 0.000 description 1
- 150000007523 nucleic acids Chemical group 0.000 description 1
- 235000020824 obesity Nutrition 0.000 description 1
- 210000004923 pancreatic tissue Anatomy 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 238000004321 preservation Methods 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 239000003223 protective agent Substances 0.000 description 1
- 235000018102 proteins Nutrition 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 238000003753 real-time PCR Methods 0.000 description 1
- 230000001105 regulatory effect Effects 0.000 description 1
- 235000012045 salad Nutrition 0.000 description 1
- 238000007790 scraping Methods 0.000 description 1
- 238000012216 screening Methods 0.000 description 1
- 239000006152 selective media Substances 0.000 description 1
- 230000035945 sensitivity Effects 0.000 description 1
- 201000010153 skin papilloma Diseases 0.000 description 1
- NRHMKIHPTBHXPF-TUJRSCDTSA-M sodium cholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 NRHMKIHPTBHXPF-TUJRSCDTSA-M 0.000 description 1
- 239000004455 soybean meal Substances 0.000 description 1
- 241000894007 species Species 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 230000004936 stimulating effect Effects 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 239000005720 sucrose Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000009885 systemic effect Effects 0.000 description 1
- 230000001225 therapeutic effect Effects 0.000 description 1
- 230000009466 transformation Effects 0.000 description 1
- 210000002700 urine Anatomy 0.000 description 1
- 210000001215 vagina Anatomy 0.000 description 1
- 229960003165 vancomycin Drugs 0.000 description 1
- MYPYJXKWCTUITO-LYRMYLQWSA-N vancomycin Chemical compound O([C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=C2C=C3C=C1OC1=CC=C(C=C1Cl)[C@@H](O)[C@H](C(N[C@@H](CC(N)=O)C(=O)N[C@H]3C(=O)N[C@H]1C(=O)N[C@H](C(N[C@@H](C3=CC(O)=CC(O)=C3C=3C(O)=CC=C1C=3)C(O)=O)=O)[C@H](O)C1=CC=C(C(=C1)Cl)O2)=O)NC(=O)[C@@H](CC(C)C)NC)[C@H]1C[C@](C)(N)[C@H](O)[C@H](C)O1 MYPYJXKWCTUITO-LYRMYLQWSA-N 0.000 description 1
- MYPYJXKWCTUITO-UHFFFAOYSA-N vancomycin Natural products O1C(C(=C2)Cl)=CC=C2C(O)C(C(NC(C2=CC(O)=CC(O)=C2C=2C(O)=CC=C3C=2)C(O)=O)=O)NC(=O)C3NC(=O)C2NC(=O)C(CC(N)=O)NC(=O)C(NC(=O)C(CC(C)C)NC)C(O)C(C=C3Cl)=CC=C3OC3=CC2=CC1=C3OC1OC(CO)C(O)C(O)C1OC1CC(C)(N)C(O)C(C)O1 MYPYJXKWCTUITO-UHFFFAOYSA-N 0.000 description 1
- 230000001018 virulence Effects 0.000 description 1
- 239000011782 vitamin Substances 0.000 description 1
- 229940088594 vitamin Drugs 0.000 description 1
- 229930003231 vitamin Natural products 0.000 description 1
- 235000013343 vitamin Nutrition 0.000 description 1
- 238000012070 whole genome sequencing analysis Methods 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
- C12N1/205—Bacterial isolates
-
- 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
- A61K35/745—Bifidobacteria
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/08—Drugs for disorders of the metabolism for glucose homeostasis
- A61P3/10—Drugs for disorders of the metabolism for glucose homeostasis for hyperglycaemia, e.g. antidiabetics
-
- 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
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Abstract
The invention relates to the field of microorganisms, and discloses bifidobacterium longum with blood sugar reducing effect and application thereof. The bifidobacterium longum is named WH2270, and the microorganism classification is named bifidobacterium longum subspecies longumBifidobacterium longum subsp. LongumThe method comprises the steps of carrying out a first treatment on the surface of the Has been deposited at the microorganism strain collection of Guangdong province at 10.18 of 2022, with the deposit address of Hirschner No. 100 in Guangzhou City and the deposit number of GDMCC No:62900. the bifidobacterium longum subspecies longum of the invention can improve the abundance of beneficial bacteria in the intestinal tract, can improve the content of short chain fatty acid in the intestinal tract, has excellent blood sugar reducing effect,the strain is separated from the feces of healthy infants, has no toxic genes, has no toxic or side effects in animal evaluation, is sensitive to various antibiotics, and has certain advantages compared with the traditional medicines for treating glycolipid metabolic disorders at present.
Description
Technical Field
The invention relates to the field of microorganisms, in particular to bifidobacterium longum with blood sugar reducing effect and application thereof.
Background
Diabetes is a chronic metabolic disease characterized by hyperglycemia, also known as an intangible killer, because hyperglycemia is harmful to the human body, but is difficult to detect. Long-term hyperglycemic symptoms can cause a range of chronic complications, such as retinopathy, nephropathy, foot ulcers, and the like. Diabetes is largely divided into 3 types: type 1, type 2 and gestational diabetes mellitus, wherein Type 2diabetes mellitus (t 2 dm) accounts for about 90% of the total number. Therefore, the preventive and therapeutic measures for developing type 2diabetes are not known.
Intestinal flora plays an important role in the pathogenesis and metabolic disorders of type 2 diabetes. Excessive intake of high-fat and high-sugar foods can disrupt normal intestinal flora, thereby inducing systemic, low-grade chronic inflammation and causing metabolic diseases such as obesity and type 2 diabetes. Studies have found that the level of harmful bacteria in the feces of type 2 diabetics is elevated compared to healthy people, such as firmicutes, actinomycetes and escherichia coli; the level of beneficial bacteria is reduced, for example, bacteroides and bifidobacteria. Intestinal flora is one of the targets for intervention in type 2 diabetes.
Bifidobacterium (bifidobacteria) is a gram-positive, motionless, rod-shaped, sometimes bifurcated at one end, strictly anaerobic bacterium which is widely found in the digestive tract, vagina, oral cavity and other habitats of humans and animals, and is an important component member of the intestinal flora of humans and animals. Animal experiments have found that multiple species of bifidobacterium (bifidobacterium longum, bifidobacterium infantis, bifidobacterium animalis, etc.) can increase host glucose tolerance. Bifidobacterium longum is a main type of bifidobacterium in the intestinal tract of adults, has good colonization ability in the intestinal tract of hosts, and has been reported to have hypoglycemic effects. Patent publication No. CN114774335A reports a bifidobacterium longum 070103 (GDMCC No. 62526), and the fermented milk prepared by fermenting the strain is applied to a mouse model with glycolipid disorder caused by gastric lavage high-fat high-sugar diet, so that the fermented milk has the efficacy of reducing blood sugar and blood fat, but the research on the blood sugar reducing effect of the strain per se is lacking; patent publication No. CN113293113A reports a bifidobacterium longum MI-186 (CGMCC No. 21586), and a gastric lavage high fat diet induced obese mouse model is found to have the effects of reducing blood fat and blood sugar, but the strain lacks the blood sugar reducing effect research on a diabetes animal model and the influence on the blood sugar of normal test animals; patent publication No. CN113755409A reports that a bifidobacterium longum NSP008 (GDMCC No: 61889), a gastric lavage high fat diet induced insulin resistant mouse model, can improve glucose tolerance and reduce blood glucose in mice, but the strain is isolated from fecal samples of type 2 diabetics and is at risk of strain origin.
Disclosure of Invention
In order to solve the technical problems, the invention provides bifidobacterium longum with blood sugar reducing effect and application thereof. The bifidobacterium longum subspecies longum disclosed by the invention can be used for improving the abundance of beneficial bacteria in intestinal tracts, improving the content of short-chain fatty acids in the intestinal tracts, has excellent blood sugar reducing effect, is separated from feces of healthy infants, has no toxic genes, is free from toxic or side effects in animal evaluation, is sensitive to various antibiotics, and has certain advantages compared with the traditional medicines for treating glycolipid metabolic disorders at present.
The specific technical scheme of the invention is as follows:
in a first aspect, the invention provides a bifidobacterium longum with hypoglycemic effect, named WHH2270, and the microorganism classification named bifidobacterium longum subsp. Bifidobacterium longum subsp. Has been deposited at the microorganism strain collection of Guangdong province at 10.18 of 2022, with the deposit address of Hirschner No. 100 in Guangzhou City and the deposit number of GDMCC No:62900.
in a second aspect, the present invention provides the use of a bifidobacterium longum as described above in the preparation of a composition having the effect of improving the abundance of intestinal beneficial bacteria.
Further, the beneficial bacteria include bifidobacteria and AKK bacteria.
The invention provides application of bifidobacterium longum in preparing a composition with the effect of improving the content of short-chain fatty acids in intestinal tracts and serum.
Further, the short chain fatty acids are acetic acid and propionic acid.
The invention provides application of bifidobacterium longum in preparing a composition with the function of improving the expression level of a G protein coupled receptor gene.
The invention provides application of bifidobacterium longum in preparing a composition with an effect of promoting secretion of intestinal hormone GLP-1 and/or PYY.
The invention provides application of bifidobacterium longum in preparation of a composition with blood sugar reducing effect.
Preferably, the composition is an oral composition.
Further preferably, the composition is in the form of a powder, tablet, capsule, granule, pill or solution.
In a third aspect, the invention provides a probiotic bacterial agent, which is bacterial liquid obtained after culturing bifidobacterium longum or a cryogenic particle or dry powder bacterial agent obtained by further processing the bacterial liquid.
Compared with the prior art, the invention has the beneficial effects that:
(1) The bifidobacterium longum subspecies of the invention have the function of reducing blood sugar for animals with glucose-lipid disorder of insulin resistance, and can not influence normal blood sugar for healthy animals. Namely, the bifidobacterium longum subspecies longum has the effect of reducing blood sugar only for patients with hyperglycemia after being taken, does not have negative influence on blood sugar of normal people, and does not cause the occurrence of hypoglycemia. Therefore, the preparation method has no side effect and wider application field, and can be applied to not only the medicine field but also the food or health care product field and the like.
(2) The bifidobacterium longum subspecies longum can regulate the disturbed intestinal flora and improve the abundance of beneficial bacteria in intestinal tracts.
(3) The bifidobacterium longum subspecies longum disclosed by the invention can be used for improving the content of short-chain fatty acid in intestinal tracts, improving the content of acetic acid in serum, stimulating the expression of G protein coupled receptors of the intestinal tracts and pancreas-mediated short-chain fatty acid, and reducing blood sugar.
(4) The bifidobacterium longum subspecies are derived from the feces of healthy infants, and the whole genome sequencing analysis shows that the strain has no toxic genes, has no toxic or side effects in animal evaluation, is sensitive to various antibiotics, and has certain advantages compared with the traditional medicines for treating glycolipid metabolic disorders at present.
Drawings
Figure 1 shows the body weight change of each group of rats. There was no significant difference between groups;
FIG. 2 shows blood glucose levels of 0.5h and 2h after fasting blood glucose and oral glucose before treatment of each group of rats. There was no significant difference between groups;
FIG. 3 is a graph showing fasting blood glucose, 0.5h after oral glucose, 2h after oral glucose, and glucose tolerance curves for each group of rats after 8 weeks of sample treatment;
FIG. 4 shows serum TC, TG of each group of rats after 8 weeks of sample treatment;
FIG. 5 is an analysis of the intestinal flora of rats in the bifidobacterium longum WH 2270 intervention group and model and placebo groups;
FIG. 6 shows the acetic acid content in the intestinal short-chain fatty acid and serum of rats in the bifidobacterium longum WH 2270 intervention group and model and placebo groups;
FIG. 7 shows the gene expression of the G protein-coupled receptors GPR41 and GPR43 in intestinal and pancreatic tissue of rats in the bifidobacterium longum WH 2270 intervention group and model and placebo groups.
Detailed Description
The invention is further described below with reference to examples.
General examples
Bifidobacterium longum with hypoglycemic effect is named WH2270, and the microorganism classification is named as Bifidobacterium longum subsp. Has been deposited at the microorganism strain collection of Guangdong province at 10.18 of 2022, with the deposit address of Hirschner No. 100 in Guangzhou City and the deposit number of GDMCC No:62900.
the application of the bifidobacterium longum in preparing a composition with the effect of improving the abundance of beneficial intestinal bacteria. Further, the beneficial bacteria include bifidobacteria and AKK bacteria.
The invention provides application of bifidobacterium longum in preparing a composition with the effect of improving the content of short-chain fatty acids in intestinal tracts and serum. Further, the short chain fatty acids are acetic acid and propionic acid.
The invention provides application of bifidobacterium longum in preparing a composition with the function of improving the expression level of a G protein coupled receptor gene.
The invention provides application of bifidobacterium longum in preparing a composition with an effect of promoting secretion of intestinal hormone GLP-1 and/or PYY.
The invention provides application of bifidobacterium longum in preparation of a composition with blood sugar reducing effect.
Preferably, the composition is an oral composition. Further preferably, the composition is in the form of a powder, tablet, capsule, granule, pill or solution.
The probiotics bacterial agent is bacterial liquid obtained after the bifidobacterium longum is cultured, or cryogenic particles or dry powder bacterial agent obtained by further processing the bacterial liquid.
Example 1: isolation, identification, preservation and Property measurement of strains
1. 16S rDNA identification
Adding 10g of a fecal sample of a healthy infant into 90mL of sterile physiological saline, shaking and uniformly mixing, carrying out gradient dilution, coating an MRS and mupirocin lithium salt selective medium plate, placing the plate at 37 ℃, carrying out anaerobic culture for 48 hours, picking single colony for purification, inoculating the pure single colony into 10mL of MRS Broth medium, and carrying out anaerobic culture at 37 ℃ for overnight to obtain a purified culture solution for 16S rDNA strain identification. Obtaining thalli by centrifuging the pure culture solution, extracting genome DNA, and carrying out PCR by adopting a bacterial 16S rDNA universal primer:
27F：AGAGTTTGATCCTGGCTCAG；
1492R：TACGGCTACCTTGTTACGACTT
sequencing the amplified product, comparing with NCBI database, and comparing the amplified product with NCBI database to obtain the result that the nucleic acid sequence similarity with bifidobacterium longum subspecies is up to 99.71%, and the amplified product is named as bifidobacterium subspecies longum WH 2270.
Example 2: safety evaluation of Bifidobacterium longum
1) Genome analysis and drug susceptibility test of bifidobacterium longum
Extracting genome DNA of bifidobacterium longum WHH2270 strain, and completing genome sequencing by utilizing a BGI platform and a Nanopore platform. Assembling the Nanopore data by using Flye, and performing self-error correction on the assembled genome by using Medaka software and the Nanopore platform sequencing data; and further correcting the sequence by using the multipolish and Polca software and combining with high-quality BGI platform sequencing data to obtain a genome completion map. The results showed that bifidobacterium longum WHH2270 has a genome size of 2.544mb and a gc ratio of 60%, wherein the genome contains 12 rRNA and 2208 genes. The setA which is verified by experiments with sequences in the VFDB database is an analysis data set, the sequence similarity is more than or equal to 80%, the coverage is more than or equal to 70% as a screening standard, and the result shows that no virulence gene is detected. The results of the drug susceptibility test showed that bifidobacterium longum WHH2270 was sensitive to ampicillin, vancomycin, gentamicin, streptomycin and chloramphenicol, and the results are shown in table 1.
TABLE 1 MIC values and drug sensitivity of bifidobacterium longum WH2270 against antibiotics
2) Animal tests for safety evaluation of bifidobacterium longum samples were administered to animals by both intraperitoneal injection and oral gavage, and the pathogenicity of the different exposure routes to the animals was evaluated by bifidobacterium longum WHH2270.
2.1 Intraperitoneal injection)
Inoculating activated Bifidobacterium longum WHH2270 with cysteine hydrochloride-containing MRS agar plate, anaerobic culturing, scraping colony on the plate, suspending in sterile physiological saline, mixing, adjusting bacterial suspension concentration with appropriate amount of sterile physiological saline, and making bacterial concentration in final bacterial suspension reach 5.0X10 7 CFU/mL for intraperitoneal injection in mice.
The number of mice is 40, and the male and female animals are half, and are randomly divided into 4 groups (each group is not less than 10), including a male mouse sterile physiological saline control group, a male mouse bacterial suspension group, a female mouse sterile physiological saline control group and a female mouse bacterial suspension group. Each mouse is injected with 0.2mL, namely the bacterial injection amount of each mouse in the test group is not less than 1.0X10 7 CFU. Animals were observed 1 time per day after intraperitoneal injection, at least for 21 days. The results are shown in Table 2.
TABLE 2 influence of bifidobacterium longum WHH2270 on mouse body weight and acute toxicity (intraperitoneal injection)
2.2 Oral lavage of stomach)
Bifidobacterium longum WHH2270 was inoculated into MRS broth containing cysteine hydrochloride and cultured anaerobically. Centrifuging to collect strain, re-suspending with sterile physiological saline to obtain final bacterial concentration of 2.5×10 8 CFU/mL bacterial suspension stock solution and bacterial final concentration reaching 1.25X10 9 CFU/mL concentrated bacterial suspension. One part of the supernatant after centrifugation is used as a supernatant stock solution, and the other part of the supernatant is frozen and concentrated to one fifth of the original volume, and the supernatant is concentrated as 5 times.
The number of mice is 80, the male and female halves are randomly divided into 8 groups (each group is not less than 10), and the groups comprise a male mouse culture medium control group, a male mouse bacterial suspension group, a male mouse 5-time concentrated culture medium control group and a male mouse 5-time concentrated bacterial suspension group; female mouse culture medium control group, female mouse bacterial suspension group, female mouse 5-fold concentrated culture medium control group and female mouse 5-fold concentrated bacterial suspension group. The mice in each group are filled with stomach in a volume of 20 mL/kg.BW, the stomach is continuously filled for 3d, the mice are fasted for overnight (16 h) before the first stomach filling, and the mice are fed for 3-4 h after the stomach filling. Animals were observed 1 time per day after oral gavage, for at least 21 days. The results are shown in Table 3.
TABLE 3 influence of bifidobacterium longum WHH2270 on body weight and acute toxicity (oral gavage)
Taken together, the results show that the injection of the stock solution of the bifidobacterium longum WHH2270 culture into the abdominal cavity has no influence on the body weight of the male mice and the female mice, and no toxic reaction or death of the mice occurs. The bifidobacterium longum WHH2270 culture stock solution and the concentrated solution were orally infused into the mice, and had no effect on the body weight of both male and female mice, and no toxic reaction or death of the mice was observed. The strain has high safety, no toxicity and no pathogenicity.
Example 3: animal test of Bifidobacterium longum subspecies WH 2270 for reducing blood sugar level by using tetraoxypyrimidine to induce insulin resistance glucose/lipid metabolism disorder rat model, and supplementing low dose of tetraoxypyrimidine (C 4 H 2 N 2 O 4 ·H 2 O, molecular weight 160.08), causes disorder of glucose/lipid metabolism, insulin resistance, and induces experimental diabetes. Detection of bifidobacterium longum subspecies longum WHH2270 pair sugarThe effects of blood sugar, blood lipid, and body weight in urine rats. The method comprises the following specific steps:
(1) Gastric lavage sample, feed preparation
(1.1) preparation of Bifidobacterium longum subspecies longum WH2270 powder
In a sterile environment, 1% of bifidobacterium longum subspecies longum WHH2270 glycerol frozen stock was inoculated in 10ml of liquid MRS medium and anaerobically cultured overnight at 37 ℃. The culture medium of the active generation was inoculated at 3% into 100ml of liquid MRS medium, and anaerobically cultured at 37℃overnight. The culture solution of the second generation is inoculated into 6L of liquid MRS culture medium of a fermentation tank at 3 percent, and is subjected to anaerobic static culture for 24 hours at 37 ℃. The culture solution was centrifuged at 7000rpm for 10min to collect the cells, and the cells were lyophilized after adding a protecting agent and mixing. The bacterial powder is collected and counted to be used as a sample for animal test gastric lavage. Preparing bacterial suspension, dissolving 2270 lyophilized powder in sterile water to obtain a solution with a concentration of 1.25X10 9 CFU/mL suspension, gavage volume 1mL.
(1.2) preparation of the Positive drug metformin
20mg of metformin is added into 1mL of sterile water to prepare a metformin preparation with the concentration of 20mg/mL, and the gastrolavage dosage is 200mg/kg, which is prepared for use at present.
(2) Feed stuff
(2.1) murine maintenance feed (ND): 40.6% of corn, 14% of wheat middling, 10% of wheat, 3% of alfalfa, 11% of soybean meal, 6% of fish meal, 6% of chicken meal, 4.4% of vitamins and light substances, 2% of bone protein powder, 1% of stone powder and 2% of salad oil.
(2.2) high heat energy feed (HFD): 10% of lard, 15% of sucrose, 15% of egg yolk powder, 5% of casein, 1.2% of cholesterol, 0.2% of sodium cholate, 0.6% of calcium bicarbonate, 0.4% of stone powder and 52.6% of mouse maintenance material.
(3) Animal experiment method (3.1) experimental animal: the invention adopts a tetraoxypyrimidine induced insulin resistance glucose/lipid metabolic disorder model, and selects SD male rats (150 g+/-20 g). The experimental rats were cycled for 10/14h in a barrier environment at 25+ -1deg.C and 50+ -5% relative humidity.
(3.2) grouping of animals: after 1 week of normal diet adaptation, basal blood glucose values were weighed and tested, randomly grouped. After 4 hours of fasting, tail blood was taken and fasting blood glucose was measured (0 hours). Blood glucose levels were measured at 0.5 and 2 hours after the administration of 2.5g/kg BW glucose. Statistical body weight and 0, 0.5, 2 hour blood glucose levels experimental rats were randomly divided into 5 groups of 12: blank Control (CK), MODEL control (MODEL), positive drug Metformin (MET) and 2 test groups (2270-ND and 2270-HFD).
(3.3) packet processing: CK group is normal diet + 1mL sterile physiological saline for gastric lavage; 2270-ND group is normal diet + 1mL of bacterial suspension for gastric lavage; MODEL group is high heat energy feed diet and 1mL sterile physiological saline for gastric lavage; MET group was a high heat feed diet + intragastric 1mL metformin solution; 2270-HFD group is a high heat energy feed plus 1mL of gastric lavage bacterial suspension. After grouping, 5 groups were given a normal diet for 1 week, and then the MRDOL group, MET group and 2270-HFD group were replaced with high heat feed. 3 weeks after HFD feeding, MRDOL, MET and 2270-HFD groups were fasted for 24 hours (without water), and were intraperitoneally injected with 105mg/kg BW of tetraoxapyrimidine at a rate of 1ml/100g body weight. The high heat feed was fed for 5 days after injection.
(4) And (3) index detection: at the end of the experiment, animals of each group were fasted for 4 hours and tested for fasting blood glucose, glucose tolerance, serum insulin, cholesterol and triglyceride levels.
(4.1) fasting blood glucose, glucose tolerance: after the test, each group of animals was fasted for 3-4 hours, and the fasting blood glucose (0 hour) blood glucose value was measured using a rogowski blood glucose meter and a matched test paper. The blood glucose of each group was measured by orally administering 2.5g/kg BW of glucose, and the blood glucose level of each group was measured at 0.5 and 2 hours after administration of glucose, and if the blood glucose level of the model control group was not less than 10mmol/L at 0.5 hour, or the blood glucose level was increased at any time point of 0.5 hour and 2 hours or the area under the blood glucose curve was increased in the model control group, the difference was significant as compared with the blank control group, and the model glucose metabolism disorder was judged to be established, and on this basis, the change in the fasting blood glucose, the blood glucose level after administration of glucose (0.5 and 2 hours) and the area under the blood glucose curve of 0, 0.5 and 2 hours was observed in the model control group and the test sample group.
Area under the blood glucose curve = [ (fasting blood glucose+0.5 hour blood glucose) ×0.5+ (2 hour blood glucose+0.5 hour blood glucose) ×1.5]/2.
(4.2) serum insulin: after the experiment, the rats were anesthetized, the heart was bled, and serum was collected by centrifugation. The insulin content in the serum was determined with reference to the kit instructions.
Insulin resistance index = serum insulin (uIU/mL) x fasting blood glucose (mmol/L)/22.5.
(4.3) cholesterol, triglycerides: after the experiment is finished, the rats are anesthetized, the hearts are bled, serum is collected by centrifugation, and serum cholesterol and triglyceride are detected. If the serum cholesterol or triglyceride of the model control group is obviously increased, compared with the blank control group, the difference is obvious, and the establishment of the lipid metabolism disorder of the model is judged, and on the basis, the blood lipid change of the model control group and the blood lipid change of the tested sample group are observed.
(4.4) determination of intestinal flora: after the test is finished, collecting the rat feces, extracting total DNA in the feces, amplifying by using a 16S rDNA v3+v4 region primer, sequencing on a machine, and collecting data for analysis.
(5) Experimental results
(5.1) modeling success index of insulin-resistant glycolipid disorder diabetic rat model: comparing the fasting blood glucose, blood lipid and serum insulin of rats in the model group and the blank control group, and calculating the area under the oral glucose tolerance curve and the insulin resistance index. Specific data are shown in table 4, the oral glucose of the model group is 0.5h and 2h, the area under the curve is extremely higher than that of the blank control group, the cholesterol level of the model group is extremely higher than that of the blank control group, the insulin resistance index is not different, and the success of modeling of the high-energy feed combined with the tetraoxypyrimidine-induced rat insulin resistance glucose/lipid metabolism disorder model is judged, so that experimental diabetes is induced.
TABLE 4 high energy feed combined with tetraoxypyrimidine induced glucose/lipid metabolism disorder model modeling success index for rats
| Model group | Blank control group | |
| Fasting blood glucose (mmol/L) | 6.38 | 6.39 |
| Glucose was orally administered for 0.5h blood glucose (mmol/L) | 11.35*** | 8.55 |
| Glucose 2h after oral administration (mmol/L) | 8.99*** | 6.61 |
| Area under curve (mmol/(L.times.h)) | 19.69*** | 15.11 |
| Serum TC (mM) | 9487.50*** | 1443.33 |
| Serum TG (mM) | 1346.36 | 1533.57 |
| Serum insulin (mIU/L) | 25.68 | 25.82 |
| Insulin resistance index | 7.28 | 7.33 |
And comparing the significance analysis of the model group with that of the blank control group. * P < 0.001
(5.2) bifidobacterium longum WHH2270 intervention affects body weight in diabetic rats with insulin-resistant glycolipid disorders: after grouping the 5 groups of rats, two of the CK and 2270-ND groups ingested normal maintenance diet, and three of the modem, 2270-HFD and MET groups ingested high energy diet. No significant differences in body weight were seen in the 5 groups of rats throughout the experimental period, as shown in fig. 1. Comparing the feeding of the five groups of rats, it was found that the feeding of the three groups of rats that ingest the high energy feed was significantly reduced compared to the feeding of the two groups of CK and 2270-ND, possibly because the high energy feed was too energetic or the taste was not favored by the rats. Despite the reduced intake of high energy feed, there was no significant difference in body weight from group to group throughout the experiment because of the higher energy value of the feed.
Blood sugar: the blood glucose levels of the five groups of rats were measured before the sample intervention in the rats, and as shown in fig. 2, there was no difference between the fasting blood glucose levels and the blood glucose levels of the rats of each group at 0.5 and 2 hours after glucose administration. After completion of gastric lavage for 8 weeks, fasting blood glucose and glucose tolerance test detection was performed. The results are shown in fig. 3, with no significant differences among the fasting blood glucose model group, the blank group and the experimental groups. After 0.5h of glucose oral intake, the blood glucose values of the bifidobacterium longum WHH2270 intervention group were significantly lower than those of the model group. After 2h of glucose oral administration, while there was no significant difference in blood glucose levels in WHH2270 intervention groups from the model group, there was a trend toward decrease. The area under the curve of blood glucose was calculated and the area under the curve of WHH2270 treatment group was significantly lower than that of the model group. The results show that the intervention of the bifidobacterium longum WH 2270 can obviously improve the glucose tolerance of diabetic rats and restore the regulating capacity of the rats on blood sugar. For normal diet rats, there was no difference in blood glucose from the blank group after WHH2270 intervention (2270-ND), indicating that bifidobacterium longum WHH2270 has sufficient safety in hypoglycemic function and does not cause hypoglycemia.
Blood lipid: serum cholesterol and triglycerides were detected 8 weeks after gastric lavage was completed. As a result, as shown in fig. 4, the high-energy feed-fed model group had significantly increased serum cholesterol levels and no significant difference in serum triglycerides compared to the blank group maintained feed feeding. There was no difference in WHH2270 dry prognosis at cholesterol level compared to the model group, and there was a trend but no significance in serum triglycerides at triglyceride level after WHH2270 dry prognosis.
Intestinal flora: the results of analysis compared the intestinal flora of rats in the placebo, model and WHH2270 intervention groups are shown in figure 5. In the alpha diversity analysis, the model group fed high energy diet and the WHH2270 intervention group, the alpha diversity was significantly lower than the blank control group fed maintenance diet, and there was no significant difference between the WHH2270 intervention group and the model group. Further in the comparison of phylum and genus levels, WHH2270 dry prognosis was found with significantly higher abundance of wart and actinomycetes phylum than model group, and significantly higher abundance of beneficial bifidobacteria and AKK than model group. The results show that bifidobacterium longum WHH2270 can effectively improve the intestinal flora structure of diabetic rats and increase beneficial intestinal bacteria, such as: abundance of bifidobacteria and AKK bacteria.
Short chain fatty acids: the short chain fatty acids, acetic acid and propionic acid, and acetic acid in serum were detected in each group of rat feces by gas phase method. The results are shown in FIG. 6. WHH2270 is dry, with significantly higher levels of acetic acid and propionic acid in the feces than in the model group, and significantly higher levels of acetic acid in the serum than in the model group. It was demonstrated that bifidobacterium longum WHH2270 can significantly increase the short chain fatty acid content in the feces and serum of diabetic rats.
G protein-coupled receptor: studies have shown that short chain fatty acids exert biological effects primarily through G protein-coupled receptors (GPR) 43 and 41. The combination of short chain fatty acids and GPRs stimulates secretion of the intestinal hormones GLP-1 and PYY, both of which act to lower blood glucose and thereby alleviate the occurrence of diabetes. The expression of the GPR41 and GPR43 genes in the intestinal tract was detected by fluorescent quantitative PCR, and as shown in fig. 7, the expression level of GPR43 gene in the intestinal tract of the model group of rats was found to be significantly lower than that of the control group, whereas the WHH2270 dry prognosis significantly increased GPR43 gene, which corresponds to the significantly increased expression of GPR41 and GPR43 genes in the pancreas of rats after WHH2270 dry prognosis. The results show that WH 2270 intervenes in diabetic rats, and can synchronously improve the expression level of the G protein coupled receptor gene of the mediated short chain fatty acid while improving the content of the short chain fatty acid, thereby achieving the effect of reducing blood sugar.
The raw materials and equipment used in the invention are common raw materials and equipment in the field unless specified otherwise; the methods used in the present invention are conventional in the art unless otherwise specified.
The foregoing description is only a preferred embodiment of the present invention, and is not intended to limit the present invention, and any simple modification, variation and equivalent transformation of the above embodiment according to the technical substance of the present invention still fall within the scope of the technical solution of the present invention.
Claims (10)
1. A bifidobacterium longum strain with blood glucose reducing effect, which is characterized in that: named WHH2270, and the microorganism classification named Bifidobacterium longum subspeciesBifidobacterium longum subsp. LongumThe method comprises the steps of carrying out a first treatment on the surface of the Has been deposited at the microorganism strain collection of Guangdong province at 10.18 of 2022, with the deposit address of Hirschner No. 100 in Guangzhou City and the deposit number of GDMCC No:62900.
2. use of bifidobacterium longum as claimed in claim 1 in the manufacture of a composition having the effect of improving the abundance of intestinal beneficial bacteria.
3. The use according to claim 2, wherein: the beneficial bacteria comprise bifidobacteria and AKK bacteria.
4. Use of bifidobacterium longum as claimed in claim 1 in the manufacture of a composition having the effect of increasing the short chain fatty acid content in the gut and serum.
5. The use according to claim 4, wherein: the short chain fatty acids are acetic acid and propionic acid.
6. Use of bifidobacterium longum as claimed in claim 1 in the manufacture of a composition having the effect of elevating the level of expression of a G protein-coupled receptor gene.
7. Use of bifidobacterium longum as claimed in claim 1 for the preparation of a composition having the effect of promoting secretion of the intestinal hormone GLP-1 and/or PYY.
8. Use of bifidobacterium longum as claimed in claim 1 in the preparation of a composition having hypoglycemic efficacy.
9. Use according to one of claims 2 to 8, characterized in that: the composition is powder, tablet, capsule, granule, pill or solution.
10. A probiotic bacterial agent, characterized in that: the bacterial liquid obtained after culturing bifidobacterium longum according to claim 1, or the cryogenic particles or dry powder bacterial agent obtained by further processing the bacterial liquid.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211588301.1A CN116376740A (en) | 2022-12-08 | 2022-12-08 | Bifidobacterium longum with blood sugar reducing effect and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202211588301.1A CN116376740A (en) | 2022-12-08 | 2022-12-08 | Bifidobacterium longum with blood sugar reducing effect and application thereof |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN116376740A true CN116376740A (en) | 2023-07-04 |
Family
ID=86971824
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202211588301.1A Pending CN116376740A (en) | 2022-12-08 | 2022-12-08 | Bifidobacterium longum with blood sugar reducing effect and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116376740A (en) |
Cited By (1)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116814501A (en) * | 2023-07-17 | 2023-09-29 | 微康益生菌(苏州)股份有限公司 | Bifidobacterium longum subspecies capable of relieving obesity and application thereof |
-
2022
- 2022-12-08 CN CN202211588301.1A patent/CN116376740A/en active Pending
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN116814501A (en) * | 2023-07-17 | 2023-09-29 | 微康益生菌(苏州)股份有限公司 | Bifidobacterium longum subspecies capable of relieving obesity and application thereof |
| CN116814501B (en) * | 2023-07-17 | 2024-02-09 | 微康益生菌(苏州)股份有限公司 | Bifidobacterium longum subspecies capable of relieving obesity and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| EP1885383B1 (en) | Feline probiotic bifidobacteria | |
| CN110964650B (en) | Bacterial strain for preventing and treating metabolic diseases and application thereof | |
| CN113293113B (en) | Bifidobacterium longum MI-186 and application thereof | |
| CN102274245B (en) | Novel lactacidophilus, composition thereof and use thereof in preparation of medicines for relieving diabetes mellitus and complications | |
| CN102935092B (en) | Novel lactobacilli and composition thereof, and application thereof in preparing medicines used for ameliorating diabetes and complications thereof | |
| CN110150669B (en) | Probiotic composition suitable for diabetic patients and application thereof | |
| JP5554994B2 (en) | Lactic acid bacteria-containing preparation | |
| JP2018532779A (en) | Felicaribacterium prausnitzi and desulfobibriopigel for use in the treatment or prevention of diabetes and bowel disease | |
| CN107106584A (en) | Synthetic composition for treating dysbolism | |
| WO2021098755A1 (en) | New application of bifidobacterium lactis bl-99 in inhibiting intestinal inflammation | |
| CN112458007A (en) | Lactobacillus crispatus for preventing and/or treating diseases related to genital tract flora disorder | |
| CN111996153A (en) | Bifidobacterium breve and application thereof | |
| CN113444668A (en) | Bacillus coagulans with hypoglycemic effect and application thereof | |
| TW201705969A (en) | Novel Lactobacillus mali APS1 and use thereof | |
| CN113308421A (en) | Lactobacillus plantarum BUFX and application thereof in metabolic syndrome | |
| CN112980725A (en) | Bifidobacterium lactis and application thereof in promoting growth and development of children and teenagers | |
| CN113797232B (en) | Composition with insulin resistance relieving function and application thereof | |
| CN116376740A (en) | Bifidobacterium longum with blood sugar reducing effect and application thereof | |
| CN106389478B (en) | Application of bacteroides fragilis in treatment and/or prevention of obesity or diabetes | |
| WO2017061353A1 (en) | Evaluation method, screening method, and production method for substances that inhibit rises in blood sugar values due to sucrose ingestion | |
| CN113164530A (en) | Application of enterococcus faecalis | |
| JP6301024B2 (en) | Felicaribacterium spp. | |
| KR101238836B1 (en) | Phamaceutical or food composition for treating or preventing obesity disease comprising a mixture of lactic acid bacteria | |
| CN115074276B (en) | Clostridium praecox capable of relieving non-alcoholic fatty liver disease and application thereof | |
| CN113005066B (en) | Compound bifidobacterium preparation for resisting allergy, increasing immunity, reducing blood sugar and fat and losing weight and preparation method thereof |
Legal Events
| Date | Code | Title | Description |
|---|---|---|---|
| PB01 | Publication | ||
| PB01 | Publication | ||
| SE01 | Entry into force of request for substantive examination | ||
| SE01 | Entry into force of request for substantive examination |