CN116286551B - Application of bifidobacterium longum subspecies infantis in regulating in-vivo fat metabolism, shaping, reducing fat and improving obesity - Google Patents
Application of bifidobacterium longum subspecies infantis in regulating in-vivo fat metabolism, shaping, reducing fat and improving obesity Download PDFInfo
- Publication number
- CN116286551B CN116286551B CN202310453211.XA CN202310453211A CN116286551B CN 116286551 B CN116286551 B CN 116286551B CN 202310453211 A CN202310453211 A CN 202310453211A CN 116286551 B CN116286551 B CN 116286551B
- Authority
- CN
- China
- Prior art keywords
- bifidobacterium longum
- nku
- mice
- longum subspecies
- regulating
- 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.)
- Active
Links
- 241001608472 Bifidobacterium longum Species 0.000 title claims abstract description 127
- 229940009291 bifidobacterium longum Drugs 0.000 title claims abstract description 127
- 208000008589 Obesity Diseases 0.000 title claims abstract description 53
- 235000020824 obesity Nutrition 0.000 title claims abstract description 53
- 230000001105 regulatory effect Effects 0.000 title claims abstract description 35
- 230000004060 metabolic process Effects 0.000 title claims abstract description 34
- 238000001727 in vivo Methods 0.000 title abstract description 30
- 238000007493 shaping process Methods 0.000 title description 2
- 239000003814 drug Substances 0.000 claims abstract description 19
- 229940079593 drug Drugs 0.000 claims abstract description 12
- 238000004321 preservation Methods 0.000 claims abstract description 10
- 238000009629 microbiological culture Methods 0.000 claims abstract description 5
- 241000699670 Mus sp. Species 0.000 claims description 116
- 230000000968 intestinal effect Effects 0.000 claims description 44
- 210000004369 blood Anatomy 0.000 claims description 38
- 239000008280 blood Substances 0.000 claims description 38
- 230000001580 bacterial effect Effects 0.000 claims description 29
- 201000010063 epididymitis Diseases 0.000 claims description 25
- 210000002966 serum Anatomy 0.000 claims description 23
- 150000002632 lipids Chemical class 0.000 claims description 22
- 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 claims description 18
- 244000005700 microbiome Species 0.000 claims description 18
- 241000186000 Bifidobacterium Species 0.000 claims description 17
- 235000009200 high fat diet Nutrition 0.000 claims description 16
- 241000699666 Mus <mouse, genus> Species 0.000 claims description 15
- 230000000694 effects Effects 0.000 claims description 15
- 230000012010 growth Effects 0.000 claims description 15
- 102000004889 Interleukin-6 Human genes 0.000 claims description 14
- 229940100601 interleukin-6 Drugs 0.000 claims description 14
- 210000004185 liver Anatomy 0.000 claims description 14
- 108060008682 Tumor Necrosis Factor Proteins 0.000 claims description 12
- 102000000852 Tumor Necrosis Factor-alpha Human genes 0.000 claims description 12
- 230000002757 inflammatory effect Effects 0.000 claims description 12
- 210000001789 adipocyte Anatomy 0.000 claims description 11
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 10
- 239000008103 glucose Substances 0.000 claims description 10
- 230000001737 promoting effect Effects 0.000 claims description 10
- 102000003777 Interleukin-1 beta Human genes 0.000 claims description 9
- 108090000193 Interleukin-1 beta Proteins 0.000 claims description 9
- 235000012000 cholesterol Nutrition 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 9
- 241000605909 Fusobacterium Species 0.000 claims description 8
- 108010023302 HDL Cholesterol Proteins 0.000 claims description 8
- 108010028554 LDL Cholesterol Proteins 0.000 claims description 8
- 241000192125 Firmicutes Species 0.000 claims description 7
- 230000006378 damage Effects 0.000 claims description 7
- LOGFVTREOLYCPF-KXNHARMFSA-N (2s,3r)-2-[[(2r)-1-[(2s)-2,6-diaminohexanoyl]pyrrolidine-2-carbonyl]amino]-3-hydroxybutanoic acid Chemical compound C[C@@H](O)[C@@H](C(O)=O)NC(=O)[C@H]1CCCN1C(=O)[C@@H](N)CCCCN LOGFVTREOLYCPF-KXNHARMFSA-N 0.000 claims description 6
- 241000589291 Acinetobacter Species 0.000 claims description 6
- 230000002401 inhibitory effect Effects 0.000 claims description 6
- 241000589989 Helicobacter Species 0.000 claims description 5
- 210000003934 vacuole Anatomy 0.000 claims description 5
- 238000012423 maintenance Methods 0.000 claims description 4
- 230000007863 steatosis Effects 0.000 claims description 4
- 231100000240 steatosis hepatitis Toxicity 0.000 claims description 4
- 150000003626 triacylglycerols Chemical class 0.000 claims description 4
- 210000003494 hepatocyte Anatomy 0.000 claims description 2
- 235000013402 health food Nutrition 0.000 claims 7
- 230000002906 microbiologic effect Effects 0.000 claims 3
- 208000030159 metabolic disease Diseases 0.000 claims 2
- 239000000825 pharmaceutical preparation Substances 0.000 claims 2
- 229940127557 pharmaceutical product Drugs 0.000 claims 2
- 241000605059 Bacteroidetes Species 0.000 claims 1
- 206010019708 Hepatic steatosis Diseases 0.000 claims 1
- 239000008186 active pharmaceutical agent Substances 0.000 claims 1
- 229940088679 drug related substance Drugs 0.000 claims 1
- 108010088383 interleukin-6 receptor alpha Proteins 0.000 claims 1
- 235000013376 functional food Nutrition 0.000 abstract description 5
- 230000000813 microbial effect Effects 0.000 abstract description 5
- 108090001005 Interleukin-6 Proteins 0.000 description 13
- 238000004458 analytical method Methods 0.000 description 13
- 239000003833 bile salt Substances 0.000 description 13
- 210000001035 gastrointestinal tract Anatomy 0.000 description 12
- 239000001963 growth medium Substances 0.000 description 12
- 238000012360 testing method Methods 0.000 description 12
- 239000007788 liquid Substances 0.000 description 11
- 238000013116 obese mouse model Methods 0.000 description 11
- 241000894006 Bacteria Species 0.000 description 9
- 241000588769 Proteus <enterobacteria> Species 0.000 description 9
- 210000005228 liver tissue Anatomy 0.000 description 9
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 8
- 239000000243 solution Substances 0.000 description 8
- 239000006228 supernatant Substances 0.000 description 8
- 241000606125 Bacteroides Species 0.000 description 7
- 108090000913 Nitrate Reductases Proteins 0.000 description 7
- 241000566145 Otus Species 0.000 description 7
- 230000008859 change Effects 0.000 description 7
- 238000000034 method Methods 0.000 description 7
- 230000004083 survival effect Effects 0.000 description 7
- 108020004414 DNA Proteins 0.000 description 6
- 206010018910 Haemolysis Diseases 0.000 description 6
- 239000002253 acid Substances 0.000 description 6
- 230000009286 beneficial effect Effects 0.000 description 6
- 230000037396 body weight Effects 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 230000002949 hemolytic effect Effects 0.000 description 6
- 230000002829 reductive effect Effects 0.000 description 6
- 208000024891 symptom Diseases 0.000 description 6
- 108020004465 16S ribosomal RNA Proteins 0.000 description 5
- 241000221955 Chaetomium Species 0.000 description 5
- 230000005526 G1 to G0 transition Effects 0.000 description 5
- 241000282414 Homo sapiens Species 0.000 description 5
- 230000003078 antioxidant effect Effects 0.000 description 5
- 239000011248 coating agent Substances 0.000 description 5
- 238000000576 coating method Methods 0.000 description 5
- 239000007787 solid Substances 0.000 description 5
- 241000894007 species Species 0.000 description 5
- 238000010186 staining Methods 0.000 description 5
- 239000000725 suspension Substances 0.000 description 5
- 241000192700 Cyanobacteria Species 0.000 description 4
- 206010061218 Inflammation Diseases 0.000 description 4
- CTQNGGLPUBDAKN-UHFFFAOYSA-N O-Xylene Chemical group CC1=CC=CC=C1C CTQNGGLPUBDAKN-UHFFFAOYSA-N 0.000 description 4
- 238000002835 absorbance Methods 0.000 description 4
- 210000000577 adipose tissue Anatomy 0.000 description 4
- 210000004027 cell Anatomy 0.000 description 4
- 238000010790 dilution Methods 0.000 description 4
- 239000012895 dilution Substances 0.000 description 4
- 239000002158 endotoxin Substances 0.000 description 4
- 230000008588 hemolysis Effects 0.000 description 4
- 230000004054 inflammatory process Effects 0.000 description 4
- 238000009630 liquid culture Methods 0.000 description 4
- 239000000203 mixture Substances 0.000 description 4
- 239000012188 paraffin wax Substances 0.000 description 4
- 238000012216 screening Methods 0.000 description 4
- 210000001519 tissue Anatomy 0.000 description 4
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 description 4
- 241000186361 Actinobacteria <class> Species 0.000 description 3
- VLSOAXRVHARBEQ-UHFFFAOYSA-N [4-fluoro-2-(hydroxymethyl)phenyl]methanol Chemical compound OCC1=CC=C(F)C=C1CO VLSOAXRVHARBEQ-UHFFFAOYSA-N 0.000 description 3
- 230000033228 biological regulation Effects 0.000 description 3
- 238000004113 cell culture Methods 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 238000011161 development Methods 0.000 description 3
- 238000002474 experimental method Methods 0.000 description 3
- 230000002496 gastric effect Effects 0.000 description 3
- 244000005709 gut microbiome Species 0.000 description 3
- 239000002054 inoculum Substances 0.000 description 3
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 3
- 230000002503 metabolic effect Effects 0.000 description 3
- NLKNQRATVPKPDG-UHFFFAOYSA-M potassium iodide Chemical compound [K+].[I-] NLKNQRATVPKPDG-UHFFFAOYSA-M 0.000 description 3
- 238000000513 principal component analysis Methods 0.000 description 3
- 239000006041 probiotic Substances 0.000 description 3
- 235000018291 probiotics Nutrition 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 238000005406 washing Methods 0.000 description 3
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 3
- 230000004584 weight gain Effects 0.000 description 3
- 235000019786 weight gain Nutrition 0.000 description 3
- 239000008096 xylene Substances 0.000 description 3
- MINDHVHHQZYEEK-UHFFFAOYSA-N (E)-(2S,3R,4R,5S)-5-[(2S,3S,4S,5S)-2,3-epoxy-5-hydroxy-4-methylhexyl]tetrahydro-3,4-dihydroxy-(beta)-methyl-2H-pyran-2-crotonic acid ester with 9-hydroxynonanoic acid Natural products CC(O)C(C)C1OC1CC1C(O)C(O)C(CC(C)=CC(=O)OCCCCCCCCC(O)=O)OC1 MINDHVHHQZYEEK-UHFFFAOYSA-N 0.000 description 2
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 241000186015 Bifidobacterium longum subsp. infantis Species 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- -1 DPPH free radical Chemical class 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 2
- 241001112693 Lachnospiraceae Species 0.000 description 2
- 208000001145 Metabolic Syndrome Diseases 0.000 description 2
- 229910002651 NO3 Inorganic materials 0.000 description 2
- NHNBFGGVMKEFGY-UHFFFAOYSA-N Nitrate Chemical compound [O-][N+]([O-])=O NHNBFGGVMKEFGY-UHFFFAOYSA-N 0.000 description 2
- 241001547082 Pseudobacillus Species 0.000 description 2
- 201000000690 abdominal obesity-metabolic syndrome Diseases 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 238000003556 assay Methods 0.000 description 2
- 229940093761 bile salts Drugs 0.000 description 2
- 239000007382 columbia agar Substances 0.000 description 2
- 238000001514 detection method Methods 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- MGJZITXUQXWAKY-UHFFFAOYSA-N diphenyl-(2,4,6-trinitrophenyl)iminoazanium Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1N=[N+](C=1C=CC=CC=1)C1=CC=CC=C1 MGJZITXUQXWAKY-UHFFFAOYSA-N 0.000 description 2
- LOKCTEFSRHRXRJ-UHFFFAOYSA-I dipotassium trisodium dihydrogen phosphate hydrogen phosphate dichloride Chemical compound P(=O)(O)(O)[O-].[K+].P(=O)(O)([O-])[O-].[Na+].[Na+].[Cl-].[K+].[Cl-].[Na+] LOKCTEFSRHRXRJ-UHFFFAOYSA-I 0.000 description 2
- 201000010099 disease Diseases 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- HHEAADYXPMHMCT-UHFFFAOYSA-N dpph Chemical compound [O-][N+](=O)C1=CC([N+](=O)[O-])=CC([N+]([O-])=O)=C1[N]N(C=1C=CC=CC=1)C1=CC=CC=C1 HHEAADYXPMHMCT-UHFFFAOYSA-N 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 229920006008 lipopolysaccharide Polymers 0.000 description 2
- 210000005229 liver cell Anatomy 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 239000011259 mixed solution Substances 0.000 description 2
- 238000010172 mouse model Methods 0.000 description 2
- 229960003128 mupirocin Drugs 0.000 description 2
- 229930187697 mupirocin Natural products 0.000 description 2
- DDHVILIIHBIMQU-YJGQQKNPSA-L mupirocin calcium hydrate Chemical compound O.O.[Ca+2].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.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 DDHVILIIHBIMQU-YJGQQKNPSA-L 0.000 description 2
- 235000016709 nutrition Nutrition 0.000 description 2
- 239000002953 phosphate buffered saline Substances 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 108090000623 proteins and genes Proteins 0.000 description 2
- 238000000746 purification Methods 0.000 description 2
- 238000011160 research Methods 0.000 description 2
- 238000011144 upstream manufacturing Methods 0.000 description 2
- BHQCQFFYRZLCQQ-UHFFFAOYSA-N (3alpha,5alpha,7alpha,12alpha)-3,7,12-trihydroxy-cholan-24-oic acid Natural products OC1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 BHQCQFFYRZLCQQ-UHFFFAOYSA-N 0.000 description 1
- 241000193830 Bacillus <bacterium> Species 0.000 description 1
- 108020000946 Bacterial DNA Proteins 0.000 description 1
- 241000186018 Bifidobacterium adolescentis Species 0.000 description 1
- 241001134770 Bifidobacterium animalis Species 0.000 description 1
- 241000186016 Bifidobacterium bifidum Species 0.000 description 1
- 241000186012 Bifidobacterium breve Species 0.000 description 1
- 108090000994 Catalytic RNA Proteins 0.000 description 1
- 102000053642 Catalytic RNA Human genes 0.000 description 1
- 239000004380 Cholic acid Substances 0.000 description 1
- 102000016928 DNA-directed DNA polymerase Human genes 0.000 description 1
- 108010014303 DNA-directed DNA polymerase Proteins 0.000 description 1
- 208000036649 Dysbacteriosis Diseases 0.000 description 1
- 208000027244 Dysbiosis Diseases 0.000 description 1
- 208000032928 Dyslipidaemia Diseases 0.000 description 1
- 238000002965 ELISA Methods 0.000 description 1
- 108010000912 Egg Proteins Proteins 0.000 description 1
- 102000002322 Egg Proteins Human genes 0.000 description 1
- 235000019733 Fish meal Nutrition 0.000 description 1
- 241001453172 Fusobacteria Species 0.000 description 1
- 241000287828 Gallus gallus Species 0.000 description 1
- 108010002352 Interleukin-1 Proteins 0.000 description 1
- 102000000589 Interleukin-1 Human genes 0.000 description 1
- 208000017170 Lipid metabolism disease Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 108091028043 Nucleic acid sequence Proteins 0.000 description 1
- 239000005662 Paraffin oil Substances 0.000 description 1
- 229930040373 Paraformaldehyde Natural products 0.000 description 1
- QGMRQYFBGABWDR-UHFFFAOYSA-M Pentobarbital sodium Chemical compound [Na+].CCCC(C)C1(CC)C(=O)NC(=O)[N-]C1=O QGMRQYFBGABWDR-UHFFFAOYSA-M 0.000 description 1
- 241000192142 Proteobacteria Species 0.000 description 1
- 230000002292 Radical scavenging effect Effects 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 241000191967 Staphylococcus aureus Species 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 241000209140 Triticum Species 0.000 description 1
- 235000021307 Triticum 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
- 230000002159 abnormal effect Effects 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000002411 adverse Effects 0.000 description 1
- 229940024606 amino acid Drugs 0.000 description 1
- 235000001014 amino acid Nutrition 0.000 description 1
- 150000001413 amino acids Chemical class 0.000 description 1
- 230000003444 anaesthetic effect Effects 0.000 description 1
- 238000010171 animal model Methods 0.000 description 1
- 229940118852 bifidobacterium animalis Drugs 0.000 description 1
- 229940002008 bifidobacterium bifidum Drugs 0.000 description 1
- 229940004120 bifidobacterium infantis Drugs 0.000 description 1
- 238000007622 bioinformatic analysis Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 239000006161 blood agar Substances 0.000 description 1
- 238000004422 calculation algorithm Methods 0.000 description 1
- 210000004534 cecum Anatomy 0.000 description 1
- 239000006285 cell suspension Substances 0.000 description 1
- 230000001413 cellular effect Effects 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- BHQCQFFYRZLCQQ-OELDTZBJSA-N cholic acid Chemical compound 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 BHQCQFFYRZLCQQ-OELDTZBJSA-N 0.000 description 1
- 229960002471 cholic acid Drugs 0.000 description 1
- 235000019416 cholic acid Nutrition 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 230000001276 controlling effect Effects 0.000 description 1
- 235000005822 corn Nutrition 0.000 description 1
- 230000002596 correlated effect Effects 0.000 description 1
- 230000001351 cycling effect Effects 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 230000018044 dehydration Effects 0.000 description 1
- 238000006297 dehydration reaction Methods 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 238000004925 denaturation Methods 0.000 description 1
- 230000036425 denaturation Effects 0.000 description 1
- KXGVEGMKQFWNSR-UHFFFAOYSA-N deoxycholic acid Natural products C1CC2CC(O)CCC2(C)C2C1C1CCC(C(CCC(O)=O)C)C1(C)C(O)C2 KXGVEGMKQFWNSR-UHFFFAOYSA-N 0.000 description 1
- 230000001627 detrimental effect Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 230000035622 drinking Effects 0.000 description 1
- 230000007140 dysbiosis Effects 0.000 description 1
- 235000013345 egg yolk Nutrition 0.000 description 1
- 210000002969 egg yolk Anatomy 0.000 description 1
- 230000037149 energy metabolism Effects 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 238000001914 filtration Methods 0.000 description 1
- 239000004467 fishmeal Substances 0.000 description 1
- 238000009472 formulation Methods 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 238000012268 genome sequencing Methods 0.000 description 1
- 230000005484 gravity Effects 0.000 description 1
- 230000008821 health effect Effects 0.000 description 1
- 238000007489 histopathology method Methods 0.000 description 1
- 230000015784 hyperosmotic salinity response Effects 0.000 description 1
- 210000003405 ileum Anatomy 0.000 description 1
- 230000036039 immunity Effects 0.000 description 1
- 238000011065 in-situ storage Methods 0.000 description 1
- 238000011534 incubation Methods 0.000 description 1
- 230000001939 inductive effect Effects 0.000 description 1
- 230000008595 infiltration Effects 0.000 description 1
- 238000001764 infiltration Methods 0.000 description 1
- 229910052500 inorganic mineral Inorganic materials 0.000 description 1
- 230000007413 intestinal health Effects 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 230000003902 lesion Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 230000037356 lipid metabolism Effects 0.000 description 1
- 239000002075 main ingredient Substances 0.000 description 1
- 235000012054 meals Nutrition 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- 239000002207 metabolite Substances 0.000 description 1
- 239000011707 mineral Substances 0.000 description 1
- 235000010755 mineral Nutrition 0.000 description 1
- 238000002156 mixing Methods 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
- 239000006872 mrs medium Substances 0.000 description 1
- 239000013642 negative control Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 238000004806 packaging method and process Methods 0.000 description 1
- 229920002866 paraformaldehyde Polymers 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 230000007170 pathology Effects 0.000 description 1
- 229960002275 pentobarbital sodium Drugs 0.000 description 1
- 239000000902 placebo Substances 0.000 description 1
- 229940068196 placebo Drugs 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 230000005180 public health Effects 0.000 description 1
- 230000000306 recurrent effect Effects 0.000 description 1
- 210000005000 reproductive tract Anatomy 0.000 description 1
- 108091092562 ribozyme Proteins 0.000 description 1
- 238000007789 sealing Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 235000012424 soybean oil Nutrition 0.000 description 1
- 239000003549 soybean oil Substances 0.000 description 1
- 230000007480 spreading Effects 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 230000001954 sterilising effect Effects 0.000 description 1
- 238000004659 sterilization and disinfection Methods 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000002459 sustained effect Effects 0.000 description 1
- 230000002195 synergetic effect 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
Classifications
-
- 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
- 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
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/04—Anorexiants; Antiobesity agents
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P3/00—Drugs for disorders of the metabolism
- A61P3/06—Antihyperlipidemics
-
- 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
-
- 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
-
- 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 discloses an application of bifidobacterium longum subspecies infantis NKU FB3-14 in preparing functional foods or medicines for obesity and/or regulating in-vivo fat metabolismBifidobacterium longum subsp.infantis) The microbial strain is preserved in China general microbiological culture Collection center, north Chen Xili No. 1, 3 of the Korean area of Beijing city, and the preservation number is: CGMCC No.25762, the preservation date is 2022, 9 and 21. The bifidobacterium longum subspecies infantis NKU FB3-14 can be used for preparing functional foods or medicines for obesity and/or regulating in-vivo fat metabolism.
Description
Technical Field
The invention relates to the technical field of microorganisms, in particular to application of bifidobacterium longum subspecies infancy NKU FB3-14 in relieving obesity and regulating in-vivo fat metabolism.
Background
The world obesity association recognizes obesity as a chronic recurrent disease, one of the major problems facing the world public health sector today, and the number of obese people has exceeded 6.5 billion. The growing trend for obese people is more pronounced in developing countries. Many diseases caused by obesity can have adverse health effects and increase mortality. The intestinal micro-ecosystem has a regulating effect on lipid metabolism, and how to change obesity by adjusting the intestinal micro-ecosystem has become a concern for human beings. The probiotics can regulate the flora composition in the intestinal canal, make the beneficial bacteria become dominant flora, inhibit the growth of harmful bacteria and the generation of related metabolites, participate in the immune process of the organism, and can effectively relieve obesity symptoms. Thus, intervention of intestinal micro-ecosystem by probiotics and prevention of obesity have become one of the hot spots of research in the modern nutritional and health field.
Bifidobacteria are widely distributed in the digestive tract, oral cavity and genital tract of human body and are one of the main genera of intestinal microorganisms. Among them, some bifidobacteria have been used as probiotics, such as bifidobacterium adolescentis, bifidobacterium animalis, bifidobacterium bifidum, bifidobacterium breve, bifidobacterium infantis, bifidobacterium longum, etc., which play an important role in intestinal health, energy metabolism and immunity. The bifidobacteria are changed in the human intestinal canal at any time, and the bifidobacteria in the human intestinal canal have larger differences in different age groups. However, during the whole life cycle, bifidobacteria in the intestinal tract maintain dynamic balance, which is important for the normal function of the intestinal tract and the health of human bodies. Therefore, it is necessary to screen out bifidobacteria which are capable of regulating intestinal flora and effectively relieving obesity.
Disclosure of Invention
The invention is realized by the following technical scheme:
the invention provides an application of bifidobacterium longum subspecies infancy NKU FB3-14 in preparing functional foods or medicines for obesity and/or regulating in-vivo fat metabolism, wherein the bifidobacterium longum subspecies infancy is @ or @ isBifidobacterium longum subsp. infantis) The microbial strain is preserved in China general microbiological culture Collection center, north Chen Xili No. 1, 3 of the Korean area of Beijing city, and the preservation number is: CGMCC No.25762, the preservation date is 2022, 9 and 21.
The bifidobacterium longum subspecies infantis NKU FB3-14 provided by the invention has excellent acid resistance, bile salt resistance and oxidation resistance, and the strain is proved to be a safe strain with excellent functional characteristics of relieving obesity and regulating fat metabolism in vivo by multi-aspect index measurement.
Specifically, the form of the thallus displayed by the Bifidobacterium longum subspecies infantis NKU FB3-14 scanning electron microscope comprises one or more of V-type, Y-type and ball rod type.
Specifically, the bifidobacterium longum subspecies infantis NKU FB3-14 is a gram positive bacterium.
Specifically, the colony of the bifidobacterium longum subspecies infantis NKU FB3-14 is characterized in that the colony is tiny and smooth, is milky white, has neat and raised edges, and has soft texture.
Specifically, the growth characteristic of the bifidobacterium longum subspecies infantis NKU FB3-14 is that the bifidobacterium subspecies infantis is grown anaerobically at the temperature of 35-37 ℃ and enters a stable period after being cultured for 24-36 hours.
Specifically, the bifidobacterium longum subspecies infantis NKU FB3-14 does not contain an active nitrate reductase. The strain did not undergo a color reaction in the nitrate reductase activity test.
In particular, the bifidobacterium longum subspecies infantis NKU FB3-14 is gamma hemolysis without hemolytic activity. Growth in Columbia agar plates was free of hemolysis and the results confirm that the bacteria were safe strains.
In particular, the bifidobacterium longum subspecies infantis NKU FB3-14 has acid resistance, and particularly has the survival rate of not less than 145% in a culture environment with the pH of 4+/-0.3.
Specifically, the bifidobacterium longum subspecies infantis NKU FB3-14 has anti-bile salt characteristics, and particularly the survival rate of the bifidobacterium longum subspecies infantis NKU FB3-14 is not less than 150% in a culture environment with the mass ratio of bile salt of 0.05+/-0.01%.
In particular, the cell culture suspension, cell-free extract and cell-free supernatant of the bifidobacterium longum subspecies infantis NKU FB3-14 have antioxidant properties, and in particular, the cell-free supernatant has a DPPH free radical clearance of not less than 90%.
In particular, the bifidobacterium longum subspecies infantis NKU FB3-14 is used for preparing obesity and/or regulating fat metabolism in vivo by inhibiting the weight proportion of liver and/or epididymal fat mass to body weight so as to relieve obesity and/or regulate fat metabolism in vivo.
In particular, the bifidobacterium longum subspecies infancy NKU FB3-14 is useful in the preparation of obesity and/or regulating fat metabolism in vivo by modulating blood glucose to relieve obesity and/or regulate fat metabolism in vivo.
In particular, the bifidobacterium longum subspecies infancy NKU FB3-14 is used for relieving obesity and/or regulating fat metabolism in vivo by regulating blood lipid metabolic disturbance in the preparation of obesity and/or regulating fat metabolism in vivo.
In particular, the application of bifidobacterium longum subspecies infancy NKU FB3-14 in preparing obesity and/or regulating in vivo fat metabolism relieves obesity and/or regulates in vivo fat metabolism by inhibiting the content of total cholesterol TC, total triglyceride TG and low-density lipoprotein cholesterol LDL-C in blood fat and promoting the increase of the content of high-density lipoprotein cholesterol HDL-C in blood fat.
In particular, the bifidobacterium longum subspecies infantis NKU FB3-14 is used for relieving obesity and/or regulating fat metabolism in vivo by regulating inflammatory factors in serum in the preparation of obesity and/or regulating fat metabolism in vivo.
In particular, the application of bifidobacterium longum subspecies infancy NKU FB3-14 in preparing obesity and/or regulating in vivo fat metabolism can relieve obesity and/or regulate in vivo fat metabolism by reducing the content of interleukin-1 beta (IL-1 beta), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in serum.
In particular, the bifidobacterium longum subspecies infancy NKU FB3-14 is useful in the preparation of obesity and/or modulating fat metabolism in vivo by alleviating high fat diet-induced histological damage.
In particular, the bifidobacterium longum subspecies infantis NKU FB3-14 is useful in the preparation of high fat diet-induced histological lesions by reducing expansion of adipocytes and reducing the number of adipocyte vacuoles.
In particular, the bifidobacterium longum subspecies infancy NKU FB3-14 is used for preparing obesity and/or regulating in-vivo fat metabolism by regulating the structure of a high-fat metabolism intestinal flora to relieve obesity and/or regulate in-vivo fat metabolism.
Specifically, the regulation of the structure of the hypersecretion intestinal flora is achieved by regulating the alpha diversity and beta diversity of the intestinal flora of mice.
Specifically, the Bifidobacterium longum subspecies NKU FB3-14 can inhibit the growth of the bacterial flora of the phylum Proteus and Fusobacterium at the phylum level by regulating the structure of the intestinal flora of mice in regulating the structure of the intestinal flora of the metabolism of high fat, and can relieve obesity by promoting the growth of the bacterial flora of the phylum Thick-wall phylum (Firmics), the phylum Bacteroides (Bacteroides) and the phylum Desulfobacilli (Desulfobacilli).
Specifically, the bifidobacterium longum subspecies infancy NKU FB3-14 promotes the chaetomium species to achieve the effect of promoting the chaetomium species at the genus level by adjusting the intestinal flora structure of mice in the process of adjusting the intestinal flora structure of high-fat metabolism Lachnospiraceae spp.), the group NK4A136 of the genus Muril, muril @Muribaculaceaespp..Bifidobacterium genus ]Bifidibacteriumspp. and helicobacter speciesOscillospiraceaespp.) and inhibit Acinetobacter genusAcinetobacterspp.).
The invention has the characteristics and beneficial effects that:
(1) In the model of obese mice, the bifidobacterium longum subspecies NKU FB3-14 can significantly relieve obesity and/or regulate fat metabolism in the body of the mice, and the evaluation indexes comprise determination of weight proportion of liver and/or epididymal fat mass of the mice, blood sugar, blood fat level, inflammatory factors and determination of change of intestinal flora structure of the mice; the use of Bifidobacterium longum subspecies infantis NKU FB3-14 can significantly reduce the weight specific gravity of the liver and/or epididymal fat mass of mice; the use of Bifidobacterium longum subspecies infantis NKU FB3-14 can significantly reduce blood glucose in mice; the use of Bifidobacterium longum subspecies NKU FB3-14 can regulate and control the blood lipid level of mice, inhibit the content of total cholesterol TC, total triglyceride TG and low-density lipoprotein cholesterol LDL-C in blood lipid and promote the content of high-density lipoprotein cholesterol HDL-C in blood lipid; the use of Bifidobacterium longum subspecies infantis NKU FB3-14 can regulate and control inflammatory factors in serum of mice, and can reduce the contents of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in the serum; the use of Bifidobacterium longum subspecies infantis NKU FB3-14 can alleviate the histological damage caused by high fat diet by reducing the expansion of adipocytes and reducing the number of adipocyte vacuoles; the use of Bifidobacterium longum subspecies infantis NKU FB3-14 can promote the growth of the flora of the phylum Thick-walled bacteria (Firmics), the phylum Bacteroides (Bactoides), the phylum desulphus (Desulfobacillosis), inhibit the growth of the flora of the phylum Proteus and the phylum Fusobacterium, balance the flora structure in the intestinal tract of mice, increase the alpha diversity of the intestinal flora, normalize the intestinal flora and reduce the occurrence of obesity.
(2) The bifidobacterium longum subspecies infantis NKU FB3-14 can be used for preparing functional foods or medicines for obesity and/or regulating in-vivo fat metabolism.
Drawings
In order to more clearly illustrate the technical solutions of the embodiments of the present invention, the drawings that are needed in the embodiments or the description of the prior art will be briefly introduced below, and it is obvious that the drawings in the following description are only some embodiments of the present invention, and other drawings can be obtained according to these drawings without inventive effort for a person skilled in the art.
FIG. 1 is a diagram showing the morphology of Bifidobacterium longum subspecies infantis NKU FB3-14 according to the embodiments of the present invention.
FIG. 2 shows the construction of a Bifidobacterium longum subspecies infancy NKU FB3-14 phylogenetic tree according to the embodiments of the present invention.
FIG. 3 shows the experimental results of the activity of the nitrate reductase of Bifidobacterium longum subspecies infantis NKU FB3-14 provided by the embodiment of the invention.
FIG. 4 shows the results of a hemolysis test of Bifidobacterium longum subspecies infantis NKU FB3-14 according to the examples of the present invention.
FIG. 5 shows the results of acid resistance of Bifidobacterium longum subspecies infantis NKU FB3-14, wherein a in FIG. 5 is a culture environment with pH 4, and b in FIG. 5 is a culture environment with pH 3.
FIG. 6 shows the results of the anti-bile salt properties of Bifidobacterium longum subspecies infantis NKU FB3-14 according to the present invention, wherein a in FIG. 6 is a culture environment with a bile salt mass ratio of 0.05%, and b in FIG. 6 is a culture environment with a bile salt mass ratio of 0.1%.
FIG. 7 shows the antioxidant capacity of Bifidobacterium longum subspecies infantis NKU FB3-14 according to the examples of the present invention.
FIG. 8 is a graph showing the weight results of mice according to the embodiment of the present invention, wherein a in FIG. 8 is the weight change of mice of different groups over time; in fig. 8 b is the final body weight of the mice of the different groups.
Fig. 9 is a mass ratio of the liver weight and the epididymal weight of the mice provided by the embodiment of the invention, wherein a in fig. 9 is the mass ratio of the liver weight of the mice of different groups to the epididymal weight of the mice of different groups, and b in fig. 9 is the mass ratio of the epididymal weight of the mice of different groups to the epididymal weight of the mice of different groups.
Fig. 10 shows fasting blood glucose values of mice of different groups according to the present invention.
FIG. 11 is a graph showing the blood lipid level related index in serum of mice of different groups according to the present invention, wherein a in FIG. 11 is the total cholesterol TC content of the mice of different groups, b in FIG. 11 is the total triglyceride TG content of the mice of different groups, C in FIG. 11 is the low-density lipoprotein cholesterol LDL-C content of the mice of different groups, and d in FIG. 11 is the high-density lipoprotein cholesterol HDL-C content of the mice of different groups.
FIG. 12 shows the index of inflammatory factors and endotoxins in serum of mice of different groups according to the present invention, wherein a in FIG. 12 is the interleukin-1β (IL-1β) content of mice of different groups, b in FIG. 12 is the interleukin-6 (IL-6) content of mice of different groups, and c in FIG. 12 is the tumor necrosis factor- α (TNF- α) content of mice of different groups.
Fig. 13 is a graph showing H & E staining of tissue sections of different groups of mice according to the embodiment of the present invention, wherein fig. 13 a shows H & E staining of liver tissue sections of different groups of mice, and fig. 13 b shows H & E staining of epididymal adipose tissue sections of different groups of mice.
FIG. 14 is a view of the Venn diagram of a mouse intestinal microorganism species according to an embodiment of the present invention.
FIG. 15 is a bar graph (portal level) showing the abundance of intestinal microbial species in mice of different groups according to an embodiment of the present invention.
FIG. 16 is a bar graph (genus level) showing the abundance of intestinal microorganism species in mice of different groups according to an embodiment of the present invention.
Fig. 17 is an analysis of α -diversity of intestinal microorganism species of different groups of mice according to an embodiment of the present invention, wherein a in fig. 17 is the Simpson index difference of intestinal microorganisms of different groups of mice, and b in fig. 17 is the Shannon index difference of intestinal microorganisms of different groups of mice.
FIG. 18 shows the analysis of the beta-diversity of intestinal microorganism species of different groups of mice according to the present invention.
Fig. 19 is a graph showing the difference analysis of the mice intestinal microorganisms LEfSe of different groups according to the present invention.
The bifidobacterium longum subspecies infantis NKU FB3-14 of the invention is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms), and the preservation number is: CGMCC No.25762, the preservation date is 2022, 9 and 21.
Detailed Description
In order to facilitate the understanding of one bifidobacterium longum subspecies infancy NKU FB3-14 of the present invention, a more complete description of one bifidobacterium subspecies infancy NKU FB3-14 of the present invention will be given below, but without thereby limiting the scope of the present invention.
The invention relates to a bifidobacterium longum subspecies infantis NKU FB3-14:
the microbial strain is preserved in China general microbiological culture Collection center, north Chen Xili No. 1, 3 of the Korean area of Beijing city, and the preservation number is: CGMCC No.25762, the preservation date is 2022, 9 and 21.
The bifidobacterium longum subspecies infantis NKU FB3-14 have the following biological properties:
(1) Characteristics of the cells: the strain is positive in gram staining, and the strain is V-shaped, Y-shaped or club-shaped after primary or secondary culture;
(2) Colony characteristics: the colony is tiny and smooth, is milky white, has neat and raised edges and has soft texture;
(3) Growth characteristics: the strain grows anaerobically at 35-37 ℃ and enters a stationary phase after being cultured for 24-36 hours;
(4) Bifidobacterium longum subspecies infantis NKU FB3-14 is free of active nitrate reductase;
(5) Bifidobacterium longum subspecies infantis NKU FB3-14 is gamma hemolysis without hemolytic activity;
(6) The Bifidobacterium longum subspecies infantis NKU FB3-14 has acid resistance, especially in a culture environment with pH of 4+ -0.3, and the survival rate of the Bifidobacterium longum subspecies infantis NKU FB3-14 is not less than 145%;
(7) The Bifidobacterium longum subspecies infantis NKU FB3-14 has anti-bile salt property, especially in a culture environment with the mass ratio of bile salt of 0.05% +/-0.01%, the survival rate of the Bifidobacterium longum subspecies infantis NKU FB3-14 is not less than 150%;
(8) Cell culture suspension, cell-free extract and cell-free supernatant of Bifidobacterium longum subspecies infantis NKU FB3-14 have antioxidant properties, especially the clearance of cell-free supernatant to DPPH free radical is not less than 90%;
(9) In an obese mouse model, the obesity and in-vivo fat metabolism symptoms of the mouse are relieved, and bifidobacterium longum subspecies NKU FB3-14 can relieve the obesity and in-vivo fat metabolism of the mouse by inhibiting the proportion of liver and/or epididymal fat mass to the weight of the mouse;
(10) Bifidobacterium longum subspecies infantis NKU FB3-14 reduced blood glucose in mice in a mouse model of obesity to alleviate obesity and in vivo fat metabolism symptoms in mice;
(11) In an obese mouse model, bifidobacterium longum subspecies NKU FB3-14 can regulate and control the blood lipid level of the mouse, and relieve the obesity and in-vivo fat metabolism symptoms of the mouse by inhibiting the content of total cholesterol TC, total triglyceride TG and low-density lipoprotein cholesterol LDL-C and promoting the content of high-density lipoprotein cholesterol HDL-C in blood lipid;
(12) Bifidobacterium longum subspecies infantis NKU FB3-14 can regulate and control inflammatory factor relief in serum of mice, and relieve obesity and in vivo fat metabolism symptoms of the mice by reducing the content of interleukin-1β (IL-1β), interleukin-6 (IL-6) and tumor necrosis factor-alpha (TNF-alpha) in the serum;
(13) In a mouse model of obesity, bifidobacterium longum subspecies infantis NKU FB3-14 reduced high fat diet-induced histological damage by alleviating cellular steatosis, primarily by reducing expansion of adipocytes and reducing the number of adipocyte vacuoles;
(14) Bifidobacterium longum subspecies NKU FB3-14 relieves obesity and in vivo fat metabolism symptoms by regulating alpha diversity and beta diversity of intestinal flora of mice, and on a portal level, the Bifidobacterium longum subspecies NKU FB3-14 is beneficial to promoting the growth of bacterial flora of the phylum Thick-walled bacteria (Firmics), bacteroides (bacterioides) and the Desulfobacilli (Desulfobacilli) and inhibit the growth of bacterial flora of the phylum Proteus and Fusobacterium; promoting the chaetomium at the level of genus Lachnospiraceae spp.), the group NK4A136 of the genus Muril, muril @Muribaculaceaespp..Bifidobacterium genus ]Bifidibacteriumspp. and helicobacter speciesOscillospiraceaespp.) and inhibit Acinetobacter genusAcinetobacterspp.).
The extraction method of the bifidobacterium longum subspecies infantis NKU FB3-14 comprises the following steps:
isolation and screening of Bifidobacterium longum subspecies infantis NKU FB 3-14:
(1) Performing gradient dilution on the acquired infant feces sample by using sterile normal saline, performing flat plate coating on the bacterial liquid subjected to gradient dilution on a solid screening culture medium, performing anaerobic culture at 37 ℃ for 24-48 hours in a common incubator until bacterial colonies grow out, picking single bacterial colonies with bifidobacterium bacterial colony morphology, performing microscopic examination by using a gram staining method, observing the bacterial colony characteristics, and selecting gram positive bacteria as target bacterial strains.
The preparation of the solid screening medium was as follows: 50mL of the horse serum for bacteria filtration, 0.05% of L-cysteine hydrochloride and 0.05 mg/mL of mupirocin are added per liter based on the MRS culture medium, wherein the L-cysteine hydrochloride is filtered by a sterile filter membrane for sterile treatment, and the mupirocin is added when the culture medium is cooled to 40-50 ℃ after high-pressure sterilization.
(2) Purification of the strain: the strain which still has the colony morphology of the bifidobacterium after repeated streak purification on the solid screening culture medium is used as a target strain, the colony and the thallus state under a microscope are recorded, and single colony of the strain is selected to be cultured in a liquid modified MRS liquid culture Medium (MRSC) culture medium so as to prepare for subsequent molecular biological identification.
The formulation of the modified MRS liquid Medium (MRSC) is as follows: 0.05% (w/v) L-cysteine hydrochloride was added on the basis of semi-synthetic liquid MRS medium.
Colony morphology research is carried out on the strain purified by the bifidobacterium longum subspecies infantis NKU FB3-14, and the result shows that the strain colony is tiny and smooth, is milky white, has neat and raised edges and has soft texture; and the strain has V-type, Y-type or club-type shape after primary or secondary culture, as shown in figure 1.
Identification of strains: culturing the strain to mid-log phase in 37 ℃ anaerobic environment, collecting enough bacterial cells after PBS (phosphate buffered saline) washing for several times, extracting genome DNA by using a bacterial DNA extraction kit, detecting the quality of the extracted DNA, and carrying out genome sequencing and bioinformatic analysis. The phylogenetic tree was constructed by MEGA 6.0 software selection NJ (Neighbor-Joining) method by comparing with the local database, selecting the genome of 19 strains closest to bifidobacteria at the species level based on the 16S rRNA sequences, respectively. As a result, according to the analysis result, FB3-14 was confirmed to be Bifidobacterium longum, which was 99% similar to Bifidobacterium longum subspecies infancy, and further confirmed to be Bifidobacterium longum subspecies infancy, as shown in FIG. 2.
Example 1: bifidobacterium longum subspecies infantis NKU FB3-14 hemolytic assay:
strain FB3-14 was cultured to stationary phase of the third generation, streaked on a golombia blood agar plate with staphylococcus aureus ATCC25923 as a positive control, and placed in a dark place at 37 ℃ for anaerobic culture for 48 hours, and observed for the occurrence of a hemolytic circle, the transparent blood-dissolving zone appearing around the bacterial colony was β -hemolyzed, while the appearance of a green zone (α -hemolysis) or no zone (γ -hemolysis) around the bacterial colony was considered to indicate that the strain was non-hemolytic, and three replicates were made per sample. As a result, as shown in FIG. 4, it was found that Bifidobacterium longum subspecies infantis NKU FB3-14 was grown in Columbia agar plates as a non-hemolytic region, so that Bifidobacterium longum subspecies infantis NKU FB3-14 was a safe strain.
Example 2: nitrate reductase test of Bifidobacterium longum subspecies infantis NKU FB 3-14:
culturing Bifidobacterium longum subspecies NKU FB3-14 to the third generation stationary phase, inoculating to a prepared nitrate liquid culture medium by taking a blank nitrate culture medium as a negative control according to an inoculum size of 1%, adding a proper amount of sterile paraffin oil into each inoculating tube, and then placing in anaerobic culture at 37 ℃ for 48 hours; then 10 drops of 5% potassium iodide solution and 10 drops of 5% starch solution are sequentially added, vortex mixing is carried out, color change in each culture tube is observed and recorded, and three samples are parallel. As shown in FIG. 3, the bifidobacterium longum subspecies infantis NKU FB3-14 did not undergo a color reaction in the nitrate reductase activity test, and as a result, it was confirmed that the bifidobacterium longum subspecies infantis NKU FB3-14 did not contain active nitrate reductase.
Example 3: testing of acid resistance of Bifidobacterium longum subspecies infantis NKU FB 3-14:
culturing Bifidobacterium longum subspecies NKU FB3-14 to the third generation stationary phase, inoculating in MRSC liquid culture medium with pH of 3 and pH of 4 at 1% of inoculum size by volume fraction, and anaerobic culturing in anaerobic incubator at 37deg.C; respectively taking bacterial suspensions for 0 hour and 3 hours, carrying out gradient dilution, taking 200 mu L of diluted bacterial liquid on a MRSC solid culture medium flat plate, coating the flat plate by using a ball coating mode, placing the coated flat plate in an anaerobic incubator at 37 ℃ for anaerobic culture until bacterial colonies grow out, and finally counting the bacterial colonies on the flat plate. As shown in FIG. 5, it can be seen from FIG. 5 that Bifidobacterium longum subspecies NKU FB3-14 has acid resistance, and is cultured for 3 hours in an environment with pH of 4, the colony number of Bifidobacterium longum subspecies NKU FB3-14 is increased, and the survival rate of Bifidobacterium longum subspecies NKU FB3-14 is 154.88%; the survival rate of Bifidobacterium longum subspecies NKU FB3-14 was 16.22% although the colony count of Bifidobacterium longum subspecies NKU FB3-14 was reduced by 3 hours of incubation at pH 3.
Example 4: testing of bile salt tolerance of Bifidobacterium longum subspecies infantis NKU FB 3-14:
culturing Bifidobacterium longum subspecies NKU FB3-14 to third generation stationary phase, inoculating with 1% volume fraction of inoculum size in MRSC liquid culture medium with 0.05% bile salt concentration and 0.1%, and anaerobic culturing in anaerobic incubator at 37deg.C; respectively taking bacterial suspensions for 0 hour and 3 hours, carrying out gradient dilution, taking 200 mu L of diluted bacterial liquid on a plate of MRSC solid culture medium, coating the plate by using a ball coating mode, placing the coated plate in an anaerobic incubator at 37 ℃ for anaerobic culture until colonies grow out, and finally counting the plate colonies. As a result, as shown in FIG. 6, it can be seen from FIG. 6 that Bifidobacterium longum subspecies NKU FB3-14 has bile salt resistance. Wherein, in a culture environment with the mass ratio of bile salts of 0.05%, the colony number of the bifidobacterium longum subspecies infantis NKU FB3-14 is increased, and the survival rate of the bifidobacterium longum subspecies infantis NKU FB3-14 is 190.55%; bifidobacterium longum subspecies NKU FB3-14 survived when the bile salts were doubled in mass ratio.
Example 5: test of antioxidant Capacity of Bifidobacterium longum subspecies infantis NKU FB 3-14:
The 0.2mM DPPH absolute ethanol solution was mixed with the sample to be measured in a ratio of 1:1 (volume ratio), left in the dark at room temperature for 30 minutes, and then, after centrifugation at 8000rmp for 10 minutes, the absorbance of the separated supernatant was measured at 517 nm. DPPH radical scavenging activity (%) was calculated as follows:
wherein:
AS represents the absorbance of the sample;
AB represents absorbance of a blank consisting of the sample and ethanol;
AC represents absorbance of a control consisting of DPPH solution and deionized water.
Sample preparation three different samples were prepared, including cell culture suspensions of bifidobacterium longum subspecies infantis NKU FB3-14, i.e. the cell suspensions in fig. 7; cell-free extract, i.e., the cell-free extract in fig. 7; and cell-free supernatant, i.e. fermentation supernatant in FIG. 7, tested, and the test results are shown in FIG. 7. As can be seen from FIG. 7, bifidobacterium longum subspecies NKU FB3-14 have certain antioxidant properties, especially DPPH radical clearance of cell-free supernatant of Bifidobacterium longum subspecies NKU FB3-14 can be as high as 93.71%.
Example 6: remission test of bifidobacterium longum subspecies infantis NKU FB3-14 for obesity in mice:
a mouse obesity model was established and a blank, model and experimental group were set for comparison.
After placing 30C 57BL/6J mice of 4-6 weeks of age in an environment with room temperature of 20-25 ℃,12 h light/12 h dark cycle for 1 week of adaptive feeding, the mice were randomly divided into 3 groups: blank group (C), model group (M) and experiment group (T), wherein the blank group adopts normal feed, the model group adopts high-fat feed, and the experiment group adopts high-fat feed and Bifidobacterium longum subspecies infant NKU FB3-14 for intervention. During the experiment, all mice were not restricted to eating and drinking; wherein, the intervention gastric lavage dose of NKU FB3-14 of bifidobacterium longum subspecies infancy in experimental group is 10 9 CFU/mL, 200 mu L of the liquid is filled in each time, the liquid is filled continuously for 8 weeks, the liquid is weighed 1 time per week, and the feeding dosage is adjusted in time according to the weight change; the blank group and the model group are simultaneously filled with the physiological saline with the same amount as the experimental group. The change in body weight of the mice was collected during the feeding period as shown in fig. 8. Mice were given blood drawn from the rat tail tip after 16h fasting on the last day of 7-week intragastric administration and measured for fasting blood glucose. After the last gastric lavage, i.e. after modeling, all groups of mice were fasted for 12h without water, with pentobarbital sodium (40 mg/kg,1% water-soluble)Liquid, in-situ preparation) for anesthetic treatment to relieve pain of mice, collecting blood sample from orbit, standing at 4deg.C for 3 hr, centrifuging at 4deg.C for 15min at 3000 r/min, carefully sucking upper serum, packaging, and freezing in-80deg.C refrigerator. After blood taking is completed, the mice are subjected to neck breaking treatment, then liver tissues, ileum tissues and epididymal fat of the mice are immediately collected, physiological saline is soaked and then is sucked by filter paper, the cecum content of the mice is collected, and the mice are placed in a centrifuge tube and frozen by liquid nitrogen and then are frozen in a refrigerator at the temperature of minus 80 ℃ for standby. All animal experimental operations strictly follow the relevant regulations of Tianjin's regulations on experimental animal administration.
Group C mice fed the mice maintenance feed during the test period (synergistic biological SPF-grade experimental mice maintenance feed SWS9102, its main ingredients: northeast corn, wheat, imported fish meal, chicken meal, soybean oil, amino acids, vitamins, minerals); the mice in group M and group T fed high-fat nutritional feed (formula: general maintenance feed 78.8g, lard 10g, egg yolk powder 10g, cholesterol 1g, cholic acid 0.2 g) during the test period.
As can be seen from fig. 8, the initial body weight of the mice among groups is not different, and the body weight of the mice in the M groups is obviously improved by feeding the mice (model group) with high-fat feed; by continuously taking 10 9 After administration of CFU/mL doses to obese mice, and after 8 weeks of administration of the bifidobacterium longum infantis subspecies NKU FB3-14, the weight of the mice in the T group was significantly lower than those in the other two groups, and the rate of weight gain was always significantly lower in the T group than in the M group. It can be seen that Bifidobacterium longum subspecies infantis NKU FB3-14 can alleviate the weight gain of mice, thereby alleviating obesity.
Example 7: we performed the following tests on the samples obtained in example 6:
(1) Effect of bifidobacterium longum subspecies infantis NKU FB3-14 on weight and total weight of liver, epididymal fat in obese mice:
Liver tissue and epididymal fat of each sacrificed mouse were collected, and liver weight and epididymal fat weight were simultaneously weighed and recorded, and the total weight ratio of liver tissue and epididymal fat of each mouse to the body weight of the mouse was calculated, and the results are shown in fig. 9.
As can be seen from fig. 9, the mice of the model group had the highest mass ratio of liver tissue to epididymal fat, wherein the mass ratio of epididymal fat was 1.89%; the mice in the experimental group are helpful to reduce the weight of the liver and epididymal fat of the mice by perfusing the stomach with bifidobacterium longum subspecies NKU FB3-14, and the mass ratio of the epididymal fat of the mice in the experimental group is 1.58%. It can be seen that Bifidobacterium longum subspecies infantis NKU FB3-14 inhibits the weight gain of obese mice to a certain extent, and effectively relieves the obesity of the mice.
(2) Effect of bifidobacterium longum subspecies infantis NKU FB3-14 on fasting glycemia in obese mice:
mice were fasted but not watered on the last day of week 7 of gavage, the mouse pads were replaced, residual feed and mouse faeces were removed from the experimental results, and 16h later were given their tail tips to take blood, and the fasting blood glucose was measured for the different groups of mice, as shown in figure 10.
As can be seen from fig. 10, the fasting blood glucose of mice in the model group was significantly higher than that of mice in the blank group and the experimental group, and the average value was 5.47mmol/L. After 8-week-long bifidobacterium subspecies infantis NKU FB3-14 were dried in mice of the experimental group, the fasting blood glucose level of the mice was significantly reduced to 3.81mmol/L. It can be seen that intervention of Bifidobacterium longum subspecies infantis NKU FB3-14 is effective in reducing and controlling fasting blood glucose levels in mice.
(3) Effects of Bifidobacterium longum infant subspecies NKU FB3-14 on blood lipid levels in obese mice:
serum from the sacrificed mice was collected and blood lipid related index in serum from mice of different groups was determined according to commercial detection kit (institute of bioengineering, built in south kyo): total cholesterol TC, total triglycerides TG, low density lipoprotein cholesterol LDL-C and high density lipoprotein cholesterol HDL-C. The results of comparing the changes in the levels of the above indicators in the serum of mice between the different groups are shown in FIG. 11.
We can see from fig. 11 that total cholesterol TC, total triglycerides TG, low-density lipoprotein cholesterol LDL-C were significantly increased, and high-density lipoprotein cholesterol HDL-C was significantly decreased in the blood lipid related index of mice of the model group. Mice in the experimental group are closer to normal in serum by intervention of bifidobacterium longum subspecies infantis NKU FB 3-14. It can be seen that Bifidobacterium longum subspecies infancy NKU FB3-14 effectively controls and relieves dyslipidemia caused by high fat diet.
(4) Effects of Bifidobacterium longum infant subspecies NKU FB3-14 on inflammatory factors in serum of obese mice:
serum from mice was collected after sacrifice and blood lipid related index was measured in serum from mice of different groups according to commercial ELISA detection kit (institute of bioengineering, beginner, kyo): the changes in the levels of the above indicators in the serum of mice in different groups were compared with interleukin-1β (IL-1β), interleukin-6 (IL-6), and tumor necrosis factor- α (TNF- α), and the results are shown in FIG. 12.
Low-grade inflammation promotes the development of metabolic syndrome, which may lead to the development of obesity, wherein elevated levels of IL-1 beta, IL-6 and TNF-alpha lead to elevated inflammatory factors. As can be seen from FIG. 12, the characteristics include IL-1. Beta., IL-6, TNF-alpha, etc. induce a series of increases in inflammatory factors, thereby inducing metabolic inflammation. According to the experimental results in FIG. 12, the serum levels of IL-1 beta, IL-6, TNF-alpha were significantly higher in the serum of the high fat diet fed M mice than in group C, but the above inflammatory index was significantly reduced in the experimental group after the bifidobacterium subspecies infancy NKU FB3-14 had been dried. It can be seen that intervention of Bifidobacterium longum subspecies infantis NKU FB3-14 can inhibit the occurrence and development of inflammation to a certain extent, and can relieve obesity.
(5) H & E staining and pathology analysis of mouse liver tissue and epididymal adipose tissue sections:
fixing the collected part of liver tissue and epididymal fat in paraformaldehyde fixing solution, sequentially subjecting the fixed tissue to 70%, 80% and 90% ethanol solutions for 30min respectively, and then placing 95% and 100% ethanol solutions for 2 times each for 20min each time for gradient dehydration. Adding 1/2 pure alcohol, 1/2 xylene equivalent mixed solution for 15min, washing before xylene for 15min, and washing after xylene for 15min until transparent. Adding mixed solution of xylene and paraffin for 15min, and adding paraffin I and paraffin II for paraffin permeation for 50-60 min to remove the transparent agent. Embedding and slicing (slicing into 5 mu m slices with a tissue slicer), spreading, baking, H & E staining and sealing, and taking slice images with a fluorescence inverted microscope, and carrying out histopathological analysis on mice at the same time, wherein the result is shown in fig. 13.
From the liver histopathological sections of the mice of the different groups in fig. 13a, we show that the mice of group C show normal liver tissue cell morphology, no significant liver cell steatosis is observed, compared to the mice of group C, liver cell steatosis is observed in liver tissue sections of the mice of group M fed with high fat diet, a large number of lipid droplets appear, and a small inflammatory infiltration is accompanied. However, after continuous gastric lavage intervention by bifidobacterium longum subspecies infantis NKU FB3-14 (group T), the liver histopathological changes of obese mice were alleviated, and the number and volume of lipid-induced hepatocyte fat vacuoles in the diet were significantly reduced, making them approximate to normal cell morphology.
The main manifestation of obesity is an excessive increase in adipocyte size and number. FIG. 13b is an H & E stained section of epididymal fat sites of different mice, with reference to epididymal adipose tissue sections of placebo group C mice, epididymal adipose tissue sections of M mice showed significantly high fat diet induced expansion of adipocytes, whereas epididymal adipocytes of T mice, which were interfered with by Bifidobacterium longum subspecies NKU FB3-14, were closer to normal morphology. It was demonstrated that the histological damage induced by a high-fat diet could be alleviated by intake of Bifidobacterium longum subspecies infancy NKU FB 3-14.
(6) Differential comparison of flora structure in the intestinal tract of mice:
sequencing of 16S rRNA gene microorganism:
the cecal total DNA extraction was performed using the QIAamp DNA stool minikit kit (Qiagen, hilden, germany) according to the manufacturer's instructions. The total DNA content was normalized to 1 ng/. Mu.L. V3-V4 of 16S rRNA was amplified with the upstream primer 341F (5 '-CCTACGGGNGGCWGCAG-3') and the downstream primer 802R (5 '-TACNVGGGTATCTAATCC-3'). The PCR reaction procedure was as follows: 10 ng of pure DNA, 15. Mu.L of high-fidelity DNA polymerase and PCR premix, 200 nmol/L of upstream and downstream primers, water without ribozyme, and a final volume of 30. Mu.L. PCR cycling conditions included initial denaturation at 98℃for 1 min,30 cycles of 98℃for 10s,50℃for 30 s, and 72℃for 5 min. The PCR products were quantified, mixed in equimolar proportions, and then purified for sequencing using Illumina MeSeq platform, sequencing depth of at least 20000 sequences per sample.
QIIME analysis software uses a FLASH procedure to merge sequences. The double-ended sequences were pooled into one Operational Taxon (OTUs) with 97% sequence similarity using GreenGenes dataset and UPARSE algorithm. The representative sequences of each OTU are aligned and then RDP classifier is used to annotate the taxonomic information of each representative sequence. The analysis includes each taxonomic level (e.g., phylum, class, order, family, genus, etc.). In-house Perl script was used to analyze the alpha diversity within samples and beta diversity between samples. Principal coordinate analysis (PCoA) was used to evaluate the differences between experimental samples using the Unweighted unifrac method. All samples were used to calculate alpha diversity including abundance (rich) and diversity (diversity). Taxonomic information for analysis of 16S rRNA gene sequence information using UCLUST version 1.2.22 a 90% confidence interval was used for the Silva119 16S rRNA dataset. PICRUSt analysis was used to predict microbial function, and the abundance of labeled OTUs was used to clarify the highly representative flora of the different treatment groups using the LEfSe net tool.
The intestinal flora composition structure of different groups of mice was analyzed as shown in fig. 14. Of these, 2064 OTUs in the gut of group C mice, 4146 OTUs in the gut of group M mice, 731 OTUs in the gut of group T mice, but only 447 OTUs were common to the gut of group C, group M and group T mice, indicating a large change in the flora structure in the gut of the mice. It can be seen that the high fat diet affects the structure of the intestinal flora, whereas Bifidobacterium longum subspecies NKU FB3-14 stabilizes the structure of the intestinal flora in mice.
The microbial community structure of the intestinal tracts of mice of different groups was analyzed, and as shown in FIG. 15, at the portal level, the intestinal flora of mice of the blank group mainly comprises the phylum of Firmides (Firmides), bacteroides (Bacteroides), cyanobacteria (Cyanobacteria), proteus (Proteus), actinomycetes (Actinobactetaria), and Desulfobacilli (Desulfobacilli); the intestinal flora in the mice of the simulation group mainly comprises, firmicutes, bacteroides (bacterioides), cyanobacteria (Cyanobacteria), proteus (Proteobacteria), desulphurized bacillus (desulphurized), actinomycetes (actionobacteria), fusobacterium (fusobacteria). Among them, the Proteus and Fusobacterium can produce Lipopolysaccharide (LPS) and have the activity of endotoxin that causes inflammation, thereby causing the occurrence of metabolic syndrome. In the experimental group, the intestinal flora of mice in the experimental group mainly comprises the bacterial flora of the phylum Firmicutes, the bacterial flora of the pseudobacillus (bacterioides), the bacterial flora of the desulphus (desulphus) and the actinomycetes (actylobacteria), and it is seen that the bifidobacterium longum subspecies infantis NKU FB3-14 is beneficial to promote the bacterial flora growth of the Firmicutes (Firmicutes), the bacterial flora of the pseudobacillus (bacterioides) and the bacterial flora of the desulphus (desulphus) and inhibit the bacterial flora growth of the proteus and the fusobacterium.
We further studied the analysis of intestinal flora structure at the genus level in mice of different groups, as shown in figure 16. At the genus level, intervention of Bifidobacterium longum subspecies infantis NKU FB3-14 significantly increased the Chaetomium species in the mouse intestinal tractLachnospiraceae spp.), the group NK4A136 of the genus Muril, muril @Muribaculaceaespp..Bifidobacterium genus ]Bifidibacteriumspp.) abundance, and it was additionally found that helicobacter sp was positively correlated with the degree of obesity reliefOscillospiraceaespp.) there was a significant increase in the abundance of bifidobacterium longum subspecies infantis NKU FB3-14 in mice. The damage of flora composition caused by high-fat diet is also shown in Acinetobacter genusAcinetobacterspp.), but the bacterial abundance is inhibited in group T mice. The above results indicate that intervention by bifidobacterium longum subspecies infantis NKU FB3-14 significantly reversed the abnormal expression of specific bacteria induced by the high fat diet, promoting the sustained enrichment of beneficial bacteria.
We further investigated the effect of Bifidobacterium longum subspecies infantis NKU FB3-14 on the abundance and alpha diversity of the flora in the intestinal flora of mice, as shown in FIG. 17. The Shannon index and the Simpson index are used for measuring the species diversity of the intestinal microorganisms of the mice, and an experimental group shows a remarkably higher index value, namely, after the bifidobacterium longum subspecies NKU FB3-14 are dried, the uniformity of each species in the intestinal microorganism community of the mice is remarkably higher, the species diversity is more, and the dysbacteriosis of obese mice is regulated to a certain extent, so that the obesity is relieved.
We further used principal component analysis PCA (Principal Component Analysis) for phylogenetic difference analysis (β diversity) of intestinal microorganisms as shown in fig. 18. As can be seen from fig. 18, the results of clustering analysis of intestinal microbiota of mice in the same group showed that the contribution of the first principal component to the sample difference was 37.64% and the contribution of the second principal component to the sample difference was 28.22%, and that bifidobacterium longum subspecies infantis NKU FB3-14 had an effect on the β -diversity of the intestinal microbiota while alleviating obesity.
To identify related microorganisms that were treated by Bifidobacterium longum subspecies infantis NKU FB3-14 to improve metabolic conditions, we compared the intestinal microbiota of the different treatment groups using the LEfSe assay. LEfSe revealed differences in the flora of each group from phylum to species level (fig. 19). LEfSe analysis showed that a total of 48 OTUs were screened as phylogenetic types, whose relative abundance varied significantly due to high fat diet feeding and bifidobacterium longum subspecies infancy NKU FB3-14 treatment. In the body (model group) of mice fed by high-fat diet, pathogenic taxonomic groups such as Proteus, acinetobacter, fusobacterium and the like are relatively enriched, and have obvious differential expression; while some beneficial flora, such as those of the phylum firmicutes, the genus helicobacter, the genus chaetomium, NK4a136, were observed to be more abundant in the experimental group, showing significant differences from the other groups. The results show that Bifidobacterium longum subspecies infantis NKU FB3-14 inhibits the detrimental flora in the intestinal tract and improves the flora structure of the intestinal microorganisms.
The 16s DNA sequence of the bifidobacterium longum subspecies infantis NKU FB3-14 is shown as follows:
tcctgggggt ctaccatgca gtcgaacggg atccatcaag cttgcttggt ggtgagagtg 60 gcgaacgggt gagtaatgcg tgaccgacct gccccataca ccggaatagc tcctggaaac 120 gggtggtaat gccggatgtt ccagttgatc gcatggtctt ctgggaaagc tttcgcggta 180 tgggatgggg tcgcgtccta tcagcttgac ggcggggtaa cggcccaccg tggcttcgac 240 gggtagccgg cctgagaggg cgaccggcca cattgggact gagatacggc ccagactcct 300 acgggaggca gcagtgggga atattgcaca atgggcgcaa gcctgatgca gcgacgccgc 360 gtgagggatg gaggccttcg ggttgtaaac ctcttttatc ggggagcaag cgtgagtgag 420 tttacccgtt gaataagcac cggctaacta cgtgccagca gccgcggtaa tacgtagggt 480 gcaagcgtta tccggaatta ttgggcgtaa agggctcgta ggcggttcgt cgcgtccggt 540 gtgaaagtcc atcgcttaac ggtggatccg cgccgggtac gggcgggctt gagtgcggta 600 ggggagactg gaattcccgg tgtaacggtg gaatgtgtag atatcgggaa gaacaccaat 660 ggcgaaggca ggtctctggg ccgttactga cgctgaggag cgaaagcgtg gggagcgaac 720 aggattagat accctggtag tccacgccgt aaacggtgga tgctggatgt ggggcccgtt 780 ccacgggttc cgtgtcggag ctaacgcgtt aagcatcccg cctggggagt acggccgcaa 840 ggctaaaact caaagaaatt gacgggggcc cgcacaagcg gcggagcatg cggattaatt 900 cgatgcaacg cgaagaacct tacctgggct tgacatgttc ccgacgatcc cagagatggg 960 gtttcccttc ggggcgggtt cacaggtggt gcatggtcgt cgtcagctcg tgtcgtgaga 1020 tgttgggtta agtcccgcaa cgagcgcaac cctcgccccg tgttgccagc ggattgtgcc 1080 gggaactcac gggggaccgc cggggttaac tcggaggaag gtggggatga cgtcagatca 1140 tcatgcccct tacgtccagg gcttcacgca tgctacaatg gccggtacaa cgggatgcga 1200 cgcggcgacg cggagcggat ccctgaaaac cggtctcagt tcggatcgca gtctgcaact 1260 cgactgcgtg aaggcggagt cgctagtaat cgcgaatcag caacgtcgcg gtgaatgcgt 1320 tcccgggcct tgtacacacc gcccgtcaag tcatgaaagt gggcagcacc cgaagccggt 1380 ggcctaaccc cttgtgggat ggagccgtct aagtagcccg gcatggcg 1428
wherein, the 33 th to 64 th bits of the 16s DNA of the bifidobacterium longum subspecies infancy NKU FB3-14 are the variable region V1, the 104 th to 208 th bits are the variable region V2, the 400 th to 443 th bits are the variable region V3, the 523 th to 628 th bits are the variable region V4, the 769 th to 827 th bits are the variable region V5, the 936 th to 991 th bits are the variable region V6, the 1066 th to 1122 th bits are the variable region V7, the 1193 th to 1243 th bits are the variable region V8, and the 1386 th to 1428 th bits are the variable region V9.
In conclusion, the bifidobacterium longum subspecies infantis NKU FB3-14 of the invention can be used for preparing functional foods or medicines for obesity and/or regulating in-vivo fat metabolism.
It should be noted that the above-mentioned embodiments are only preferred embodiments of the present invention, and are not intended to limit the present invention, and any modifications, equivalent substitutions, improvements, etc. within the spirit and principle of the present invention should be included in the protection scope of the present invention.
Claims (9)
1. Use of bifidobacterium longum subspecies infantis NKUFB3-14 deposited with the chinese microbiological bacterial strain deposit management committee common microbiological centre, no. 3 of north western road 1, the region of facing yang in beijing, deposit No. 3 of the national institutes of microbiological bacterial deposit management: CGMCC No.25762, the preservation date is 2022, 9 and 21.
2. The use according to claim 1, wherein the preparation of a medicament for alleviating obesity and/or regulating fat metabolism in the body is a medicament having at least one of the following actions:
i) a pharmaceutical product having a weight-on-weight-reducing effect on liver and/or epididymal fat mass;
II) a medicament having a blood glucose regulating effect;
III) drugs with the ability to modulate the levels of inflammatory factors in serum;
IV) a medicament for regulating blood lipid metabolic disorders;
v) a drug having an effect of alleviating high-fat diet-induced liver histological damage;
VI) a pharmaceutical product with a structure for regulating the high fat metabolizing intestinal flora;
the III) has the medicine for regulating the level of inflammatory factors in serum, in particular to the medicine for reducing the contents of interleukin-1 beta, interleukin-6 and tumor necrosis factor-alpha in serum.
3. Use according to claim 2, characterized in that iv) has a pharmaceutical for regulating the metabolic disorders of blood lipids, in particular by inhibiting the content of total cholesterol TC, total triglycerides TG, low-density lipoprotein cholesterol LDL-C in blood lipids and promoting the content of high-density lipoprotein cholesterol HDL-C in blood lipids.
4. Use according to claim 2, wherein v) has a drug for alleviating high-fat diet-induced liver histological damage, in particular a drug for alleviating hepatic cell steatosis.
5. The use according to claim 4, wherein the medicament with effect of alleviating hepatic steatosis, in particular by reducing the expansion of adipocytes and reducing the number of adipocyte vacuoles.
6. Use according to claim 2, wherein said vi) drug with structure regulating the high lipid metabolizing intestinal flora is a drug with the following effects:
i) drugs that modulate the alpha-or beta-diversity of the intestinal flora of mice.
7. Use according to claim 6, wherein said drug substance is inhibited by modulating β -diversity of the mouse intestinal flora, in particular by promoting growth of the flora of the phylum firmicutes, bacteroidetes, desulphus, fusobacterium, at the phylum of the phylum; at the genus level, growth of the genus chaetobacter (lachnospiracepp), the group NK4a136 of the genus chaetobacter, the genus Muri (muraculolacearpp), the genus bifidobacterium (bifidum spp.), and the genus helicobacter (oscillascopepast esppp) is promoted, and growth of the genus acinetobacter (acinetobacter spp) is suppressed.
8. The application of Bifidobacterium longum subspecies infantis NKUFB3-14 in preparing health food, wherein Bifidobacterium longum subsp.inffantis is preserved in China general microbiological culture Collection center (China Committee for culture Collection of microorganisms), north Xielu No. 1, no. 3 in the area of towards the sun in Beijing city, with the preservation number: cgmccno.25762, collection date 2022, 9 and 21;
The health food is at least one of the following health food:
i) health food which helps to control fat in the body;
II) health food for maintaining blood lipid;
III) health food for helping to maintain blood sugar.
9. The use according to claim 8, characterized in that the health food which contributes to the maintenance of blood lipids, in particular by inhibiting the content of total cholesterol TC, total triglycerides TG, low-density lipoprotein cholesterol LDL-C in blood lipids and promoting the content of high-density lipoprotein cholesterol HDL-C in blood lipids.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310453211.XA CN116286551B (en) | 2023-04-25 | 2023-04-25 | Application of bifidobacterium longum subspecies infantis in regulating in-vivo fat metabolism, shaping, reducing fat and improving obesity |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202310453211.XA CN116286551B (en) | 2023-04-25 | 2023-04-25 | Application of bifidobacterium longum subspecies infantis in regulating in-vivo fat metabolism, shaping, reducing fat and improving obesity |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116286551A CN116286551A (en) | 2023-06-23 |
| CN116286551B true CN116286551B (en) | 2023-08-11 |
Family
ID=86803363
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202310453211.XA Active CN116286551B (en) | 2023-04-25 | 2023-04-25 | Application of bifidobacterium longum subspecies infantis in regulating in-vivo fat metabolism, shaping, reducing fat and improving obesity |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116286551B (en) |
Families Citing this family (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| 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 |
| CN117210380B (en) * | 2023-11-09 | 2024-01-26 | 天赋能(天津)功能食品研究发展有限公司 | Application of bifidobacterium longum subspecies infantis NKU FB3-14 in tumor inhibition |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2526986A1 (en) * | 2013-07-18 | 2015-01-19 | Biopolis, S.L. | Novel strain of bifidobacterium animalis subsp. lactis cect 8145 and use thereof for the treatment and/or prevention of excess weight and obesity and associated diseases |
| CN115927049A (en) * | 2022-07-20 | 2023-04-07 | 华南理工大学 | Bifidobacterium longum subspecies infantis B2-01 and application thereof |
-
2023
- 2023-04-25 CN CN202310453211.XA patent/CN116286551B/en active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| ES2526986A1 (en) * | 2013-07-18 | 2015-01-19 | Biopolis, S.L. | Novel strain of bifidobacterium animalis subsp. lactis cect 8145 and use thereof for the treatment and/or prevention of excess weight and obesity and associated diseases |
| CN115927049A (en) * | 2022-07-20 | 2023-04-07 | 华南理工大学 | Bifidobacterium longum subspecies infantis B2-01 and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116286551A (en) | 2023-06-23 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN116286551B (en) | Application of bifidobacterium longum subspecies infantis in regulating in-vivo fat metabolism, shaping, reducing fat and improving obesity | |
| CN112501046B (en) | Lactobacillus fermentum with weight-losing function and application thereof | |
| CN108076643B (en) | Lactobacillus rhamnosus bacteria for the treatment of e.g. bacterial vaginosis | |
| CN111254090B (en) | Lactobacillus reuteri with weight losing function and application thereof | |
| JP6630388B2 (en) | Lactobacillus plantarum TCI378 and its use in reducing fat and improving gastrointestinal function | |
| CN114107121B (en) | Bacillus coagulans and application thereof in treatment of alcoholic liver disease | |
| WO2019227418A1 (en) | Composition and uses thereof | |
| CN114766677A (en) | Application of lactobacillus rhamnosus LRa05 in preparing preparation for improving irritable bowel syndrome | |
| US20130309212A1 (en) | Lactobacillus salivarius and method for preparing metabolite thereof, composition of lactobacillus salivarius and metabolite thereof and use of the composition | |
| CN114657084B (en) | Bifidobacterium longum for relieving ulcerative colitis and application thereof | |
| CN113512514B (en) | Lactococcus lactis with depression improving effect and application thereof | |
| CN113575758A (en) | Composite probiotics and preparation method thereof | |
| CN116121154B (en) | Leuconostoc lactis and application thereof | |
| CN109576165B (en) | Saccharomyces bayanus and application thereof | |
| CN116445356A (en) | Bifidobacterium animalis subspecies BA67 for regulating intestinal flora and enhancing immunity and application thereof | |
| CN114686405B (en) | Bifidobacterium bifidum with functions of reducing fat, relieving hyperglycemia and regulating intestinal immunity and application thereof | |
| CN114933992B (en) | Application of bifidobacterium longum and composite preparation thereof in relieving ulcerative colitis | |
| CN116555102A (en) | Lactobacillus plantarum producing gamma-aminobutyric acid, sleep-aiding probiotic composition and preparation method thereof | |
| WO2019227414A1 (en) | Composition and uses thereof | |
| CN115992059A (en) | Lactobacillus johnsonii for producing feruloyl esterase and application thereof in relieving ulcerative colitis | |
| WO2019227417A1 (en) | Composition and uses thereof | |
| CN115261251B (en) | Streptococcus thermophilus S869 and application thereof in regulating immunity and intestinal functions | |
| CN116694534A (en) | Bifidobacterium longum SX-1326 and application thereof | |
| CN115074277A (en) | Clostridium pralatanorum capable of relieving obesity and application thereof | |
| CN116004472A (en) | Clostridium butyricum for relieving obesity and application 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 | ||
| GR01 | Patent grant | ||
| GR01 | Patent grant |