CN110964659A - Lactobacillus plantarum with blood sugar reducing effect, screening method thereof, application medicament and food - Google Patents
Lactobacillus plantarum with blood sugar reducing effect, screening method thereof, application medicament and food Download PDFInfo
- Publication number
- CN110964659A CN110964659A CN201910709658.2A CN201910709658A CN110964659A CN 110964659 A CN110964659 A CN 110964659A CN 201910709658 A CN201910709658 A CN 201910709658A CN 110964659 A CN110964659 A CN 110964659A
- Authority
- CN
- China
- Prior art keywords
- cells
- lactic acid
- lactobacillus plantarum
- acid bacteria
- solution
- 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
- 238000000034 method Methods 0.000 title claims abstract description 42
- 240000006024 Lactobacillus plantarum Species 0.000 title claims abstract description 36
- 235000013965 Lactobacillus plantarum Nutrition 0.000 title claims abstract description 36
- 229940072205 lactobacillus plantarum Drugs 0.000 title claims abstract description 36
- 238000012216 screening Methods 0.000 title claims abstract description 33
- 239000008280 blood Substances 0.000 title claims abstract description 27
- 210000004369 blood Anatomy 0.000 title claims abstract description 27
- 230000001603 reducing effect Effects 0.000 title claims abstract description 24
- 235000000346 sugar Nutrition 0.000 title claims abstract description 23
- 239000003814 drug Substances 0.000 title claims abstract description 10
- 235000013305 food Nutrition 0.000 title claims description 3
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 claims abstract description 114
- 241000894006 Bacteria Species 0.000 claims abstract description 70
- 239000004310 lactic acid Substances 0.000 claims abstract description 57
- 235000014655 lactic acid Nutrition 0.000 claims abstract description 57
- 230000001580 bacterial effect Effects 0.000 claims abstract description 21
- 230000000694 effects Effects 0.000 claims abstract description 15
- 238000002474 experimental method Methods 0.000 claims abstract description 12
- 239000006041 probiotic Substances 0.000 claims abstract description 10
- 235000018291 probiotics Nutrition 0.000 claims abstract description 10
- 238000001727 in vivo Methods 0.000 claims abstract description 9
- 238000004321 preservation Methods 0.000 claims abstract description 9
- 230000000529 probiotic effect Effects 0.000 claims abstract description 8
- 230000003178 anti-diabetic effect Effects 0.000 claims abstract description 7
- 238000012795 verification Methods 0.000 claims abstract description 6
- 230000003647 oxidation Effects 0.000 claims abstract description 3
- 238000007254 oxidation reaction Methods 0.000 claims abstract description 3
- 239000000243 solution Substances 0.000 claims description 32
- 239000000523 sample Substances 0.000 claims description 28
- 241000700159 Rattus Species 0.000 claims description 27
- 239000001963 growth medium Substances 0.000 claims description 21
- 241000186660 Lactobacillus Species 0.000 claims description 20
- 229940039696 lactobacillus Drugs 0.000 claims description 20
- 238000006243 chemical reaction Methods 0.000 claims description 17
- 239000006228 supernatant Substances 0.000 claims description 17
- 239000003833 bile salt Substances 0.000 claims description 16
- 239000012530 fluid Substances 0.000 claims description 16
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 claims description 16
- 238000012258 culturing Methods 0.000 claims description 15
- 230000002496 gastric effect Effects 0.000 claims description 14
- 239000000725 suspension Substances 0.000 claims description 13
- 238000000855 fermentation Methods 0.000 claims description 12
- 230000004151 fermentation Effects 0.000 claims description 12
- 208000001072 type 2 diabetes mellitus Diseases 0.000 claims description 12
- 238000002835 absorbance Methods 0.000 claims description 11
- 239000000284 extract Substances 0.000 claims description 11
- 230000031700 light absorption Effects 0.000 claims description 11
- 206010012601 diabetes mellitus Diseases 0.000 claims description 10
- 239000008103 glucose Substances 0.000 claims description 10
- 238000002156 mixing Methods 0.000 claims description 10
- 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 9
- 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 claims description 9
- 230000005764 inhibitory process Effects 0.000 claims description 9
- 230000004083 survival effect Effects 0.000 claims description 9
- 102100023374 Forkhead box protein M1 Human genes 0.000 claims description 8
- 101000907578 Homo sapiens Forkhead box protein M1 Proteins 0.000 claims description 8
- 102100024295 Maltase-glucoamylase Human genes 0.000 claims description 8
- 108010028144 alpha-Glucosidases Proteins 0.000 claims description 8
- 230000003859 lipid peroxidation Effects 0.000 claims description 8
- 239000007788 liquid Substances 0.000 claims description 8
- 235000004213 low-fat Nutrition 0.000 claims description 8
- 238000005259 measurement Methods 0.000 claims description 8
- 239000000203 mixture Substances 0.000 claims description 8
- 108090000790 Enzymes Proteins 0.000 claims description 7
- 102000004190 Enzymes Human genes 0.000 claims description 7
- 229940088598 enzyme Drugs 0.000 claims description 7
- 239000002609 medium Substances 0.000 claims description 7
- 239000007787 solid Substances 0.000 claims description 7
- 238000005406 washing Methods 0.000 claims description 7
- TUJKJAMUKRIRHC-UHFFFAOYSA-N hydroxyl Chemical compound [OH] TUJKJAMUKRIRHC-UHFFFAOYSA-N 0.000 claims description 6
- 230000003914 insulin secretion Effects 0.000 claims description 6
- 239000002504 physiological saline solution Substances 0.000 claims description 6
- ZSJLQEPLLKMAKR-GKHCUFPYSA-N streptozocin Chemical compound O=NN(C)C(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@@H](O)[C@@H]1O ZSJLQEPLLKMAKR-GKHCUFPYSA-N 0.000 claims description 6
- 238000002965 ELISA Methods 0.000 claims description 5
- 239000003472 antidiabetic agent Substances 0.000 claims description 5
- 239000003963 antioxidant agent Substances 0.000 claims description 5
- 230000003078 antioxidant effect Effects 0.000 claims description 5
- 235000006708 antioxidants Nutrition 0.000 claims description 5
- 238000001514 detection method Methods 0.000 claims description 5
- 210000001035 gastrointestinal tract Anatomy 0.000 claims description 5
- 230000000968 intestinal effect Effects 0.000 claims description 5
- 210000004153 islets of langerhan Anatomy 0.000 claims description 5
- 238000009630 liquid culture Methods 0.000 claims description 5
- CIWBSHSKHKDKBQ-JLAZNSOCSA-N Ascorbic acid Chemical compound OC[C@H](O)[C@H]1OC(=O)C(O)=C1O CIWBSHSKHKDKBQ-JLAZNSOCSA-N 0.000 claims description 4
- -1 DPPH free radical Chemical class 0.000 claims description 4
- 239000006144 Dulbecco’s modified Eagle's medium Substances 0.000 claims description 4
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 4
- 241001465754 Metazoa Species 0.000 claims description 4
- 230000003115 biocidal effect Effects 0.000 claims description 4
- 239000007853 buffer solution Substances 0.000 claims description 4
- 238000003304 gavage Methods 0.000 claims description 4
- 230000015784 hyperosmotic salinity response Effects 0.000 claims description 4
- 239000000843 powder Substances 0.000 claims description 4
- 238000012360 testing method Methods 0.000 claims description 4
- YNJBWRMUSHSURL-UHFFFAOYSA-N trichloroacetic acid Chemical compound OC(=O)C(Cl)(Cl)Cl YNJBWRMUSHSURL-UHFFFAOYSA-N 0.000 claims description 4
- 108010019160 Pancreatin Proteins 0.000 claims description 3
- 238000004113 cell culture Methods 0.000 claims description 3
- 238000010276 construction Methods 0.000 claims description 3
- 235000009200 high fat diet Nutrition 0.000 claims description 3
- 230000002401 inhibitory effect Effects 0.000 claims description 3
- 210000000936 intestine Anatomy 0.000 claims description 3
- 229940055695 pancreatin Drugs 0.000 claims description 3
- 238000005502 peroxidation Methods 0.000 claims description 3
- RVBUGGBMJDPOST-UHFFFAOYSA-N 2-thiobarbituric acid Chemical compound O=C1CC(=O)NC(=S)N1 RVBUGGBMJDPOST-UHFFFAOYSA-N 0.000 claims description 2
- BTJIUGUIPKRLHP-UHFFFAOYSA-N 4-nitrophenol Chemical compound OC1=CC=C([N+]([O-])=O)C=C1 BTJIUGUIPKRLHP-UHFFFAOYSA-N 0.000 claims description 2
- IFBHRQDFSNCLOZ-IIRVCBMXSA-N 4-nitrophenyl-α-d-galactoside Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1OC1=CC=C([N+]([O-])=O)C=C1 IFBHRQDFSNCLOZ-IIRVCBMXSA-N 0.000 claims description 2
- 108091003079 Bovine Serum Albumin Proteins 0.000 claims description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 claims description 2
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 claims description 2
- ZDXPYRJPNDTMRX-VKHMYHEASA-N L-glutamine Chemical compound OC(=O)[C@@H](N)CCC(N)=O ZDXPYRJPNDTMRX-VKHMYHEASA-N 0.000 claims description 2
- 229930182816 L-glutamine Natural products 0.000 claims description 2
- OYHQOLUKZRVURQ-HZJYTTRNSA-N Linoleic acid Chemical compound CCCCC\C=C/C\C=C/CCCCCCCC(O)=O OYHQOLUKZRVURQ-HZJYTTRNSA-N 0.000 claims description 2
- 241000700157 Rattus norvegicus Species 0.000 claims description 2
- ZSJLQEPLLKMAKR-UHFFFAOYSA-N Streptozotocin Natural products O=NN(C)C(=O)NC1C(O)OC(CO)C(O)C1O ZSJLQEPLLKMAKR-UHFFFAOYSA-N 0.000 claims description 2
- 230000009471 action Effects 0.000 claims description 2
- 230000003698 anagen phase Effects 0.000 claims description 2
- 235000010323 ascorbic acid Nutrition 0.000 claims description 2
- 229960005070 ascorbic acid Drugs 0.000 claims description 2
- 239000011668 ascorbic acid Substances 0.000 claims description 2
- 238000004364 calculation method Methods 0.000 claims description 2
- 239000013553 cell monolayer Substances 0.000 claims description 2
- 230000004663 cell proliferation Effects 0.000 claims description 2
- 238000005119 centrifugation Methods 0.000 claims description 2
- 229960001701 chloroform Drugs 0.000 claims description 2
- 238000001816 cooling Methods 0.000 claims description 2
- XUJNEKJLAYXESH-UHFFFAOYSA-N cysteine Natural products SCC(N)C(O)=O XUJNEKJLAYXESH-UHFFFAOYSA-N 0.000 claims description 2
- 235000018417 cysteine Nutrition 0.000 claims description 2
- 239000012153 distilled water Substances 0.000 claims description 2
- 239000000839 emulsion Substances 0.000 claims description 2
- 230000003203 everyday effect Effects 0.000 claims description 2
- 238000000605 extraction Methods 0.000 claims description 2
- 239000011790 ferrous sulphate Substances 0.000 claims description 2
- 235000003891 ferrous sulphate Nutrition 0.000 claims description 2
- 239000012091 fetal bovine serum Substances 0.000 claims description 2
- 230000006870 function Effects 0.000 claims description 2
- 230000012010 growth Effects 0.000 claims description 2
- BAUYGSIQEAFULO-UHFFFAOYSA-L iron(2+) sulfate (anhydrous) Chemical compound [Fe+2].[O-]S([O-])(=O)=O BAUYGSIQEAFULO-UHFFFAOYSA-L 0.000 claims description 2
- 229910000359 iron(II) sulfate Inorganic materials 0.000 claims description 2
- OYHQOLUKZRVURQ-IXWMQOLASA-N linoleic acid Natural products CCCCC\C=C/C\C=C\CCCCCCCC(O)=O OYHQOLUKZRVURQ-IXWMQOLASA-N 0.000 claims description 2
- 235000020778 linoleic acid Nutrition 0.000 claims description 2
- 238000000465 moulding Methods 0.000 claims description 2
- 239000002244 precipitate Substances 0.000 claims description 2
- 239000012488 sample solution Substances 0.000 claims description 2
- CDBYLPFSWZWCQE-UHFFFAOYSA-L sodium carbonate Substances [Na+].[Na+].[O-]C([O-])=O CDBYLPFSWZWCQE-UHFFFAOYSA-L 0.000 claims description 2
- 229910000029 sodium carbonate Inorganic materials 0.000 claims description 2
- GNBVPFITFYNRCN-UHFFFAOYSA-M sodium thioglycolate Chemical compound [Na+].[O-]C(=O)CS GNBVPFITFYNRCN-UHFFFAOYSA-M 0.000 claims description 2
- 229940046307 sodium thioglycolate Drugs 0.000 claims description 2
- 239000008223 sterile water Substances 0.000 claims description 2
- 230000004936 stimulating effect Effects 0.000 claims description 2
- 229960001052 streptozocin Drugs 0.000 claims description 2
- 239000000126 substance Substances 0.000 claims description 2
- 230000035899 viability Effects 0.000 claims description 2
- 230000000638 stimulation Effects 0.000 claims 2
- XPFJYKARVSSRHE-UHFFFAOYSA-K trisodium;2-hydroxypropane-1,2,3-tricarboxylate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].[Na+].[Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.[O-]C(=O)CC(O)(CC([O-])=O)C([O-])=O XPFJYKARVSSRHE-UHFFFAOYSA-K 0.000 claims 2
- 101100191768 Caenorhabditis elegans pbs-4 gene Proteins 0.000 claims 1
- 241001052560 Thallis Species 0.000 claims 1
- 238000003287 bathing Methods 0.000 claims 1
- 230000007910 cell fusion Effects 0.000 claims 1
- 239000012295 chemical reaction liquid Substances 0.000 claims 1
- 238000007865 diluting Methods 0.000 claims 1
- 235000020776 essential amino acid Nutrition 0.000 claims 1
- 239000003797 essential amino acid Substances 0.000 claims 1
- 230000002779 inactivation Effects 0.000 claims 1
- 238000002372 labelling Methods 0.000 claims 1
- 239000011259 mixed solution Substances 0.000 claims 1
- 239000008055 phosphate buffer solution Substances 0.000 claims 1
- 238000000338 in vitro Methods 0.000 abstract description 7
- 239000002953 phosphate buffered saline Substances 0.000 description 11
- 230000002218 hypoglycaemic effect Effects 0.000 description 9
- 238000011160 research Methods 0.000 description 7
- 206010022489 Insulin Resistance Diseases 0.000 description 6
- 238000000513 principal component analysis Methods 0.000 description 5
- ULGZDMOVFRHVEP-RWJQBGPGSA-N Erythromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@@](C)(O)[C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 ULGZDMOVFRHVEP-RWJQBGPGSA-N 0.000 description 4
- 239000003223 protective agent Substances 0.000 description 4
- 230000002000 scavenging effect Effects 0.000 description 4
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 description 3
- 230000002292 Radical scavenging effect Effects 0.000 description 3
- 239000003242 anti bacterial agent Substances 0.000 description 3
- 229940088710 antibiotic agent Drugs 0.000 description 3
- 229940079593 drug Drugs 0.000 description 3
- HDTRYLNUVZCQOY-UHFFFAOYSA-N α-D-glucopyranosyl-α-D-glucopyranoside Natural products OC1C(O)C(O)C(CO)OC1OC1C(O)C(O)C(O)C(CO)O1 HDTRYLNUVZCQOY-UHFFFAOYSA-N 0.000 description 2
- WQZGKKKJIJFFOK-SVZMEOIVSA-N (+)-Galactose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@H]1O WQZGKKKJIJFFOK-SVZMEOIVSA-N 0.000 description 2
- VTYYLEPIZMXCLO-UHFFFAOYSA-L Calcium carbonate Chemical compound [Ca+2].[O-]C([O-])=O VTYYLEPIZMXCLO-UHFFFAOYSA-L 0.000 description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 description 2
- WHUUTDBJXJRKMK-VKHMYHEASA-N L-glutamic acid Chemical compound OC(=O)[C@@H](N)CCC(O)=O WHUUTDBJXJRKMK-VKHMYHEASA-N 0.000 description 2
- 229930195725 Mannitol Natural products 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- DRTQHJPVMGBUCF-XVFCMESISA-N Uridine Chemical compound O[C@@H]1[C@H](O)[C@@H](CO)O[C@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-XVFCMESISA-N 0.000 description 2
- 108010059993 Vancomycin Proteins 0.000 description 2
- PYMYPHUHKUWMLA-LMVFSUKVSA-N aldehydo-D-ribose Chemical compound OC[C@@H](O)[C@@H](O)[C@@H](O)C=O PYMYPHUHKUWMLA-LMVFSUKVSA-N 0.000 description 2
- HDTRYLNUVZCQOY-LIZSDCNHSA-N alpha,alpha-trehalose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-LIZSDCNHSA-N 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- SRBFZHDQGSBBOR-UHFFFAOYSA-N beta-D-Pyranose-Lyxose Natural products OC1COC(O)C(O)C1O SRBFZHDQGSBBOR-UHFFFAOYSA-N 0.000 description 2
- 239000000872 buffer Substances 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 229960005091 chloramphenicol Drugs 0.000 description 2
- 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 2
- 229940099352 cholate Drugs 0.000 description 2
- 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 2
- 229960002227 clindamycin Drugs 0.000 description 2
- KDLRVYVGXIQJDK-AWPVFWJPSA-N clindamycin Chemical compound CN1C[C@H](CCC)C[C@H]1C(=O)N[C@H]([C@H](C)Cl)[C@@H]1[C@H](O)[C@H](O)[C@@H](O)[C@@H](SC)O1 KDLRVYVGXIQJDK-AWPVFWJPSA-N 0.000 description 2
- 229960003276 erythromycin Drugs 0.000 description 2
- 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 2
- 239000000594 mannitol Substances 0.000 description 2
- 235000010355 mannitol Nutrition 0.000 description 2
- 230000007246 mechanism Effects 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000008569 process Effects 0.000 description 2
- 150000003254 radicals Chemical class 0.000 description 2
- 230000009467 reduction Effects 0.000 description 2
- JQXXHWHPUNPDRT-WLSIYKJHSA-N rifampicin Chemical compound O([C@](C1=O)(C)O/C=C/[C@@H]([C@H]([C@@H](OC(C)=O)[C@H](C)[C@H](O)[C@H](C)[C@@H](O)[C@@H](C)\C=C\C=C(C)/C(=O)NC=2C(O)=C3C([O-])=C4C)C)OC)C4=C1C3=C(O)C=2\C=N\N1CC[NH+](C)CC1 JQXXHWHPUNPDRT-WLSIYKJHSA-N 0.000 description 2
- 229960001225 rifampicin Drugs 0.000 description 2
- 230000035945 sensitivity Effects 0.000 description 2
- 239000011780 sodium chloride Substances 0.000 description 2
- OESFSXYRSCBAQJ-UHFFFAOYSA-M sodium;3-carboxy-3,5-dihydroxy-5-oxopentanoate;2-hydroxypropane-1,2,3-tricarboxylic acid Chemical compound [Na+].OC(=O)CC(O)(C(O)=O)CC(O)=O.OC(=O)CC(O)(C(O)=O)CC([O-])=O OESFSXYRSCBAQJ-UHFFFAOYSA-M 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
- 229960003165 vancomycin Drugs 0.000 description 2
- 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 2
- 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 2
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 description 1
- 206010067484 Adverse reaction Diseases 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- GUBGYTABKSRVRQ-UHFFFAOYSA-N D-Cellobiose Natural products OCC1OC(OC2C(O)C(O)C(O)OC2CO)C(O)C(O)C1O GUBGYTABKSRVRQ-UHFFFAOYSA-N 0.000 description 1
- GUBGYTABKSRVRQ-CUHNMECISA-N D-Cellobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-CUHNMECISA-N 0.000 description 1
- RFSUNEUAIZKAJO-VRPWFDPXSA-N D-Fructose Natural products OC[C@H]1OC(O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-VRPWFDPXSA-N 0.000 description 1
- FBPFZTCFMRRESA-FSIIMWSLSA-N D-Glucitol Natural products OC[C@H](O)[C@H](O)[C@@H](O)[C@H](O)CO FBPFZTCFMRRESA-FSIIMWSLSA-N 0.000 description 1
- FBPFZTCFMRRESA-JGWLITMVSA-N D-glucitol Chemical compound OC[C@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-JGWLITMVSA-N 0.000 description 1
- WQZGKKKJIJFFOK-QTVWNMPRSA-N D-mannopyranose Chemical compound OC[C@H]1OC(O)[C@@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-QTVWNMPRSA-N 0.000 description 1
- SRBFZHDQGSBBOR-IOVATXLUSA-N D-xylopyranose Chemical compound O[C@@H]1COC(O)[C@H](O)[C@H]1O SRBFZHDQGSBBOR-IOVATXLUSA-N 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 206010059866 Drug resistance Diseases 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- RFSUNEUAIZKAJO-ARQDHWQXSA-N Fructose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O RFSUNEUAIZKAJO-ARQDHWQXSA-N 0.000 description 1
- 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
- 102000006587 Glutathione peroxidase Human genes 0.000 description 1
- 108700016172 Glutathione peroxidases Proteins 0.000 description 1
- 102000004877 Insulin Human genes 0.000 description 1
- 108090001061 Insulin Proteins 0.000 description 1
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 description 1
- 229920000057 Mannan Polymers 0.000 description 1
- OVRNDRQMDRJTHS-CBQIKETKSA-N N-Acetyl-D-Galactosamine Chemical compound CC(=O)N[C@H]1[C@@H](O)O[C@H](CO)[C@H](O)[C@@H]1O OVRNDRQMDRJTHS-CBQIKETKSA-N 0.000 description 1
- MBLBDJOUHNCFQT-UHFFFAOYSA-N N-acetyl-D-galactosamine Natural products CC(=O)NC(C=O)C(O)C(O)C(O)CO MBLBDJOUHNCFQT-UHFFFAOYSA-N 0.000 description 1
- AYRXSINWFIIFAE-UHFFFAOYSA-N O6-alpha-D-Galactopyranosyl-D-galactose Natural products OCC1OC(OCC(O)C(O)C(O)C(O)C=O)C(O)C(O)C1O AYRXSINWFIIFAE-UHFFFAOYSA-N 0.000 description 1
- 230000010718 Oxidation Activity Effects 0.000 description 1
- 229930182555 Penicillin Natural products 0.000 description 1
- JGSARLDLIJGVTE-MBNYWOFBSA-N Penicillin G Chemical compound N([C@H]1[C@H]2SC([C@@H](N2C1=O)C(O)=O)(C)C)C(=O)CC1=CC=CC=C1 JGSARLDLIJGVTE-MBNYWOFBSA-N 0.000 description 1
- 239000001888 Peptone Substances 0.000 description 1
- 108010080698 Peptones Proteins 0.000 description 1
- NGFMICBWJRZIBI-JZRPKSSGSA-N Salicin Natural products O([C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O1)c1c(CO)cccc1 NGFMICBWJRZIBI-JZRPKSSGSA-N 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 102000019197 Superoxide Dismutase Human genes 0.000 description 1
- 108010012715 Superoxide dismutase Proteins 0.000 description 1
- 239000004098 Tetracycline Substances 0.000 description 1
- HDTRYLNUVZCQOY-WSWWMNSNSA-N Trehalose Natural products O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 HDTRYLNUVZCQOY-WSWWMNSNSA-N 0.000 description 1
- 239000002253 acid Substances 0.000 description 1
- 230000004913 activation Effects 0.000 description 1
- 230000003044 adaptive effect Effects 0.000 description 1
- 230000006838 adverse reaction Effects 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- WQZGKKKJIJFFOK-DVKNGEFBSA-N alpha-D-glucose Chemical compound OC[C@H]1O[C@H](O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-DVKNGEFBSA-N 0.000 description 1
- NGFMICBWJRZIBI-UHFFFAOYSA-N alpha-salicin Natural products OC1C(O)C(O)C(CO)OC1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UHFFFAOYSA-N 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
- 230000003064 anti-oxidating effect Effects 0.000 description 1
- 238000009635 antibiotic susceptibility testing Methods 0.000 description 1
- PYMYPHUHKUWMLA-UHFFFAOYSA-N arabinose Natural products OCC(O)C(O)C(O)C=O PYMYPHUHKUWMLA-UHFFFAOYSA-N 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 235000015278 beef Nutrition 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- DRTQHJPVMGBUCF-PSQAKQOGSA-N beta-L-uridine Natural products O[C@H]1[C@@H](O)[C@H](CO)O[C@@H]1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-PSQAKQOGSA-N 0.000 description 1
- GUBGYTABKSRVRQ-QUYVBRFLSA-N beta-maltose Chemical compound OC[C@H]1O[C@H](O[C@H]2[C@H](O)[C@@H](O)[C@H](O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@@H]1O GUBGYTABKSRVRQ-QUYVBRFLSA-N 0.000 description 1
- 229940093761 bile salts Drugs 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 238000002815 broth microdilution Methods 0.000 description 1
- 229910000019 calcium carbonate Inorganic materials 0.000 description 1
- 229940041514 candida albicans extract Drugs 0.000 description 1
- 239000006143 cell culture medium Substances 0.000 description 1
- 229960002626 clarithromycin Drugs 0.000 description 1
- AGOYDEPGAOXOCK-KCBOHYOISA-N clarithromycin Chemical compound O([C@@H]1[C@@H](C)C(=O)O[C@@H]([C@@]([C@H](O)[C@@H](C)C(=O)[C@H](C)C[C@](C)([C@H](O[C@H]2[C@@H]([C@H](C[C@@H](C)O2)N(C)C)O)[C@H]1C)OC)(C)O)CC)[C@H]1C[C@@](C)(OC)[C@@H](O)[C@H](C)O1 AGOYDEPGAOXOCK-KCBOHYOISA-N 0.000 description 1
- 150000001875 compounds Chemical class 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- YXVFQADLFFNVDS-UHFFFAOYSA-N diammonium citrate Chemical compound [NH4+].[NH4+].[O-]C(=O)CC(O)(C(=O)O)CC([O-])=O YXVFQADLFFNVDS-UHFFFAOYSA-N 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 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 1
- 201000010099 disease Diseases 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000003028 enzyme activity measurement method Methods 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 235000021107 fermented food Nutrition 0.000 description 1
- MHMNJMPURVTYEJ-UHFFFAOYSA-N fluorescein-5-isothiocyanate Chemical compound O1C(=O)C2=CC(N=C=S)=CC=C2C21C1=CC=C(O)C=C1OC1=CC(O)=CC=C21 MHMNJMPURVTYEJ-UHFFFAOYSA-N 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000004927 fusion Effects 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 229960002518 gentamicin Drugs 0.000 description 1
- DLRVVLDZNNYCBX-CQUJWQHSSA-N gentiobiose Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC[C@@H]1[C@@H](O)[C@H](O)[C@@H](O)C(O)O1 DLRVVLDZNNYCBX-CQUJWQHSSA-N 0.000 description 1
- 229960002989 glutamic acid Drugs 0.000 description 1
- 230000002641 glycemic effect Effects 0.000 description 1
- 125000002887 hydroxy group Chemical group [H]O* 0.000 description 1
- 229940126904 hypoglycaemic agent Drugs 0.000 description 1
- 230000000415 inactivating effect Effects 0.000 description 1
- 238000011081 inoculation Methods 0.000 description 1
- 229940125396 insulin Drugs 0.000 description 1
- 230000010354 integration Effects 0.000 description 1
- 230000003834 intracellular effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- WRUGWIBCXHJTDG-UHFFFAOYSA-L magnesium sulfate heptahydrate Chemical compound O.O.O.O.O.O.O.[Mg+2].[O-]S([O-])(=O)=O WRUGWIBCXHJTDG-UHFFFAOYSA-L 0.000 description 1
- 229940061634 magnesium sulfate heptahydrate Drugs 0.000 description 1
- SCVOEYLBXCPATR-UHFFFAOYSA-L manganese(II) sulfate pentahydrate Chemical compound O.O.O.O.O.[Mn+2].[O-]S([O-])(=O)=O SCVOEYLBXCPATR-UHFFFAOYSA-L 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 230000004060 metabolic process Effects 0.000 description 1
- CWKLZLBVOJRSOM-UHFFFAOYSA-N methyl pyruvate Chemical compound COC(=O)C(C)=O CWKLZLBVOJRSOM-UHFFFAOYSA-N 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 229910000402 monopotassium phosphate Inorganic materials 0.000 description 1
- 235000019796 monopotassium phosphate Nutrition 0.000 description 1
- 229940049954 penicillin Drugs 0.000 description 1
- 235000019319 peptone Nutrition 0.000 description 1
- 230000035790 physiological processes and functions Effects 0.000 description 1
- 235000021110 pickles Nutrition 0.000 description 1
- 229920000136 polysorbate Polymers 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000029537 positive regulation of insulin secretion Effects 0.000 description 1
- GNSKLFRGEWLPPA-UHFFFAOYSA-M potassium dihydrogen phosphate Chemical compound [K+].OP(O)([O-])=O GNSKLFRGEWLPPA-UHFFFAOYSA-M 0.000 description 1
- LWIHDJKSTIGBAC-UHFFFAOYSA-K potassium phosphate Substances [K+].[K+].[K+].[O-]P([O-])([O-])=O LWIHDJKSTIGBAC-UHFFFAOYSA-K 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 238000012847 principal component analysis method Methods 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- NGFMICBWJRZIBI-UJPOAAIJSA-N salicin Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC=C1CO NGFMICBWJRZIBI-UJPOAAIJSA-N 0.000 description 1
- 229940120668 salicin Drugs 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 235000020183 skimmed milk Nutrition 0.000 description 1
- 239000001632 sodium acetate Substances 0.000 description 1
- 235000017281 sodium acetate Nutrition 0.000 description 1
- 239000000600 sorbitol Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 230000003068 static effect Effects 0.000 description 1
- 239000011550 stock solution Substances 0.000 description 1
- 229960005322 streptomycin Drugs 0.000 description 1
- 150000008163 sugars Chemical class 0.000 description 1
- 239000013589 supplement Substances 0.000 description 1
- 229960002180 tetracycline Drugs 0.000 description 1
- 229930101283 tetracycline Natural products 0.000 description 1
- 235000019364 tetracycline Nutrition 0.000 description 1
- 150000003522 tetracyclines Chemical class 0.000 description 1
- DRTQHJPVMGBUCF-UHFFFAOYSA-N uracil arabinoside Natural products OC1C(O)C(CO)OC1N1C(=O)NC(=O)C=C1 DRTQHJPVMGBUCF-UHFFFAOYSA-N 0.000 description 1
- 229940045145 uridine Drugs 0.000 description 1
- 235000013311 vegetables Nutrition 0.000 description 1
- 239000012138 yeast extract Substances 0.000 description 1
- 235000013618 yogurt Nutrition 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
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- 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
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P39/00—General protective or antinoxious agents
- A61P39/06—Free radical scavengers or antioxidants
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/169—Plantarum
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12R—INDEXING SCHEME ASSOCIATED WITH SUBCLASSES C12C - C12Q, RELATING TO MICROORGANISMS
- C12R2001/00—Microorganisms ; Processes using microorganisms
- C12R2001/01—Bacteria or Actinomycetales ; using bacteria or Actinomycetales
- C12R2001/225—Lactobacillus
- C12R2001/25—Lactobacillus plantarum
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Biotechnology (AREA)
- Zoology (AREA)
- Genetics & Genomics (AREA)
- Microbiology (AREA)
- Wood Science & Technology (AREA)
- Mycology (AREA)
- Pharmacology & Pharmacy (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Biochemistry (AREA)
- Veterinary Medicine (AREA)
- Diabetes (AREA)
- Virology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Tropical Medicine & Parasitology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Biomedical Technology (AREA)
- General Engineering & Computer Science (AREA)
- Food Science & Technology (AREA)
- Epidemiology (AREA)
- Emergency Medicine (AREA)
- Endocrinology (AREA)
- Hematology (AREA)
- Obesity (AREA)
- Molecular Biology (AREA)
- Polymers & Plastics (AREA)
- Nutrition Science (AREA)
- Toxicology (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Measuring Or Testing Involving Enzymes Or Micro-Organisms (AREA)
Abstract
The invention provides Lactobacillus plantarum, the preservation number of which is CCTCC M2019591, the Latin name is Lactobacillus plantarum CT 152; provides a medicament with the function of reducing blood sugar, which adopts the lactobacillus plantarum. And provides a method for screening lactobacillus plantarum, which comprises the following steps: s1: carrying out probiotic index determination; s2: measuring the oxidation resistance; s3: detecting the anti-diabetic properties of the lactic acid bacteria; s4: in vivo experiments were performed. The invention systematically establishes an in-vitro efficient screening method of the blood sugar-reducing lactic acid bacteria, obtains the invention bacterial strain CCTCC M2019591 with better blood sugar-reducing effect by screening in a mode of combining PCA and AHP, and obtains the effect verification of in-vivo experiments.
Description
Technical Field
The invention particularly relates to lactobacillus plantarum with a blood sugar reducing effect, and a screening method and an application medicament thereof.
Background
The international diabetes consortium published an eighth version of the global diabetes map. The results show that worldwide adult patients with diabetes (20-79 years) have reached 4.25 billion from 1.51 billion in 2000 to 2017, an increase of nearly 2-fold. It is expected that by 2045 years, diabetics may reach 6.29 billion. While the Chinese diabetes patients are the first in the world, diabetes is recognized as the third most fatal disease in the world, the incidence rate of the diabetes is increased year by year, and the proportion of the type II diabetes patients is up to more than 90%. At present, the purpose of treating type II diabetes is achieved mainly by lowering blood sugar through medicines. Hypoglycemic drugs for diabetes mellitus generally cause a series of adverse reactions, such as diarrhea, drug resistance, secondary failure and the like. In recent years, due to the nature and safety of lactic acid bacteria, it has become a focus of research on the prevention and treatment of diabetes.
Lactic Acid Bacteria (LAB) refer to a general term for a class of non-spore, gram-positive bacteria that ferment sugars, the main product being Lactic acid. Research in recent years shows that lactic acid bacteria have various physiological functions as beneficial bacteria groups which are settled in intestinal tracts; maintain intestinal microecological balance, influence blood sugar metabolism, etc. At present, an in vitro screening method aiming at a comprehensive system of the blood sugar reducing lactic acid bacteria is not established, and in vivo animal experiment screening is limited by factors in time and price, so that evaluation and integration of the existing in vitro screening method of the blood sugar reducing lactic acid bacteria are urgently needed, a set of screening method for the blood sugar reducing lactic acid bacteria comprehensively and systematically is established, and a certain technical and method support is provided for the mechanism research of the blood sugar reducing lactic acid bacteria. Researchers such as the Chenpei and the like screen 2 strains of lactic acid bacteria for reducing blood sugar from the outside of the body by using a principal component analysis method, and verify the blood sugar reducing effect in the body, but the screening indexes are not comprehensive enough, and the safety of the lactic acid bacteria is not evaluated. The invention integrates and supplements according to the screening indexes of in vitro functional bacteria of earlier researchers, and establishes the screening method of the high-efficiency blood sugar reducing lactic acid bacteria by combining a Principal Component Analysis (PCA) method and an Analytic Hierarchy Process (AHP) method, and is proved in vivo, thereby providing a certain technical and method support for the research of the blood sugar reducing mechanism of the lactic acid bacteria.
Disclosure of Invention
One of the technical problems solved by the invention is the establishment of a screening method of in vitro hypoglycemic lactic acid bacteria; the second technical problem to be solved by the invention is to provide a strain of Lactobacillus plantarum CT152 which is obtained by applying the screening method and has higher survival rate in an in vitro simulated gastrointestinal fluid and bile salt environment through in vivo verification; and has certain anti-oxidation and anti-diabetic properties; the second technical problem to be solved by the invention is to provide a hypoglycemic agent applied to the hypoglycemic strain Lactobacillus plantarum CT 152.
In order to meet the requirements, the technical scheme adopted by the invention is as follows: m2019591, Latin with the name of Lactobacillus plantarum CT152, a preservation place of China center for type culture Collection, wherein the preservation place is in the eight-channel 229 # Wuhan university school in Wuchang district, Wuhan City, Hubei province, and the preservation date is 7 months and 30 days in 2019; providing a medicament with the function of reducing blood sugar, which adopts the lactobacillus plantarum; provides a screening method of lactobacillus plantarum, which comprises the following steps:
s1: performing probiotic index determination, including survival rate determination in simulated gastrointestinal fluid, bile salt tolerance determination, adhesion capability determination to Caco-2 cells, and antibiotic resistance detection;
s2: performing antioxidant capacity measurement, including DPPH removing capacity measurement, hydroxyl radical removing capacity measurement, lipid peroxidation resisting capacity measurement, reducing capacity measurement, and enzyme activity measurement of GSH-Px and T-SOD of lactobacillus;
s3 testing the antidiabetic properties of lactic acid bacteria, including the determination of the rate of inhibition of the activity of α -glucosidase in rat intestinal tract and the experiment of intervention of lactic acid bacteria in insulin secretion from high-sugar stimulated islet cells, and
s4: and (3) carrying out in-vivo experiment verification, including the construction of a type 2 diabetes model rat and the intragastric administration treatment.
The invention provides a high-efficiency screening method of in-vitro hypoglycemic lactobacillus, which can provide technical and method support for subsequent screening of functional lactobacillus to a certain extent, and a potential hypoglycemic strain CCTCC M2019591 is obtained by screening through the method, in the aspect of probiotics, the gastrointestinal fluid tolerance of the inventive strain is good, the survival rate reaches 97.62%, the tolerance and the adhesion capacity to Caco-2 cells are stronger than those of a positive control strain, the adhesion rate reaches 18.67%, the bacterial strain has high oxidation resistance, the cell-free extract and the intact cells reach about 15% -16% in DPPH clearance capacity, in the aspect of hydroxyl radical scavenging capacity, the inventive strain has equivalent capacity to a commercial control strain, in the aspect of lipid peroxidation resistance, the fermented supernatant and the intact cells of the inventive strain are stronger, respectively reach 8.48% and 15.64%, in the aspects of GSH-Px and T-SOD enzyme activity, the bacterial strain CCTCC M2019591 is also stronger, in the aspect of α -glucosidase extracted from rat intestinal tracts, the strain can reach about 15% and the safety inhibition of insulin resistance of the inventive strain can be proved by the comprehensive test of the insulin resistance of the inventive strain, and the insulin resistance of the inventive strain can be improved by the high insulin resistance of the experimental strain, and the research of the insulin resistance of the inventive strain can be proved by the invention, and the research of the invention, and the animal insulin resistance of the invention can be improved by the research of the invention.
Drawings
The accompanying drawings, which are included to provide a further understanding of the application and are incorporated in and constitute a part of this application, illustrate embodiment(s) of the application and together with the description serve to explain the application and not to limit the application. In the drawings:
FIG. 1 schematically shows a schematic diagram of the effect of Lactobacillus plantarum on the insulin secretion capacity of INS-1 cells according to one embodiment of the present application.
FIG. 2 schematically shows a graph of the adhesion rate of Lactobacillus plantarum to Caco-2 cells, according to one embodiment of the present application.
Fig. 3 schematically shows a graph of random blood glucose change values for rats around intragastric gavage according to one embodiment of the present application.
FIG. 4 schematically shows the random blood glucose variability of rats gavaged for six weeks according to one embodiment of the present application.
Detailed Description
In order to make the objects, technical solutions and advantages of the present application more apparent, the present application will be described in further detail with reference to the accompanying drawings and specific embodiments.
In the following description, references to "one embodiment," "an embodiment," "one example," "an example," etc., indicate that the embodiment or example so described may include a particular feature, structure, characteristic, property, element, or limitation, but every embodiment or example does not necessarily include the particular feature, structure, characteristic, property, element, or limitation. Moreover, repeated use of the phrase "in accordance with an embodiment of the present application" although it may possibly refer to the same embodiment, does not necessarily refer to the same embodiment.
Certain features that are well known to those skilled in the art have been omitted from the following description for the sake of simplicity.
According to one embodiment of the application, the Lactobacillus plantarum is provided, and the preservation number of the Lactobacillus plantarum is CCTCC M2019591, and the Latin name is Lactobacillus plantarum CT 152; and provides a medicament having hypoglycemic activity, which comprises the Lactobacillus plantarum described in claim 1.
According to an embodiment of the present application, a method for screening lactobacillus plantarum is provided, which specifically includes the following steps:
(1) and (3) probiotic index determination:
1) simulated survival in gastrointestinal fluids: after the lactic acid bacteria were resuspended in 0.85% physiological saline, the density of the bacterial fluid was adjusted to 1X 109CFU/mL in simulated gastric fluid (pH 3.0). After mixing uniformly, placing the mixture at 37 ℃ for culturing for 1, 2 and 3 hours, and then respectively counting the number of viable bacteria. After culturing in simulated gastric fluid for 3h, 1mL of the culture medium was added to 9mL of simulated intestinal fluid (pH 8.0), mixed and cultured at 37 ℃. Viable count was measured at 2, 4 and 8h respectively. The survival rate of the cells is calculated by culturing the cells for 48 hours at 37 ℃ in an MRS solid medium.
Survival (%) [ log CFU N1/log CFU N ] x 100%;
n1-the viable count of lactic acid bacteria after simulated gastrointestinal fluid treatment, and N0-the viable count of lactic acid bacteria before non-treatment.
2) Tolerance of bile salt: inoculating lactobacillus cultured for 18h 2% -5% in MRS-THIO (MRS contains 0.2% sodium thioglycolate) culture medium with or without 0.3% bile salt. Culturing the mixture at 37 deg.C for 16-24 hr at 600nm at intervals; and (3) measuring the light absorption value, and recording the time required by the light absorption value of the culture medium without the bile salt and the light absorption value of the culture medium with the bile salt to reach 0.3 unit, wherein the two time differences are the delay time (LT) of the growth of the lactic acid bacteria in the bile salt, and the shorter the LT is, the stronger the ability of the lactic acid bacteria to tolerate the bile salt is.
3) Adhesion capability to Caco-2 cells: caco-2 cells were cultured in high-sugar DMEM medium containing 20% fetal bovine serum, 1% L-glutamine, 1% nonessential amino acids and 1% diabase at 37 ℃ in a 5% CO2 incubator, and the medium was changed every other day until the cell proliferation fusion rate reached about 80%, and then 0.25% pancreatin-digested cells containing 0.02% EDTA were passaged at 1:2-1: 3. Cells in the logarithmic growth phase were taken for the experiments.
Freshly cultured lactic acid bacteria were washed twice with PBS and the bacterial concentration was adjusted to 1X 109CFU/mL, suspended in 100mg/mL Fluorescein Isothiocyanate (FIFC), exposed to 37 ℃ in the dark for 0.5-1h, and washed 4 times with PBS to remove unbound FIFC. Caco-2 cells were cultured at 4X 105-5×105cells/well were inoculated in 6-well plates and cultured at 37 ℃ for 16-24 h. FIFC-labeled lactic acid bacteria were resuspended in DMEM medium and adjusted to a density of 1X 109 CFU/mL. Subjecting the strain to fermentationAdding the suspension into Caco-2 cell monolayer, acting at 37 deg.C for 1-2h, washing with sterile PBS for 3 times, removing non-adhered bacteria, adding pancreatin into each cultured well, and adding 0.4mL complete culture medium to stop reaction when cells completely drop. Collecting liquid, and measuring the fluorescence intensity by using a multifunctional microplate reader to calculate the adhesion rate of bacteria. A and A0 represent the fluorescence intensity before and after adhesion of lactic acid bacteria to cells, respectively.
The adhesion rate (%) ═ Log a/Log a0 × 100%.
4) And (3) detecting antibiotic resistance:
the strains were analyzed for antibiotic resistance according to the European Committee for drug sensitivity of microorganisms (http:// www.eucast.org) with respect to the sensitivity threshold X of the microorganisms to antibiotics (http:// www.eucast.org/mic _ distributions /). When the Minimum Inhibitory Concentration (MIC) of the isolated strain is less than or equal to X [ mu ] g/mL, the isolated strain is a sensitive strain; otherwise, the strain is a drug-resistant strain.
MIC determination was performed by broth dilution. Commonly used antibiotics are prepared into corresponding stock solutions respectively, and MRS liquid culture medium is diluted into using solution in a 2-fold gradient manner. After 198. mu.L of MRS liquid medium containing antibiotics at different concentrations was added to a sterile 96-well plate, 2. mu.L of a culture solution of the strain (about 1.0X 107CFU/mL) was inoculated, and after static culture at 37 ℃ for 24 hours, MICs of the different strains were counted. Each group was repeated 3 times, and a control group was set.
(2) And (3) measuring the antioxidant capacity:
1) determination of DPPH scavenging ability of lactic acid bacteria: adding 1mL of lactobacillus suspension, cell-free extract or fermentation supernatant into the reaction system, adding 1mL of 0.2mmol/L of absolute ethanol of DPPH, shaking, mixing well, reacting at room temperature in a dark place for 30min, centrifuging at 6000rpm for 10min, taking supernatant, and measuring the absorbance (OD value) of the sample at the wavelength of 517 nm. The sample solution was replaced with an equal volume of saline as a control group, and the volume of a mixture of saline and absolute ethanol was set to zero as a blank. Calculated according to the following formula:
clearance of DPPH free radical [1-A ]517(sample)/A517(control)]×100％
2) And (3) determining the hydroxyl radical scavenging capacity of lactic acid bacteria: the hydroxyl radical kit determination of Nanjing established biological company is not strictly carried out according to the instruction.
3) And (3) measuring the lipid peroxidation resisting capacity of the lactic acid bacteria: adding 1mL of linoleic acid emulsion into 0.5mL of PBS with the pH value of 7.4, adding 0.2mL of 0.01% ferrous sulfate and 0.01% ascorbic acid, then adding 0.2m-0.5mL of fermentation supernatant, bacterial suspension or cell extract, uniformly mixing, placing in a water bath at 37 ℃ for reaction for 12h, adding 0.2mL of 0.4% trichloroacetic acid, 2mL of 0.8% thiobarbituric acid and 0.2mL of 0.4% BHT into 2mL of reaction solution, shaking, fully mixing, reacting at 100 ℃ for 30min, adding 2mL of trichloromethane for extraction after a sample is cooled, centrifuging for 10min, and collecting supernatant. OD values of the samples were measured at 532nm wavelength with PBS as a blank. The lipid peroxidation resistance of lactic acid bacteria was calculated according to the following formula: anti-lipid peroxidation rate ═ 1-A532(sample)/A532(blank)]×100％。
4) Determination of the reducing power of lactic acid bacteria: 0.5mL of the bacterial suspension or cell extract was added with 0.5mL of 1% potassium ferricyanide and 0.5mL of PBS (pH 6.6), shaken, mixed well, and bathed in water at 50 ℃ for 20min, and the sample was replaced with distilled water to serve as a control. After rapidly cooling in an ice bath, 0.5mL of 10% trichloroacetic acid was added, centrifugation was carried out at 3000rpm for 10min, 1mL of the supernatant was taken, 1mL of 0.1% Fe CL3 was added, and the OD value of the sample was measured at a wavelength of 700nm after 10min of reaction. The result uses cysteine as a standard to characterize the reducing power, and the calculation formula is as follows: reducing power [ A ]700(sample) -A700(control)/A700(control)]× 100％。
5) And (3) measuring the enzyme activity of GSH-Px and T-SOD of the lactic acid bacteria:
the enzyme activities of GSH-Px and T-SOD in lactobacillus fermentation supernatant and cell extract are detected by using Nanjing-built kit, and the specific operation is strictly according to the kit instruction.
(3) Anti-diabetic properties of lactic acid bacteria
1) Determination of rat intestinal α -glucosidase activity inhibition rate
To 150. mu.L of PBS (0.1M pH6.8) were added 75. mu.L of 20mM PNPG solution and 25. mu.L of the lactic acid bacteria to be testedFermenting the supernatant, cell-free extract and intact cells, adding the mixture into water bath at 37 deg.C for 10min, adding 50 μ L α -glucopsidase solution (0.17U/mL) extracted from rat intestinal tract, continuing reaction for 10min, adding 1mL0.1M Na2CO3As a reaction termination solution. Measuring the absorbance of the reaction solution at 405nm, the absorbance and p-nitrophenol (PNP)
) The reaction system adopts 0.1M PBS with pH of 6.8 as α -glucopsidase solution and blank control of the sample to be detected, and the inhibitory activity of the sample is calculated by the following formula.
Enzyme inhibition (%) [1- (C-D)/(A-B) ]. times.100%
Wherein A is the measured absorbance containing α -glucopsidase solution but no sample
B is the measured absorbance value of the solution without α -glucopsidase and the sample to be measured
C is the determination light absorption value of α -glucopsidase solution and sample to be measured
D is the measured absorbance value of the sample not containing α -glucopsidase solution but containing the sample to be measured
2) Experiment for intervention of lactobacillus in high-sugar stimulation of insulin secretion of islet cells
The lactobacillus is subjected to anaerobic passage for 2 times in MRS liquid culture medium, and is incubated for 18 hours at 37 ℃ each time, so that the strain activity reaches the highest value. The bacterial suspension is subjected to gradient dilution by a sterilized PBS buffer solution, a plate pouring method is adopted, the bacterial suspension is cultured for 24-48h at 37 ℃ on an MRS solid culture medium, and the number of viable bacteria is determined (ensuring that each plate has 30-300 viable bacteria colonies). According to the number of viable bacteria of the bacterial powder, 1 × 10 of the bacterial powder is prepared9And (3) carrying out 120kw ultrasonic crushing on the bacterial suspension of CFU/mL, then inactivating at 65 ℃ for 30-60min, and centrifuging at 8000 Xg to obtain a thallus mixture precipitate. Resuspend with INS-1 cell culture medium without double antibody, use as culture medium of GSIS experiment stimulating cells, store at 4 deg.C for use.
INS-1 cells at 4X 105-5×105Inoculating cells/mL into a 6-well plate, incubating overnight for cell adherence, and then incubating for 16-24h by using a cell culture solution containing an inactivated bacterial solution to perform intervention action of different probiotic strains on the cells. After the treatment of each group of cells was completed, the cells were washed with Phosphate Buffered Saline (PBS)2 times, adding high-sugar INS-1 complete culture solution with glucose concentration of 16.7mmol/L, and incubating at 37 deg.C for 0.5-1 h. Cell supernatants were then collected and the amount of insulin secretion from each group under high glucose challenge was measured by ELISA. ELISA detection steps are carried out according to the kit instructions, and the absorbance is detected by an enzyme-linked immunosorbent assay (at 450 nm). The OD value is used as the ordinate (Y) and the standard substance concentration is used as the abscissa (X), a standard curve is drawn, and the concentrations of other samples are calculated according to the standard curve.
(4) Verification of in vivo experiments
1) Construction of type 2 diabetes model rat
All experimental animals were fed via the western animal house of Sichuan university. The experimental animal room was kept at an appropriate temperature and humidity throughout the year and strictly followed the 12h day and 12h night cycle standard. 70 male Wistar rats of 6-8 weeks of age were randomized into low-fat and high-fat groups after 5 days of adaptive basal feeding. For the first four weeks, each group was fed high fat diet except the low fat group, and the weight measurements of the rats were recorded at fixed times per week.
In the fifth week, all rats were fasted for 12-18h and, except for the low fat group, the rats in each group were injected with 30-40mg/kg body weight of freshly prepared streptozotocin STZ. An amount of STZ was dissolved in citrate-sodium citrate buffer (pH 4.5) and ready to use, stored in an ice bath, and injected into the low fat group in citrate-sodium citrate buffer (pH 4.5). Ensuring sufficient sterile water and high-fat diet in 72h after molding, changing padding every day, and taking diabetic rats with random blood sugar higher than 16.7mmol/L or fasting blood sugar higher than 11.7mmol/L after 72 h.
2) Gavage treatment
Dividing the rat into model group, LGG group, and CT152 group, wherein the model group uses 0.85% physiological saline as control, and LGG and CCTCC M2019591 are 1 × 109CFU/day was treated for a 6-8 week period with gavage.
In-vivo experimental verification the random blood glucose change values of rats at 6 weeks of gastric lavage are shown in figure 4.
Preparing a lactic acid bacteria agent: 1) inoculating CCTCC M2019591 strain on a solid slant culture medium, culturing at 37 ℃ for 16-24h for activation, then inoculating the cultured solid slant culture medium into a seed liquid culture medium under an aseptic condition, and culturing at 37 ℃ for 16-24h to prepare a primary seed liquid; inoculating the first-stage seed liquid into a seed liquid culture medium according to the inoculation amount of 2-5%, standing and culturing at 37 ℃ for 16-24h, and collecting the fermentation liquid. 2) And (3) immediately centrifuging the fermentation liquor after fermentation is finished, washing the fermentation liquor by using clear water, repeating the steps for 2-3 times, and then performing fermentation according to the following steps: the protective agent is 1:1-1:2 (the protective agent can be trehalose, skim milk powder, sorbitol, mannitol, etc. or a compound protective agent taking the protective agent as raw materials), and the lactic acid bacteria agent is obtained after freeze drying and crushing.
The invention provides a screening method of Lactobacillus plantarum capable of reducing blood sugar widely and systematically, and a Lactobacillus plantarum strain with probiotics, high oxidation activity and potential blood sugar reducing efficacy is obtained by screening with the method, the strain separating screen is selected from traditional fermented food pickle, is named as Lactobacillus plantarum CT152, has a preservation number of CCTCC M2019591, has a preservation date of 2019, 7 and 30 days, and is preserved in China center for type culture preservation in Wuhan City.
The strain is round on MRS culture medium, has smooth and compact surface, white color, and occasionally has light yellow or dark yellow color, and uniform colony size, and has the physiological characteristics of utilizing N-acetyl-D galactosamine, gentiobiose, α -D-glucose, D-cellobiose, D-fructose, mannose, mannitol, maltose, β -methyl-D-glucose, D-ketorolose, D-ribose, salicin, D-trehalose, D-xylose, methyl pyruvate, glycerol, uridine, no starch, mannan, D-galactose, L-trehalose, L-glutamic acid, etc.
The method comprises the steps of separating and purifying 120 lactic acid bacteria from pickled vegetables and fermented yoghourt, obtaining 18 candidate experimental strains through acid-resistant and cholate-resistant primary screening, comprehensively evaluating the hypoglycemic capability of the candidate 20 strains through detecting probiotic indexes (simulating the tolerance capability of gastrointestinal fluids, the cholate tolerance capability and the adhesion capability to Caco-2 cells), measuring antioxidant indexes (the scavenging capability to DPPH free radicals, the scavenging capability to hydroxyl free radicals, the lipid peroxidation resistance and the reduction capability, and the activity measurement to glutathione peroxidase GSH-Px and total superoxide dismutase T-SOD), measuring an antidiabetic index (α -glucosidase inhibition rate and the insulin production capability of rat islet cell INS-1 under the intervention of lactic acid bacteria), obtaining the probiotic, antioxidant and antidiabetic index scores through a Principal Component Analysis (PCA), obtaining a hypoglycemic comprehensive index by using an analytic hierarchy process (score P), and obtaining a hypoglycemic comprehensive index by using a commercial LGG (Lactobacillus strain GG 53103) as a contrast, wherein the hypoglycemic comprehensive strain is selected as a higher than the AHG.
The culture medium formula (1L) adopted by the invention is as follows: 10g of peptone, 20g of glucose, 5g of yeast extract, 10g of beef extract, 2g of monopotassium phosphate, 2g of ammonium hydrogen citrate, 5g of sodium acetate, 0.5g of magnesium sulfate heptahydrate, 0.25g of manganese sulfate pentahydrate, 801mL of tween, 20g of agar (solid culture) and 7.5g of calcium carbonate (solid separation culture).
The test records of this example are shown in tables 1-8 below, wherein:
table 1 is a measure of the resistance of lactic acid bacteria to intestinal gastric juice;
table 2 shows the determination of the tolerance of lactic acid bacteria to bile salts;
table 3 lactic acid bacteria DPPH scavenging and hydroxyl radical scavenging capacity;
table 4 lactic acid bacteria anti-lipid peroxidation and reduction capacity assay;
TABLE 5 lactic acid bacteria fermentation supernatants and viability of intracellular GSH-Px and T-SOD;
TABLE 6 inhibition rate of lactic acid bacteria on α -glucopsidase activity extracted from rat intestine;
table 7 lactic acid bacteria antibiotic susceptibility test results;
table 8 combined glycemic index score by PCA and AHP methods;
table 1:
table 2:
table 3:
table 4:
table 5:
table 6:
table 7:
GEN, gentamicin; AMP, penicillin; KM, clarithromycin; STR, streptomycin; TET, tetracycline;
ERY, erythromycin; CLI, clindamycin; VAN, vancomycin; RIF, rifampin; CM, chloramphenicol.
Table 8:
the above-described embodiments are merely illustrative of several embodiments of the present invention, which are described in more detail and detail, but are not to be construed as limiting the scope of the present invention. It should be noted that, for a person skilled in the art, several variations and modifications can be made without departing from the inventive concept, which falls within the scope of the present invention. Therefore, the protection scope of the present invention should be subject to the claims.
Claims (12)
1. A lactobacillus plantarum strain characterized by: the Lactobacillus plantarum has a preservation number of CCTCC M2019591 and a Latin name of Lactobacillus plantarum CT 152.
2. A medicine and food with the function of reducing blood sugar are characterized in that: the use of the Lactobacillus plantarum strain defined in claim 1.
3. A method for screening Lactobacillus plantarum is characterized by comprising the following steps:
s1: performing probiotic index determination, including survival rate determination in simulated gastrointestinal fluid, bile salt tolerance determination, adhesion capability determination to Caco-2 cells, and antibiotic resistance detection;
s2: measuring the oxidation resistance;
s3, performing detection on the anti-diabetic characteristics of the lactic acid bacteria, including determination of the activity inhibition rate of α -glucosidase in rat intestinal tracts and experiment of intervention of the lactic acid bacteria in insulin secretion of high-sugar-stimulated islet cells.
4. The method for screening Lactobacillus plantarum according to claim 3, wherein: the step of measuring the antioxidant capacity specifically comprises the following steps: the method comprises the steps of measuring DPPH removing capacity of the lactobacillus, measuring hydroxyl radical removing capacity of the lactobacillus, measuring lipid peroxidation resisting capacity of the lactobacillus, measuring reducing capacity of the lactobacillus and measuring enzyme activity of GSH-Px and T-SOD of the lactobacillus.
5. The method for screening Lactobacillus plantarum according to claim 3, wherein: the method also comprises the step of carrying out in vivo experimental verification, and specifically comprises the steps of constructing a type 2 diabetes model rat and carrying out intragastric administration treatment.
6. The method for screening Lactobacillus plantarum according to claim 3, wherein: the specific steps of the determination of viability in simulated gastrointestinal fluids comprise: after resuspending the lactic acid bacteria in 0.85% physiological saline,adjusting the density of the bacterial liquid to 1 × 10 in simulated gastric fluid with pH of 39CFU/mL, mixing, placing in 37 deg.C, culturing for 1, 2 and 3h, respectively counting viable count, culturing in simulated gastric fluid for 3h, adding 1mL culture solution into 9mL simulated intestinal fluid with pH of 8, mixing, culturing at 37 deg.C, detecting viable count for 2, 4 and 8h, culturing at 37 deg.C with MRS solid culture medium for 24-48h, calculating survival rate,
survival (%) [ log CFU N1/log CFU N ] × 100%
N1The viable count of lactobacillus after simulated gastrointestinal fluid treatment, N0Live count of untreated lactic acid bacteria.
7. The method for screening Lactobacillus plantarum according to claim 3, wherein: the steps of the bile salt tolerance determination comprise: inoculating 2-5% of lactobacillus cultured for 18-24 h into MRS-THIO culture medium containing or not containing 0.3% bile salt, respectively, and culturing the mixture at 37 deg.C for 16-24h at intervals of 600 nm; measuring a light absorption value, and recording the time required by the light absorption value of the culture medium without the bile salt and with the bile salt when the light absorption value reaches 0.3 unit, wherein the two time differences are the delay time of the growth of the lactic acid bacteria in the bile salt, the delay time is LT, and the shorter the LT is, the lactic acid bacteria have stronger bile salt tolerance; MRS in the MRS-THIO culture medium contains 0.2 percent of sodium thioglycolate.
8. The method for screening Lactobacillus plantarum according to claim 3, wherein: the step of determining the adhesion capability to Caco-2 cells comprises: caco-2 cells were grown in high glucose DMEM medium containing 20% fetal bovine serum, 1% L-glutamine, 1% non-essential amino acids and 1% diabodies at 37 deg.C with 5% CO2Culturing in the incubator, replacing the culture solution every other day, digesting the cells with 0.25% of pancreatin containing 0.02% of EDTA when the cell proliferation and fusion rate reaches about 80%, carrying out passage according to 1:2-1:3, and taking the cells in logarithmic growth phase for test;
freshly cultured lactic acid bacteria were washed twice with PBS and the bacterial concentration was adjusted to 1X 109CFU/mL, suspended at 100mg/mTreating L-fluorescein isothiocyanate in dark at 37 deg.C for 0.5-1 hr, washing with PBS 4 times to remove unbound FIFC, and washing Caco-2 cells at 4 × 105-5×105Inoculating cell/well into 6-well plate, culturing at 37 deg.C for 16-24 hr, re-suspending the FIFC-labeled lactobacillus in DMEM medium, and adjusting the density to 1 × 109CFU/mL, adding the bacterial suspension into a Caco-2 cell monolayer, acting for 1-2h at 37 ℃, washing for 3 times by using sterile PBS, washing off unadhered bacteria, adding pancreatin into each cultured hole for acting, adding 0.4mL of complete culture medium to stop reaction after the cells completely fall off, collecting liquid, and measuring the fluorescence intensity by using a multifunctional microplate reader to calculate the adhesion rate of the bacteria;
adhesion ratio (%) ═ Log A/Log A0×100％；
Wherein A and A0 represent fluorescence intensities before and after adhesion of lactic acid bacteria to cells, respectively.
9. The method for screening Lactobacillus plantarum according to claim 3, wherein: the method for measuring the DPPH removing capacity of the lactic acid bacteria comprises the following steps: adding 1mL of lactobacillus suspension, a cell-free extract or a fermentation supernatant into a reaction system, adding 1mL0.2mmol/L of DPPH absolute ethyl alcohol, oscillating, fully mixing uniformly, reacting for 30min in a dark place at room temperature, centrifuging at 6000rpm for 10min, taking the supernatant, measuring the absorbance value (OD value) of a sample at the wavelength of 517nm, replacing the sample solution with physiological saline with the same volume as a control group, adjusting the blank to zero by using the mixed solution of the physiological saline and the absolute ethyl alcohol with the same volume as the control group, and calculating according to the following formula:
clearance of DPPH free radical [1-A ]517(sample)/A517(control)]×100％；
The step of measuring the lipid peroxidation resisting capacity of the lactic acid bacteria comprises the following steps: adding 1mL of linoleic acid emulsion into 0.5mL of PBS with the pH value of 7.4, adding 0.2mL of 0.01% ferrous sulfate and 0.01% ascorbic acid, then adding 0.5mL of fermentation supernatant, bacterial suspension or cell extract, uniformly mixing, placing in a water bath with the temperature of 37 ℃ for reaction for 12 hours, taking 2mL of reaction solution, adding 0.2mL of 0.4% trichloroacetic acid, 2mL of 0.8% thiobarbituric acid and 0.2mL of 0.4% BHT, shaking, fully mixing, reacting at the temperature of 100 ℃ for 30 minutes, adding 2mL of trichloromethane for extraction after a sample is cooled, centrifuging for 10 minutes, collecting the supernatant, measuring the OD value of the sample at the wavelength of 532nm, taking PBS as a blank control, and calculating the lipid peroxidation resistance of the lactic acid bacteria according to the following formula:
anti-lipid peroxidation rate ═ 1-A532(sample)/A532(blank)]×100％；
The step of measuring the reducing power of the lactic acid bacteria comprises the following steps: adding 0.5mL of 1% potassium ferricyanide and 0.5mL of PBS with 6.6% of 0.5mL of pH into 0.5mL of bacterial suspension or cell extract, shaking, mixing well, bathing in water at 50 ℃ for 20min, replacing the sample with distilled water to serve as a control group, rapidly cooling in ice bath, adding 0.5mL of 10% trichloroacetic acid, centrifuging at 3000rpm for 10min, collecting 1mL of supernatant, adding 1mL of FeCl with 0.1% of 1mL3And after the reaction is carried out for 10min, determining the OD value of the sample under the wavelength of 700nm, and using cysteine as a standard to characterize the reducing force according to the result, wherein the calculation formula is as follows:
reducing power [ A ]700(sample) -A700(control)/A700(control)]×100％。
10. The method for screening Lactobacillus plantarum according to claim 3, wherein the step of determining the inhibition rate of α -glucosidase activity in rat intestine comprises adding 75. mu.L of 20mM PNPG solution and 25. mu.L of PBS (0.1M pH6.8) to 150. mu.L of PBS (0.1M pH 6.8. PBS has a pH of 6.8. fermentation supernatant, cell-free extract and intact cells of the lactic acid bacteria to be tested, subjecting the mixture to water bath at 37 ℃ for 10min, adding 50. mu.L of α -glucosidase solution extracted from rat intestine, wherein α -glucosidase solution is 0.17U/mL, continuing the reaction for 10min, adding 1mL0.1M Na2CO3As reaction termination liquid, measuring the light absorption value of the reaction liquid at 405nm, wherein the light absorption value is in direct proportion to the free amount of the p-nitrophenol, 0.1M PBS with pH of 6.8 is adopted in the reaction system as α -glucopyranosase solution and blank contrast of a sample to be measured, calculating the inhibitory activity of the sample by adopting the following formula,
enzyme inhibition (%) [1- (C-D)/(a-B) ] × 100%;
wherein A is the measured absorbance containing α -glucopsidase solution but not containing sample;
b is a determination light absorption value of a solution without α -glucopsidase and a sample to be detected;
c is a determination light absorption value of a solution containing α -glucopsidase and a sample to be detected;
d is the measured absorbance of the solution without α -glucopsidase but with the sample to be measured.
11. The method for screening Lactobacillus plantarum according to claim 3, wherein: the step of the experiment that the lactobacillus intervenes the insulin secretion of the high-sugar stimulation islet cells comprises the following steps: anaerobic passage of lactobacillus in MRS liquid culture medium for 2 times, incubating at 37 deg.C for 18 hr to maximize the strain activity, diluting the bacterial suspension with sterilized PBS buffer solution, culturing at 37 deg.C for 24-48 hr by plate pouring method, determining viable bacteria number, ensuring viable bacteria colony of each plate 30-300, and preparing 1 × 10 viable bacteria number according to the bacteria powder9120kw ultrasonic crushing of CFU/mL bacterial suspension, 30-60min inactivation treatment at 65 ℃, 8000 Xg centrifugation to obtain thalli mixture precipitate, resuspending with INS-1 cell culture solution without double antibody, storing at 4 ℃ as culture solution of GSIS experiment stimulating cells for later use,
INS-1 cells at 4X 105-5×105Inoculating cells/mL into a 6-well plate at a density of cells, incubating overnight for cell adherence, then incubating for 16-24h by using a cell culture solution containing an inactivated bacterial solution, performing intervention action on the cells for different probiotic strains, washing the cells for 2 times by using a phosphate buffer solution after each group of cells are treated, adding a high-glucose INS-1 complete culture solution with a glucose concentration of 16.7mmol/L, incubating for 0.5-1h at 37 ℃, then collecting cell supernatant, detecting the insulin secretion of each group under high-glucose stimulation by using an ELISA method, performing the ELISA detection step according to a kit instruction, detecting the absorbance at 450nm by using an enzyme labeling instrument, drawing a standard curve by using an OD value as a longitudinal coordinate Y and a standard substance concentration as a horizontal coordinate X, and calculating the concentrations of other samples according to the standard.
12. The method for screening Lactobacillus plantarum according to claim 3, wherein: the construction method of the type 2 diabetes model rat comprises the following steps: after being adaptively fed for 5 days by using basal feed, 70 Wistar rats with 6-8 weeks old male animals are randomly divided into a low-fat group and a high-fat group, and all the groups except the low-fat group are fed with the basal feed for the first four weeks, and the weight measurement record of the rats is carried out at a fixed time every week;
in the fifth week, all rats are fasted for 12-18h without water prohibition, except for a low-fat group, the rats in other groups are injected with newly prepared streptozotocin STZ according to the weight of 30-40mg/kg, a certain amount of STZ is dissolved in a citric acid-sodium citrate buffer solution with the pH value of 4.5, the rats are prepared for use and stored in an ice bath, the low-fat group is injected with a citric acid-sodium citrate buffer solution with the pH value of 4.5, sufficient sterile water and high-fat diet are ensured in 72h after molding, padding is replaced every day, and diabetic rats with random blood sugar higher than 16.7mmol/L or fasting blood sugar higher than 11.7mmol/L after 72h are successfully molded;
dividing the rat into model group, LGG group, and CT152 group, wherein the model group uses 0.85% physiological saline as control, and LGG and CCTCCM2019591 are 1 × 109CFU/day was treated for a 6-8 week period with gavage.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910709658.2A CN110964659A (en) | 2019-08-02 | 2019-08-02 | Lactobacillus plantarum with blood sugar reducing effect, screening method thereof, application medicament and food |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN201910709658.2A CN110964659A (en) | 2019-08-02 | 2019-08-02 | Lactobacillus plantarum with blood sugar reducing effect, screening method thereof, application medicament and food |
Publications (1)
| Publication Number | Publication Date |
|---|---|
| CN110964659A true CN110964659A (en) | 2020-04-07 |
Family
ID=70028382
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN201910709658.2A Pending CN110964659A (en) | 2019-08-02 | 2019-08-02 | Lactobacillus plantarum with blood sugar reducing effect, screening method thereof, application medicament and food |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN110964659A (en) |
Cited By (6)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109022313A (en) * | 2018-08-02 | 2018-12-18 | 中国农业科学院兰州兽医研究所 | One lactobacillus plantarum |
| CN112402465A (en) * | 2020-12-18 | 2021-02-26 | 四川大学华西医院 | Product of lactobacillus plantarum N-1 for preventing and/or improving benign prostatic hyperplasia and application thereof |
| KR20220006864A (en) * | 2020-07-09 | 2022-01-18 | 한국식품연구원 | Lactobacillus plantarum K97 and uses thereof |
| CN114437997A (en) * | 2022-04-07 | 2022-05-06 | 山东向日葵生物工程有限公司 | Lactobacillus plantarum SF-L38 and application thereof in preparation of blood sugar control product |
| CN114752529A (en) * | 2022-04-29 | 2022-07-15 | 科郦有限公司 | Lactobacillus plantarum HOM3201 strain, viable bacteria preparation thereof, preparation method and application |
| CN117535181A (en) * | 2023-10-31 | 2024-02-09 | 四川大学 | Lactobacillus plantarum SC256 and application thereof |
Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105567586A (en) * | 2015-12-21 | 2016-05-11 | 南昌大学 | Lactobacillus plantarum having anti-diabetic function and application thereof |
| CN106834196A (en) * | 2017-04-10 | 2017-06-13 | 吉林省农业科学院 | Lactobacillus plantarum CH126 and its application in function of polysaccharide food is prepared |
| CN107502575A (en) * | 2017-09-20 | 2017-12-22 | 中国农业科学院农产品加工研究所 | One plant of Lactobacillus plantarum with the high inhibitory activity of α glucuroides |
| CN108018236A (en) * | 2017-12-18 | 2018-05-11 | 富乐顿生物工程科技（北京）有限公司 | A kind of spatial plant lactobacillus SS18-37 for reducing alpha-glucosidase activity and its application |
-
2019
- 2019-08-02 CN CN201910709658.2A patent/CN110964659A/en active Pending
Patent Citations (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN105567586A (en) * | 2015-12-21 | 2016-05-11 | 南昌大学 | Lactobacillus plantarum having anti-diabetic function and application thereof |
| CN106834196A (en) * | 2017-04-10 | 2017-06-13 | 吉林省农业科学院 | Lactobacillus plantarum CH126 and its application in function of polysaccharide food is prepared |
| CN107502575A (en) * | 2017-09-20 | 2017-12-22 | 中国农业科学院农产品加工研究所 | One plant of Lactobacillus plantarum with the high inhibitory activity of α glucuroides |
| CN108018236A (en) * | 2017-12-18 | 2018-05-11 | 富乐顿生物工程科技（北京）有限公司 | A kind of spatial plant lactobacillus SS18-37 for reducing alpha-glucosidase activity and its application |
Non-Patent Citations (1)
| Title |
|---|
| TING CAI ET AL.: "In vitro evaluation by PCA and AHP of potential antidiabetic properties of lactic acid bacteria isolated from traditional fermented food", 《LWT - FOOD SCIENCE AND TECHNOLOGY》 * |
Cited By (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN109022313A (en) * | 2018-08-02 | 2018-12-18 | 中国农业科学院兰州兽医研究所 | One lactobacillus plantarum |
| CN109022313B (en) * | 2018-08-02 | 2021-08-24 | 中国农业科学院兰州兽医研究所 | Lactobacillus plantarum |
| KR20220006864A (en) * | 2020-07-09 | 2022-01-18 | 한국식품연구원 | Lactobacillus plantarum K97 and uses thereof |
| KR102570432B1 (en) | 2020-07-09 | 2023-08-25 | 한국식품연구원 | Lactobacillus plantarum K97 and uses thereof |
| CN112402465A (en) * | 2020-12-18 | 2021-02-26 | 四川大学华西医院 | Product of lactobacillus plantarum N-1 for preventing and/or improving benign prostatic hyperplasia and application thereof |
| CN112402465B (en) * | 2020-12-18 | 2022-12-30 | 四川大学华西医院 | Product of lactobacillus plantarum N-1 for preventing and/or improving benign prostatic hyperplasia and application thereof |
| CN114437997A (en) * | 2022-04-07 | 2022-05-06 | 山东向日葵生物工程有限公司 | Lactobacillus plantarum SF-L38 and application thereof in preparation of blood sugar control product |
| CN114437997B (en) * | 2022-04-07 | 2022-06-10 | 山东向日葵生物工程有限公司 | Lactobacillus plantarum SF-L38 and application thereof in preparation of blood sugar control product |
| CN114752529A (en) * | 2022-04-29 | 2022-07-15 | 科郦有限公司 | Lactobacillus plantarum HOM3201 strain, viable bacteria preparation thereof, preparation method and application |
| CN114752529B (en) * | 2022-04-29 | 2023-12-19 | 科郦有限公司 | Lactobacillus plantarum HOM3201 strain and viable bacteria preparation, preparation method and application thereof |
| CN117535181A (en) * | 2023-10-31 | 2024-02-09 | 四川大学 | Lactobacillus plantarum SC256 and application thereof |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN110964659A (en) | Lactobacillus plantarum with blood sugar reducing effect, screening method thereof, application medicament and food | |
| CN110157647B (en) | Lactobacillus brevis capable of relieving anxiety and improving sleep and application thereof | |
| CN110835616B (en) | Active substance of lactobacillus paracasei GKS6, composition containing same and application of active substance in promoting longevity | |
| EP3715448A2 (en) | Novel lactic acid bacteria and use thereof | |
| CN109182184B (en) | Pediococcus acidilactici strain and application thereof | |
| CN109182162B (en) | Lactobacillus plantarum with antioxidant capacity and application thereof | |
| CA2609617A1 (en) | Feline probiotic lactobacilli | |
| CN109234189A (en) | One plant of lactobacillus plantarum strain BX62 and its application with oxidation resistance | |
| CN113308421B (en) | Lactobacillus plantarum BUFX and application thereof in metabolic syndrome | |
| CN116064326B (en) | Bifidobacterium animalis subspecies GBW8051 capable of relieving depression and application thereof | |
| CN114574406B (en) | Lactobacillus rhamnosus strain WKA55, and application and product thereof in preparation of product for preventing and treating alcoholic liver injury | |
| CN110577912A (en) | lactobacillus gasseri and application thereof in preparing fermented milk | |
| CN110833565B (en) | Active substance of lactobacillus plantarum GKM3, composition containing same and application of active substance in promoting longevity | |
| CN112708574B (en) | Lactobacillus bulgaricus TCI904, compositions thereof and use thereof for reducing body weight | |
| Garcia-Mazcorro et al. | Are there indigenous Saccharomyces in the digestive tract of livestock animal species? Implications for health, nutrition and productivity traits | |
| CN115322932B (en) | Lactobacillus plantarum with anti-alcohol and sobering-up capabilities and application thereof | |
| CN112154202A (en) | Simulating an intestinal environment | |
| CN113322216A (en) | Lactobacillus paracasei B111H and application thereof in metabolic syndrome | |
| JP2024532769A (en) | Postbiotics | |
| CN113444656B (en) | Strain with anti-aging function and application thereof | |
| Aklilu et al. | Identification and biotyping of Escherichia coli from diarrheic lambs in and around Debre Birhan town, Ethiopia | |
| TWI669124B (en) | Use of bacillus coagulans for preparing a pharmaceutical composition for reducing levels of heavy metals and protecting the liver | |
| Bobga et al. | Evaluation of the anti-diabetic potential of probiotic Lactobacillus fermentum (PRI 29) isolated from cameroonian fermented cow milk in alloxan induced diabetes type-1 mice model | |
| CN115960784A (en) | Lactobacillus plantarum ZJUF SYS1 and application thereof | |
| CA3085995A1 (en) | Composition for type i allergy |
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 | ||
| RJ01 | Rejection of invention patent application after publication | ||
| RJ01 | Rejection of invention patent application after publication |
Application publication date: 20200407 |