CN116790448A - Lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof - Google Patents
Lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof Download PDFInfo
- Publication number
- CN116790448A CN116790448A CN202311051207.7A CN202311051207A CN116790448A CN 116790448 A CN116790448 A CN 116790448A CN 202311051207 A CN202311051207 A CN 202311051207A CN 116790448 A CN116790448 A CN 116790448A
- Authority
- CN
- China
- Prior art keywords
- opb102
- lactobacillus helveticus
- diarrhea
- constipation
- group
- 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.)
- Granted
Links
- 240000002605 Lactobacillus helveticus Species 0.000 title claims abstract description 96
- 235000013967 Lactobacillus helveticus Nutrition 0.000 title claims abstract description 96
- 229940054346 lactobacillus helveticus Drugs 0.000 title claims abstract description 96
- 206010012735 Diarrhoea Diseases 0.000 title claims abstract description 88
- 206010010774 Constipation Diseases 0.000 title claims abstract description 69
- 210000001072 colon Anatomy 0.000 claims abstract description 23
- 239000003814 drug Substances 0.000 claims abstract description 17
- 210000001519 tissue Anatomy 0.000 claims abstract description 16
- 235000013305 food Nutrition 0.000 claims abstract description 13
- 230000002496 gastric effect Effects 0.000 claims abstract description 11
- 210000002175 goblet cell Anatomy 0.000 claims abstract description 11
- 238000009629 microbiological culture Methods 0.000 claims abstract description 6
- 238000004321 preservation Methods 0.000 claims abstract description 6
- 239000003795 chemical substances by application Substances 0.000 claims abstract description 5
- 230000000968 intestinal effect Effects 0.000 claims description 37
- 241000894006 Bacteria Species 0.000 claims description 27
- 230000000694 effects Effects 0.000 claims description 22
- 230000001965 increasing effect Effects 0.000 claims description 16
- 210000000813 small intestine Anatomy 0.000 claims description 14
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 13
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 9
- 229920002472 Starch Polymers 0.000 claims description 9
- 238000002360 preparation method Methods 0.000 claims description 9
- 239000008107 starch Substances 0.000 claims description 9
- 235000019698 starch Nutrition 0.000 claims description 9
- 230000009286 beneficial effect Effects 0.000 claims description 7
- 230000013872 defecation Effects 0.000 claims description 7
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 6
- 210000004027 cell Anatomy 0.000 claims description 6
- 239000003085 diluting agent Substances 0.000 claims description 6
- 239000000945 filler Substances 0.000 claims description 6
- 239000012530 fluid Substances 0.000 claims description 6
- 239000000314 lubricant Substances 0.000 claims description 6
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 claims description 6
- 244000005700 microbiome Species 0.000 claims description 6
- 241001465754 Metazoa Species 0.000 claims description 5
- 230000012010 growth Effects 0.000 claims description 5
- 239000001963 growth medium Substances 0.000 claims description 5
- 238000012258 culturing Methods 0.000 claims description 4
- 239000003937 drug carrier Substances 0.000 claims description 4
- 230000002550 fecal effect Effects 0.000 claims description 4
- 230000002401 inhibitory effect Effects 0.000 claims description 4
- 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 claims description 3
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims description 3
- 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 claims description 3
- 239000004375 Dextrin Substances 0.000 claims description 3
- 229920001353 Dextrin Polymers 0.000 claims description 3
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Chemical compound OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims description 3
- GUBGYTABKSRVRQ-QKKXKWKRSA-N Lactose Natural products OC[C@H]1O[C@@H](O[C@H]2[C@H](O)[C@@H](O)C(O)O[C@@H]2CO)[C@H](O)[C@@H](O)[C@H]1O GUBGYTABKSRVRQ-QKKXKWKRSA-N 0.000 claims description 3
- 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 claims description 3
- 229930195725 Mannitol Natural products 0.000 claims description 3
- 229920000168 Microcrystalline cellulose Polymers 0.000 claims description 3
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 claims description 3
- 239000000853 adhesive Substances 0.000 claims description 3
- 230000001070 adhesive effect Effects 0.000 claims description 3
- 230000001668 ameliorated effect Effects 0.000 claims description 3
- 239000011230 binding agent Substances 0.000 claims description 3
- 230000004071 biological effect Effects 0.000 claims description 3
- 235000019425 dextrin Nutrition 0.000 claims description 3
- FPAFDBFIGPHWGO-UHFFFAOYSA-N dioxosilane;oxomagnesium;hydrate Chemical compound O.[Mg]=O.[Mg]=O.[Mg]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O FPAFDBFIGPHWGO-UHFFFAOYSA-N 0.000 claims description 3
- 239000007884 disintegrant Substances 0.000 claims description 3
- 235000021107 fermented food Nutrition 0.000 claims description 3
- 235000013402 health food Nutrition 0.000 claims description 3
- 235000012907 honey Nutrition 0.000 claims description 3
- 239000008101 lactose Substances 0.000 claims description 3
- 229940031703 low substituted hydroxypropyl cellulose Drugs 0.000 claims description 3
- 235000019359 magnesium stearate Nutrition 0.000 claims description 3
- 239000000594 mannitol Substances 0.000 claims description 3
- 235000010355 mannitol Nutrition 0.000 claims description 3
- 239000000463 material Substances 0.000 claims description 3
- 239000002207 metabolite Substances 0.000 claims description 3
- 239000008108 microcrystalline cellulose Substances 0.000 claims description 3
- 235000019813 microcrystalline cellulose Nutrition 0.000 claims description 3
- 229940016286 microcrystalline cellulose Drugs 0.000 claims description 3
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims description 3
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims description 3
- 229940069328 povidone Drugs 0.000 claims description 3
- 230000001737 promoting effect Effects 0.000 claims description 3
- 238000004904 shortening Methods 0.000 claims description 3
- 239000000377 silicon dioxide Substances 0.000 claims description 3
- 235000012239 silicon dioxide Nutrition 0.000 claims description 3
- 239000011734 sodium Substances 0.000 claims description 3
- 229910052708 sodium Inorganic materials 0.000 claims description 3
- OKODKVMXHLUQSW-JITBQSAISA-M sodium;(e)-4-hydroxy-4-oxobut-2-enoate;octadecanoic acid Chemical compound [Na+].OC(=O)\C=C\C([O-])=O.CCCCCCCCCCCCCCCCCC(O)=O OKODKVMXHLUQSW-JITBQSAISA-M 0.000 claims description 3
- 239000006188 syrup Substances 0.000 claims description 3
- 235000020357 syrup Nutrition 0.000 claims description 3
- 238000000855 fermentation Methods 0.000 claims 1
- 230000004151 fermentation Effects 0.000 claims 1
- 229940079593 drug Drugs 0.000 abstract description 3
- 230000000813 microbial effect Effects 0.000 abstract description 2
- 241000699670 Mus sp. Species 0.000 description 53
- 239000006041 probiotic Substances 0.000 description 34
- 235000018291 probiotics Nutrition 0.000 description 34
- 230000000529 probiotic effect Effects 0.000 description 19
- 239000000203 mixture Substances 0.000 description 17
- 241000186660 Lactobacillus Species 0.000 description 14
- 229940039696 lactobacillus Drugs 0.000 description 14
- 239000000243 solution Substances 0.000 description 12
- 238000011282 treatment Methods 0.000 description 11
- 239000013641 positive control Substances 0.000 description 9
- 239000000047 product Substances 0.000 description 9
- 241000894007 species Species 0.000 description 9
- 238000012360 testing method Methods 0.000 description 9
- 241000869429 Muribaculaceae Species 0.000 description 8
- 238000001179 sorption measurement Methods 0.000 description 8
- 239000000725 suspension Substances 0.000 description 8
- 230000001580 bacterial effect Effects 0.000 description 7
- 244000052616 bacterial pathogen Species 0.000 description 7
- 210000001035 gastrointestinal tract Anatomy 0.000 description 7
- 210000004347 intestinal mucosa Anatomy 0.000 description 7
- 230000002829 reductive effect Effects 0.000 description 7
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 7
- 230000008859 change Effects 0.000 description 6
- 230000029087 digestion Effects 0.000 description 6
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 6
- 239000007788 liquid Substances 0.000 description 6
- 230000001105 regulatory effect Effects 0.000 description 6
- 240000001046 Lactobacillus acidophilus Species 0.000 description 5
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 5
- 230000033228 biological regulation Effects 0.000 description 5
- 239000012153 distilled water Substances 0.000 description 5
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 5
- 230000007774 longterm Effects 0.000 description 5
- 238000000034 method Methods 0.000 description 5
- 238000002156 mixing Methods 0.000 description 5
- 238000010186 staining Methods 0.000 description 5
- 241000186000 Bifidobacterium Species 0.000 description 4
- 208000036649 Dysbacteriosis Diseases 0.000 description 4
- 208000027244 Dysbiosis Diseases 0.000 description 4
- 241000699666 Mus <mouse, genus> Species 0.000 description 4
- 241000566145 Otus Species 0.000 description 4
- 238000004458 analytical method Methods 0.000 description 4
- 230000002457 bidirectional effect Effects 0.000 description 4
- 230000003115 biocidal effect Effects 0.000 description 4
- 230000007140 dysbiosis Effects 0.000 description 4
- 238000002474 experimental method Methods 0.000 description 4
- 235000011389 fruit/vegetable juice Nutrition 0.000 description 4
- 230000006870 function Effects 0.000 description 4
- 229960002983 loperamide hydrochloride Drugs 0.000 description 4
- PGYPOBZJRVSMDS-UHFFFAOYSA-N loperamide hydrochloride Chemical compound Cl.C=1C=CC=CC=1C(C=1C=CC=CC=1)(C(=O)N(C)C)CCN(CC1)CCC1(O)C1=CC=C(Cl)C=C1 PGYPOBZJRVSMDS-UHFFFAOYSA-N 0.000 description 4
- 235000010482 polyoxyethylene sorbitan monooleate Nutrition 0.000 description 4
- 229920000053 polysorbate 80 Polymers 0.000 description 4
- 230000004083 survival effect Effects 0.000 description 4
- 239000003981 vehicle Substances 0.000 description 4
- 208000004232 Enteritis Diseases 0.000 description 3
- 241001112693 Lachnospiraceae Species 0.000 description 3
- 241001468155 Lactobacillaceae Species 0.000 description 3
- 240000006024 Lactobacillus plantarum Species 0.000 description 3
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 3
- 239000007864 aqueous solution Substances 0.000 description 3
- 238000006243 chemical reaction Methods 0.000 description 3
- 230000034994 death Effects 0.000 description 3
- 235000019441 ethanol Nutrition 0.000 description 3
- 210000004051 gastric juice Anatomy 0.000 description 3
- 238000000338 in vitro Methods 0.000 description 3
- 235000014655 lactic acid Nutrition 0.000 description 3
- 239000004310 lactic acid Substances 0.000 description 3
- 229940072205 lactobacillus plantarum Drugs 0.000 description 3
- 239000011259 mixed solution Substances 0.000 description 3
- 230000008855 peristalsis Effects 0.000 description 3
- 238000012163 sequencing technique Methods 0.000 description 3
- 235000020183 skimmed milk Nutrition 0.000 description 3
- 210000002784 stomach Anatomy 0.000 description 3
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- 238000007400 DNA extraction Methods 0.000 description 2
- WSFSSNUMVMOOMR-UHFFFAOYSA-N Formaldehyde Chemical compound O=C WSFSSNUMVMOOMR-UHFFFAOYSA-N 0.000 description 2
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Natural products OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 2
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 2
- 241001052560 Thallis Species 0.000 description 2
- 201000009840 acute diarrhea Diseases 0.000 description 2
- 230000001154 acute effect Effects 0.000 description 2
- 239000003242 anti bacterial agent Substances 0.000 description 2
- 229940088710 antibiotic agent Drugs 0.000 description 2
- 230000008901 benefit Effects 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 238000004364 calculation method Methods 0.000 description 2
- 239000004359 castor oil Substances 0.000 description 2
- 235000019438 castor oil Nutrition 0.000 description 2
- 230000000052 comparative effect Effects 0.000 description 2
- 230000007547 defect Effects 0.000 description 2
- 238000004925 denaturation Methods 0.000 description 2
- 230000036425 denaturation Effects 0.000 description 2
- 238000010586 diagram Methods 0.000 description 2
- 238000010790 dilution Methods 0.000 description 2
- 239000012895 dilution Substances 0.000 description 2
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- 235000013376 functional food Nutrition 0.000 description 2
- ZEMPKEQAKRGZGQ-XOQCFJPHSA-N glycerol triricinoleate Natural products CCCCCC[C@@H](O)CC=CCCCCCCCC(=O)OC[C@@H](COC(=O)CCCCCCCC=CC[C@@H](O)CCCCCC)OC(=O)CCCCCCCC=CC[C@H](O)CCCCCC ZEMPKEQAKRGZGQ-XOQCFJPHSA-N 0.000 description 2
- 230000002008 hemorrhagic effect Effects 0.000 description 2
- 230000001976 improved effect Effects 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 210000003097 mucus Anatomy 0.000 description 2
- 210000003205 muscle Anatomy 0.000 description 2
- 230000001575 pathological effect Effects 0.000 description 2
- 239000008057 potassium phosphate buffer Substances 0.000 description 2
- 239000002244 precipitate Substances 0.000 description 2
- 238000001556 precipitation Methods 0.000 description 2
- 230000002265 prevention Effects 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 229940124513 senna glycoside Drugs 0.000 description 2
- 239000000126 substance Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 210000004876 tela submucosa Anatomy 0.000 description 2
- 231100000331 toxic Toxicity 0.000 description 2
- 230000002588 toxic effect Effects 0.000 description 2
- 230000000007 visual effect Effects 0.000 description 2
- 239000002699 waste material Substances 0.000 description 2
- 235000013618 yogurt Nutrition 0.000 description 2
- 108020004465 16S ribosomal RNA Proteins 0.000 description 1
- QKNYBSVHEMOAJP-UHFFFAOYSA-N 2-amino-2-(hydroxymethyl)propane-1,3-diol;hydron;chloride Chemical compound Cl.OCC(N)(CO)CO QKNYBSVHEMOAJP-UHFFFAOYSA-N 0.000 description 1
- 208000004998 Abdominal Pain Diseases 0.000 description 1
- 206010000060 Abdominal distension Diseases 0.000 description 1
- 229920001817 Agar Polymers 0.000 description 1
- 241000186018 Bifidobacterium adolescentis Species 0.000 description 1
- 241001608472 Bifidobacterium longum Species 0.000 description 1
- 206010007134 Candida infections Diseases 0.000 description 1
- CURLTUGMZLYLDI-UHFFFAOYSA-N Carbon dioxide Chemical compound O=C=O CURLTUGMZLYLDI-UHFFFAOYSA-N 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 244000025596 Cassia laevigata Species 0.000 description 1
- 235000006693 Cassia laevigata Nutrition 0.000 description 1
- 102000016938 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- VWDXGKUTGQJJHJ-UHFFFAOYSA-N Catenarin Natural products C1=C(O)C=C2C(=O)C3=C(O)C(C)=CC(O)=C3C(=O)C2=C1O VWDXGKUTGQJJHJ-UHFFFAOYSA-N 0.000 description 1
- 241000207199 Citrus Species 0.000 description 1
- KCXVZYZYPLLWCC-UHFFFAOYSA-N EDTA Chemical compound OC(=O)CN(CC(O)=O)CCN(CC(O)=O)CC(O)=O KCXVZYZYPLLWCC-UHFFFAOYSA-N 0.000 description 1
- 239000010282 Emodin Substances 0.000 description 1
- RBLJKYCRSCQLRP-UHFFFAOYSA-N Emodin-dianthron Natural products O=C1C2=CC(C)=CC(O)=C2C(=O)C2=C1CC(=O)C=C2O RBLJKYCRSCQLRP-UHFFFAOYSA-N 0.000 description 1
- 108010067770 Endopeptidase K Proteins 0.000 description 1
- 241000194032 Enterococcus faecalis Species 0.000 description 1
- 108090000790 Enzymes Proteins 0.000 description 1
- 102000004190 Enzymes Human genes 0.000 description 1
- 241000588724 Escherichia coli Species 0.000 description 1
- 206010061126 Escherichia infection Diseases 0.000 description 1
- 241001289529 Fallopia multiflora Species 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 238000003794 Gram staining Methods 0.000 description 1
- YOOXNSPYGCZLAX-UHFFFAOYSA-N Helminthosporin Natural products C1=CC(O)=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O YOOXNSPYGCZLAX-UHFFFAOYSA-N 0.000 description 1
- 229920001202 Inulin Polymers 0.000 description 1
- 241000186605 Lactobacillus paracasei Species 0.000 description 1
- 206010067125 Liver injury Diseases 0.000 description 1
- 102000016943 Muramidase Human genes 0.000 description 1
- 108010014251 Muramidase Proteins 0.000 description 1
- 108010062010 N-Acetylmuramoyl-L-alanine Amidase Proteins 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- 241000692844 Prevotellaceae Species 0.000 description 1
- 208000003100 Pseudomembranous Enterocolitis Diseases 0.000 description 1
- NTGIIKCGBNGQAR-UHFFFAOYSA-N Rheoemodin Natural products C1=C(O)C=C2C(=O)C3=CC(O)=CC(O)=C3C(=O)C2=C1O NTGIIKCGBNGQAR-UHFFFAOYSA-N 0.000 description 1
- 244000299790 Rheum rhabarbarum Species 0.000 description 1
- 235000009411 Rheum rhabarbarum Nutrition 0.000 description 1
- VMHLLURERBWHNL-UHFFFAOYSA-M Sodium acetate Chemical compound [Na+].CC([O-])=O VMHLLURERBWHNL-UHFFFAOYSA-M 0.000 description 1
- UQZIYBXSHAGNOE-USOSMYMVSA-N Stachyose Natural products O(C[C@H]1[C@@H](O)[C@H](O)[C@H](O)[C@@H](O[C@@]2(CO)[C@H](O)[C@@H](O)[C@@H](CO)O2)O1)[C@@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@H](CO[C@@H]2[C@@H](O)[C@@H](O)[C@@H](O)[C@H](CO)O2)O1 UQZIYBXSHAGNOE-USOSMYMVSA-N 0.000 description 1
- 102000000591 Tight Junction Proteins Human genes 0.000 description 1
- 108010002321 Tight Junction Proteins Proteins 0.000 description 1
- 239000008272 agar Substances 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 238000000246 agarose gel electrophoresis Methods 0.000 description 1
- 238000000137 annealing Methods 0.000 description 1
- PYKYMHQGRFAEBM-UHFFFAOYSA-N anthraquinone Natural products CCC(=O)c1c(O)c2C(=O)C3C(C=CC=C3O)C(=O)c2cc1CC(=O)OC PYKYMHQGRFAEBM-UHFFFAOYSA-N 0.000 description 1
- 150000004056 anthraquinones Chemical class 0.000 description 1
- 230000001142 anti-diarrhea Effects 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 229940009291 bifidobacterium longum Drugs 0.000 description 1
- 239000003833 bile salt Substances 0.000 description 1
- 238000007664 blowing Methods 0.000 description 1
- 230000037396 body weight Effects 0.000 description 1
- 201000003984 candidiasis Diseases 0.000 description 1
- 235000011089 carbon dioxide Nutrition 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 238000002512 chemotherapy Methods 0.000 description 1
- 208000019902 chronic diarrheal disease Diseases 0.000 description 1
- 235000020971 citrus fruits Nutrition 0.000 description 1
- 238000010224 classification analysis Methods 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 238000007405 data analysis Methods 0.000 description 1
- 230000003247 decreasing effect Effects 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 201000010099 disease Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 230000004064 dysfunction Effects 0.000 description 1
- 235000006694 eating habits Nutrition 0.000 description 1
- 239000003480 eluent Substances 0.000 description 1
- RHMXXJGYXNZAPX-UHFFFAOYSA-N emodin Chemical compound C1=C(O)C=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O RHMXXJGYXNZAPX-UHFFFAOYSA-N 0.000 description 1
- VASFLQKDXBAWEL-UHFFFAOYSA-N emodin Natural products OC1=C(OC2=C(C=CC(=C2C1=O)O)O)C1=CC=C(C=C1)O VASFLQKDXBAWEL-UHFFFAOYSA-N 0.000 description 1
- 238000005516 engineering process Methods 0.000 description 1
- 230000002708 enhancing effect Effects 0.000 description 1
- 229940032049 enterococcus faecalis Drugs 0.000 description 1
- 238000003912 environmental pollution Methods 0.000 description 1
- 229940088598 enzyme Drugs 0.000 description 1
- 208000020612 escherichia coli infection Diseases 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 239000000835 fiber Substances 0.000 description 1
- 229940050549 fiber Drugs 0.000 description 1
- 235000013373 food additive Nutrition 0.000 description 1
- 239000002778 food additive Substances 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
- 239000000499 gel Substances 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 230000036541 health Effects 0.000 description 1
- 238000007490 hematoxylin and eosin (H&E) staining Methods 0.000 description 1
- 231100000753 hepatic injury Toxicity 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 208000028774 intestinal disease Diseases 0.000 description 1
- 230000003870 intestinal permeability Effects 0.000 description 1
- JYJIGFIDKWBXDU-MNNPPOADSA-N inulin Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)OC[C@]1(OC[C@]2(OC[C@]3(OC[C@]4(OC[C@]5(OC[C@]6(OC[C@]7(OC[C@]8(OC[C@]9(OC[C@]%10(OC[C@]%11(OC[C@]%12(OC[C@]%13(OC[C@]%14(OC[C@]%15(OC[C@]%16(OC[C@]%17(OC[C@]%18(OC[C@]%19(OC[C@]%20(OC[C@]%21(OC[C@]%22(OC[C@]%23(OC[C@]%24(OC[C@]%25(OC[C@]%26(OC[C@]%27(OC[C@]%28(OC[C@]%29(OC[C@]%30(OC[C@]%31(OC[C@]%32(OC[C@]%33(OC[C@]%34(OC[C@]%35(OC[C@]%36(O[C@@H]%37[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O%37)O)[C@H]([C@H](O)[C@@H](CO)O%36)O)[C@H]([C@H](O)[C@@H](CO)O%35)O)[C@H]([C@H](O)[C@@H](CO)O%34)O)[C@H]([C@H](O)[C@@H](CO)O%33)O)[C@H]([C@H](O)[C@@H](CO)O%32)O)[C@H]([C@H](O)[C@@H](CO)O%31)O)[C@H]([C@H](O)[C@@H](CO)O%30)O)[C@H]([C@H](O)[C@@H](CO)O%29)O)[C@H]([C@H](O)[C@@H](CO)O%28)O)[C@H]([C@H](O)[C@@H](CO)O%27)O)[C@H]([C@H](O)[C@@H](CO)O%26)O)[C@H]([C@H](O)[C@@H](CO)O%25)O)[C@H]([C@H](O)[C@@H](CO)O%24)O)[C@H]([C@H](O)[C@@H](CO)O%23)O)[C@H]([C@H](O)[C@@H](CO)O%22)O)[C@H]([C@H](O)[C@@H](CO)O%21)O)[C@H]([C@H](O)[C@@H](CO)O%20)O)[C@H]([C@H](O)[C@@H](CO)O%19)O)[C@H]([C@H](O)[C@@H](CO)O%18)O)[C@H]([C@H](O)[C@@H](CO)O%17)O)[C@H]([C@H](O)[C@@H](CO)O%16)O)[C@H]([C@H](O)[C@@H](CO)O%15)O)[C@H]([C@H](O)[C@@H](CO)O%14)O)[C@H]([C@H](O)[C@@H](CO)O%13)O)[C@H]([C@H](O)[C@@H](CO)O%12)O)[C@H]([C@H](O)[C@@H](CO)O%11)O)[C@H]([C@H](O)[C@@H](CO)O%10)O)[C@H]([C@H](O)[C@@H](CO)O9)O)[C@H]([C@H](O)[C@@H](CO)O8)O)[C@H]([C@H](O)[C@@H](CO)O7)O)[C@H]([C@H](O)[C@@H](CO)O6)O)[C@H]([C@H](O)[C@@H](CO)O5)O)[C@H]([C@H](O)[C@@H](CO)O4)O)[C@H]([C@H](O)[C@@H](CO)O3)O)[C@H]([C@H](O)[C@@H](CO)O2)O)[C@@H](O)[C@H](O)[C@@H](CO)O1 JYJIGFIDKWBXDU-MNNPPOADSA-N 0.000 description 1
- 229940029339 inulin Drugs 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- 208000002551 irritable bowel syndrome Diseases 0.000 description 1
- 238000002955 isolation Methods 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 239000000832 lactitol Substances 0.000 description 1
- 235000010448 lactitol Nutrition 0.000 description 1
- VQHSOMBJVWLPSR-JVCRWLNRSA-N lactitol Chemical compound OC[C@H](O)[C@@H](O)[C@@H]([C@H](O)CO)O[C@@H]1O[C@H](CO)[C@H](O)[C@H](O)[C@H]1O VQHSOMBJVWLPSR-JVCRWLNRSA-N 0.000 description 1
- 229960003451 lactitol Drugs 0.000 description 1
- 229940068140 lactobacillus bifidus Drugs 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 230000033001 locomotion Effects 0.000 description 1
- 239000004325 lysozyme Substances 0.000 description 1
- 229960000274 lysozyme Drugs 0.000 description 1
- 235000010335 lysozyme Nutrition 0.000 description 1
- 239000002075 main ingredient Substances 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 238000005259 measurement Methods 0.000 description 1
- 230000006996 mental state Effects 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- 210000004877 mucosa Anatomy 0.000 description 1
- 230000004682 mucosal barrier function Effects 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 230000003287 optical effect Effects 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 239000002245 particle Substances 0.000 description 1
- 231100000915 pathological change Toxicity 0.000 description 1
- 230000036285 pathological change Effects 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- PKUBGLYEOAJPEG-UHFFFAOYSA-N physcion Natural products C1=C(C)C=C2C(=O)C3=CC(C)=CC(O)=C3C(=O)C2=C1O PKUBGLYEOAJPEG-UHFFFAOYSA-N 0.000 description 1
- 231100000614 poison Toxicity 0.000 description 1
- 230000008092 positive effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 238000000746 purification Methods 0.000 description 1
- 210000001187 pylorus Anatomy 0.000 description 1
- 239000002994 raw material Substances 0.000 description 1
- 238000011160 research Methods 0.000 description 1
- 230000004044 response Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 230000000630 rising effect Effects 0.000 description 1
- 229960002181 saccharomyces boulardii Drugs 0.000 description 1
- 229930186851 sennoside Natural products 0.000 description 1
- IPQVTOJGNYVQEO-KGFNBKMBSA-N sennoside A Chemical compound O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@H]1OC1=CC=CC2=C1C(=O)C1=C(O)C=C(C(O)=O)C=C1[C@@H]2[C@H]1C2=CC(C(O)=O)=CC(O)=C2C(=O)C2=C(O[C@H]3[C@@H]([C@@H](O)[C@H](O)[C@@H](CO)O3)O)C=CC=C21 IPQVTOJGNYVQEO-KGFNBKMBSA-N 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 239000011780 sodium chloride Substances 0.000 description 1
- UQZIYBXSHAGNOE-XNSRJBNMSA-N stachyose Chemical compound O[C@H]1[C@H](O)[C@@H](CO)O[C@@]1(CO)O[C@@H]1[C@H](O)[C@@H](O)[C@H](O)[C@@H](CO[C@@H]2[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO[C@@H]3[C@@H]([C@@H](O)[C@@H](O)[C@@H](CO)O3)O)O2)O)O1 UQZIYBXSHAGNOE-XNSRJBNMSA-N 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 239000006228 supernatant Substances 0.000 description 1
- 230000001502 supplementing effect Effects 0.000 description 1
- 210000001578 tight junction Anatomy 0.000 description 1
- 239000003440 toxic substance Substances 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 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
- 238000005406 washing Methods 0.000 description 1
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
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
- A61P1/10—Laxatives
-
- 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
- A61P1/12—Antidiarrhoeals
-
- 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
-
- 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
-
- 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
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Organic Chemistry (AREA)
- Medicinal Chemistry (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Veterinary Medicine (AREA)
- Public Health (AREA)
- Animal Behavior & Ethology (AREA)
- Microbiology (AREA)
- Pharmacology & Pharmacy (AREA)
- Mycology (AREA)
- General Chemical & Material Sciences (AREA)
- Genetics & Genomics (AREA)
- Biotechnology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Zoology (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Wood Science & Technology (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Biomedical Technology (AREA)
- Virology (AREA)
- General Engineering & Computer Science (AREA)
- Tropical Medicine & Parasitology (AREA)
- Biochemistry (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
The invention belongs to the technical field of biology, and discloses lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof, wherein the name is: OPB102, class name: lactobacillus helveticus with deposit No.: CGMCC No.26806, preservation unit: china general microbiological culture Collection center (China Committee for culture Collection). The OPB102 can endure the gastrointestinal environment, and can be used for preparing functional microbial agents, foods and medicines for relieving constipation or diarrhea; at the same time, it can restore damaged colon tissue, improve villus density and increase goblet cell number.
Description
Technical Field
The invention belongs to the technical field of biology, and particularly relates to lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof.
Background
Constipation is a common intestinal disease, mainly manifested by difficult defecation, dry and hard feces and less than 3 times per week, and the incidence rate of constipation tends to rise year by year. Short-term constipation affects normal work and life of people, long-term constipation can even cause kidney diseases and cardiovascular diseases, and therefore people should pay attention to prevention and treatment of constipation. Constipation is affected by a variety of factors including changes in life and eating habits, changes in mental state, etc. Research shows that intestinal flora in intestinal tracts of constipation patients is disturbed, bifidobacteria and lactobacillus are reduced, and pathogenic bacteria and conditional pathogenic bacteria are increased. This suggests that we can regulate intestinal flora by supplementing probiotics in vitro, so as to restore normal intestinal movement and achieve the purpose of relieving constipation.
Diarrhea is also a common bowel disorder, as opposed to constipation. As environmental pollution is increasing, the use of chemical food additives, frequent international trips, and antibiotic treatment and chemotherapy, results in 5% of patients suffering from diarrhea each year. Of these, 25% of patients are faced with antibiotic-induced diarrhea problems due to long-term antibiotic treatment, and treatment of antibiotic-associated diarrhea with probiotics has demonstrated significant effects of probiotics on the regulation of intestinal microbiota balance. Another study showed that the use of mixed cultures of Lactobacillus acidophilus, bifidobacterium and Saccharomyces boulardii had a good effect on traveler diarrhea prevention. Likewise, the use of probiotic lactic acid bacteria and bifidobacteria may reduce the time of acute diarrhea in children by 30% to 40%.
Probiotics and intestinal tract regulating function. Probiotics are essential elements for human health, and can synthesize various vitamins, participate in digestion of food, promote intestinal peristalsis, inhibit growth of pathogenic bacteria, decompose harmful and toxic substances, etc. The probiotics can directly or indirectly regulate intestinal flora composition of human body, improve mucosa barrier integrity, and increase intestinal probiotic amount in symbiotic or symbiotic relationship, and can be used for treating diarrhea, constipation or irritable bowel syndrome.
Diarrhea and constipation caused by unbalanced intestinal flora, and the existing treatment methods mainly comprise antibiotic adjustment, traditional Chinese medicine adjustment and probiotic preparation adjustment, but have certain defects. First, antibiotics regulate. The antibiotics are taken for a long time, so that dysbacteriosis is more easily caused, and even double infection such as pseudomembranous enteritis, acute hemorrhagic enteritis, candida infection and the like, and acute hemorrhagic enteritis and the like are caused. Second, traditional Chinese medicine is regulated. The traditional Chinese medicine for treating constipation and diarrhea has a plurality of side effects after long-term administration, for example, the traditional Chinese medicine for treating constipation usually contains traditional Chinese medicines such as rhubarb, senna leaf, polygonum multiflorum and the like, and contains small molecular components such as sennoside, emodin, anthraquinone and the like, and the traditional Chinese medicine has quick response, but can cause further dysbacteriosis, aggravate constipation, even produce abdominal pain, black bowel disease, liver injury and other side effects after long-term administration. Third, probiotic preparation modulation. At present, the probiotic preparation is used for regulating diarrhea and constipation, most of the products are multi-strain mixed products, and the effect of regulating diarrhea and constipation at the same time is not obvious. The main components of the multi-strain mixed product, such as the Xinyi pefeikang, are bifidobacterium longum, lactobacillus acidophilus and enterococcus faecalis, are used for treating diarrhea and abdominal distension caused by intestinal dysbacteriosis, and are mainly used for treating diarrhea and poor constipation. A single strain product, such as Lizhu Changle, contains Bifidobacterium adolescentis as main ingredient, and is used for treating intestinal dysfunction caused by intestinal dysbacteriosis, such as acute diarrhea, chronic diarrhea, constipation, etc., but has insignificant overall bidirectional regulation effect.
Chinese patent publication No.: CN112210516a, publication date: 2021, 1 and 12 days, a lactobacillus helveticus L1258 with intestinal tract regulating function and a composition thereof are disclosed. The composition mainly comprises an A component (lactobacillus helveticus L1258, lactobacillus acidophilus La28 and lactobacillus plantarum LP 45) and a B component (citrus fiber, inulin, lactitol and stachyose). 150 people are selected for clinical trial taking, and the age is 18-60 years; the composition meets the Roman III standard of constipation, the total effective rate of the composition is 66.67-80% after being taken for 7 days, and the total effective rate of the composition is 73.33-96.67% after being taken for 14 days. In addition, diarrhea was treated with the E.coli EAEC infection diarrhea model, and 100 mice (5 weeks old) were randomly grouped into 10 groups, with reduced average time to cure the first diarrhea and reduced incidence of re-diarrhea. The above patent publications only studied the human constipation and diarrhea model of escherichia coli infection in mice on lactobacillus helveticus L1258, lactobacillus acidophilus La28 and lactobacillus plantarum LP45 compositions, and did not study lactobacillus helveticus L1258 alone and comprehensively for constipation and diarrhea.
The invention mainly adopts a single strain, namely lactobacillus helveticus OPB102, and simultaneously carries out bidirectional regulation on diarrhea and constipation, and through searching, no related patent publication document for carrying out bidirectional regulation on diarrhea and constipation by using the single strain lactobacillus helveticus is found.
Disclosure of Invention
The invention aims to overcome the defects of the prior art and provides lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof.
The technical scheme adopted for solving the technical problems is as follows:
lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea is named: OPB102, class name: lactobacillus helveticus (Lactobacillus helveticus), accession number: CGMCC No.26806, date of preservation: 2023, 2, 14, deposit unit: china general microbiological culture Collection center (CGMCC) of the China Committee for culture Collection of microorganisms, beijing, chaoyang area, north Chenxi Lu No. 1, 3.
Further, the lactobacillus helveticus OPB102 has the following biological properties:
(1) Characteristics of the cells: gram-positive, non-sporulating, non-motile bacteria;
(2) Colony characteristics: the convex part is smooth and neat;
(3) Growth characteristics: culturing in MRS culture medium at 37deg.C for 11.5 hr to end of logarithmic phase;
(4) Has strong tolerance to simulated gastrointestinal fluid.
Further, the lactobacillus helveticus OPB102 has a function of relieving constipation and diarrhea.
Use of lactobacillus helveticus OPB102 as described above for the preparation of a general food and/or a special functional food and/or a health food and/or a medicament for alleviating constipation and/or diarrhea.
Further, the food or drug has at least one of the following effects:
improving constipation, in particular shortening defecation time, increasing fecal quantity and quality and increasing small intestine propulsion rate;
alternatively, improving constipation, specifically restoring damaged colon tissue, improving villus density and increasing goblet cell number, restoring normal colon tissue, providing a better colonization environment for the flora;
or improving constipation, in particular improving intestinal flora environment, promoting beneficial bacteria such as lactobacillus and the like to reproduce, and inhibiting harmful flora from reproducing;
alternatively, the diarrhea is ameliorated, specifically the diarrhea loose stool rate, loose stool grade, and diarrhea index.
Further, the lactobacillus helveticus OPB102 is a live strain, a strain metabolite or an inactivated strain;
further, the medicament also contains a pharmaceutically acceptable carrier, which comprises: one or more of a filler, diluent, binder, disintegrant, and lubricant.
Further, the filler is one or more of microcrystalline cellulose, lactose, mannitol, starch or dextrin; the diluent is one or more of ethanol or glycerol; the disintegrating agent is one or more of sodium carboxymethyl starch, crosslinked povidone or low-substituted hydroxypropyl cellulose; the adhesive is one or more of starch paste, syrup, maltose, refined honey or liquid glucose; the lubricant is one or more of magnesium stearate, sodium stearate fumarate, talcum powder or silicon dioxide.
The application of lactobacillus helveticus OPB102 in preparing fermented food, metazoan and ferment is described.
Further, the application comprises using lactobacillus helveticus OPB102 as a fermenting microorganism, and fermenting with food material.
The invention has the advantages and positive effects that:
1. the lactobacillus helveticus OPB102 can resist the gastrointestinal environment, improve the survival rate of the gastrointestinal tract, is beneficial to the field planting of strains, and can be used for preparing functional microbial agents, foods and medicines for relieving constipation or diarrhea; the lactobacillus helveticus OPB102 can simultaneously carry out bidirectional regulation of diarrhea and constipation.
2. The lactobacillus helveticus OPB102 disclosed by the invention has the advantages that the experiment on toxic and side effects shows that the effect on mice is not obvious, the weight is not obviously changed, the death phenomenon is not generated, and obvious pathological symptoms are not generated.
3. The lactobacillus helveticus OPB102 can accelerate the defecation time of a constipation mouse, increase the number of fecal grains and the quality of the feces, and promote the small intestine to advance; at the same time, the damaged colon tissue is restored, the villus density is improved, the goblet cell number is increased, and the normal colon tissue is restored.
4. Through genome analysis of intestinal flora, the invention can effectively change the structure of intestinal flora of a constipation mouse, improve the intestinal environment, simultaneously increase the abundance of various probiotics, reduce pathogenic bacteria and effectively improve the structure of the intestinal flora by taking single strain lactobacillus helveticus OPB 102. Can reduce the loose stool rate, loose stool grade and diarrhea index of diarrhea mice.
Drawings
FIG. 1 is a colony morphology of Lactobacillus helveticus OPB102 according to the invention;
FIG. 2 is a microscopic photograph of Lactobacillus helveticus OPB102 according to the invention;
FIG. 3 is a graph showing the histological characteristics of Lactobacillus helveticus OPB102 versus constipation mice colon according to the invention; wherein, (a) H & E staining (200 x magnification); (B) AB-PAS staining (200-fold magnification);
FIG. 4 is a Venn diagram showing the change of Lactobacillus helveticus OPB102 against intestinal flora in constipation mice according to the invention;
FIG. 5 is a bar graph of the composition of Lactobacillus helveticus OPB102 versus the family horizontal colony of constipation mice intestinal flora change in accordance with the present invention;
FIG. 6 is a bar graph of the composition of Lactobacillus helveticus OPB102 according to the invention for a horizontal colony of a constipation mouse with a change in intestinal flora;
FIG. 7 is a visual circle of the change of Lactobacillus helveticus OPB102 in constipation mice intestinal flora according to the invention;
FIG. 8 is a graph of BLAST results versus tree for the Lactobacillus helveticus OPB102 genomic sequence according to the invention;
lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea is named: OPB102, class name: lactobacillus helveticus (Lactobacillus helveticus), accession number: cgmccno.26806, date of preservation: 2023, 2, 14, deposit unit: china general microbiological culture Collection center (CGMCC) of the China Committee for culture Collection of microorganisms, beijing, chaoyang area, north Chenxi Lu No. 1, 3.
Detailed Description
The invention will now be further illustrated by reference to the following examples, which are intended to be illustrative, not limiting, and are not intended to limit the scope of the invention.
The raw materials used in the invention are conventional commercial products unless otherwise specified, the methods used in the invention are conventional methods in the art unless otherwise specified, and the mass of each substance used in the invention is conventional.
Lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea is named: OPB102, class name: lactobacillus helveticus (Lactobacillus helveticus), accession number: cgmccno.26806, date of preservation: 2023, 2, 14, deposit unit: china general microbiological culture Collection center (CGMCC) of the China Committee for culture Collection of microorganisms, beijing, chaoyang area, north Chenxi Lu No. 1, 3.
Preferably, the lactobacillus helveticus OPB102 has the following biological properties:
(1) Characteristics of the cells: gram-positive, non-sporulating, non-motile bacteria;
(2) Colony characteristics: the convex part is smooth and neat;
(3) Growth characteristics: culturing in MRS culture medium at 37deg.C for 11.5 hr to end of logarithmic phase;
(4) Has strong tolerance to simulated gastrointestinal fluid.
Preferably, the lactobacillus helveticus OPB102 has the function of relieving constipation and diarrhea.
Use of lactobacillus helveticus OPB102 as described above for the preparation of a general food and/or a special functional food and/or a health food and/or a medicament for alleviating constipation and/or diarrhea.
Preferably, the food or medicament has at least one of the following effects:
improving constipation, in particular shortening defecation time, increasing fecal quantity and quality and increasing small intestine propulsion rate;
alternatively, improving constipation, specifically restoring damaged colon tissue, improving villus density and increasing goblet cell number, restoring normal colon tissue, providing a better colonization environment for the flora;
or improving constipation, in particular improving intestinal flora environment, promoting beneficial bacteria such as lactobacillus and the like to reproduce, and inhibiting harmful flora from reproducing;
alternatively, the diarrhea is ameliorated, specifically the diarrhea loose stool rate, loose stool grade, and diarrhea index.
Preferably, the lactobacillus helveticus OPB102 is a live strain, a strain metabolite or an inactivated strain;
preferably, the medicament further comprises a pharmaceutically acceptable carrier, the pharmaceutically acceptable carrier comprising: one or more of a filler, diluent, binder, disintegrant, and lubricant.
Preferably, the filler is one or more of microcrystalline cellulose, lactose, mannitol, starch or dextrin; the diluent is one or more of ethanol or glycerol; the disintegrating agent is one or more of sodium carboxymethyl starch, crosslinked povidone or low-substituted hydroxypropyl cellulose; the adhesive is one or more of starch paste, syrup, maltose, refined honey or liquid glucose; the lubricant is one or more of magnesium stearate, sodium stearate fumarate, talcum powder or silicon dioxide.
The application of lactobacillus helveticus OPB102 in preparing fermented food, metazoan and ferment is described.
Preferably, the use comprises fermenting the food material with lactobacillus helveticus OPB102 as a fermenting microorganism.
Specifically, the relevant preparation and detection steps are as follows:
example 1: screening of Lactobacillus helveticus OPB102
Isolation and screening of Lactobacillus
(1) 0.1ml of yoghurt sample (self-brewed yoghurt in the home of the herdsman in Kaisha of Xinjiang) is inoculated into a skim milk culture medium and placed in a constant temperature incubator at 37 ℃ for 48 hours. A small amount of bacterial liquid is picked by an inoculating loop, streaked and inoculated in an MRS agar culture medium plate.
(2) After 48 hours, different colonies are picked up by an inoculating loop according to the shape, size, color and the like of the colonies, and streaked separation and purification are carried out.
(3) Gram-positive bacteria and catalase-negative bacteria were retained by gram staining and catalase analysis.
(4) Obtaining thallus and colony characteristics of the final strain: gram-positive, non-sporulating, non-motile bacteria (fig. 1); has bulges and is smooth and neat (figure 2).
Molecular biological identification of Lactobacillus
(1) Single genome extraction
(A) Culturing the screened lactobacillus overnight;
(B) Using a Soxhaust bacteria genome DNA extraction kit, the product number is 1600, taking 1ml of the bacterial suspension cultured overnight, centrifuging in a 1.5ml centrifuge tube at 12000r/min for 1min, and sucking out the supernatant as much as possible;
(C) Adding 200ul of lysozyme with a final concentration of 20mg/ml into thalli, and treating for 40min at 37 ℃;
(D) Adding solution A (10 mmol/L, pH =8.0 Tris-HCl and 1mmol/L EDTA mixed solution, SDS with concentration of 0.08g/mL and NaCl with concentration of 1.5 mol/L) into the mixed solution obtained in the step (C), blowing by a pipette to fully suspend the thalli, adding 20ul rNaseA (10 mg/mL) into the suspension, fully reversing and uniformly mixing, and standing at room temperature for 30min;
(E) Adding 20ul proteinase K into the tube, mixing well, and digesting at 55deg.C for 60min; inverting the centrifuge tube for several times during digestion until the sample is clear and viscous, and proving that the sample is completely digested;
(F) Adding 200ul of solution B (mixed solution of phenol and isoamyl alcohol=25:24:1 by volume ratio) into the inquiry tube, fully inverting and uniformly mixing, if white precipitation appears, standing at 75 ℃ for 15-30 min, wherein the precipitation can disappear;
(E) Adding 200ul of absolute ethyl alcohol into a tube, fully and uniformly mixing, wherein flocculent precipitate can be generated at the moment without influencing DNA extraction, adding the solution and the flocculent precipitate into an adsorption column, and standing for 2min;
(F) Centrifuging at 12000rpm for 2min, discarding the waste liquid, and placing the adsorption column into a collecting pipe;
(G) Adding 600ul of rinsing liquid (75% ethanol) into the adsorption column, centrifuging at 12000rpm for 1min, discarding the waste liquid, placing the adsorption column into a collecting pipe, and repeating for one time;
(H) Centrifuging at 12000rpm for 2min, and placing the adsorption column in a 50 deg.C incubator for several minutes;
(I) Placing the adsorption column into a sterilized 1.5ml centrifuge tube, suspending and dripping 100ul of the washing liquid preheated by a water bath at 65 ℃ into the center of the adsorption film, standing for 5min at room temperature, and centrifuging for 1min at 12000 rpm;
(J) And adding the eluent obtained by centrifugation into an adsorption column, standing for 2min at room temperature, and centrifuging for 2min at 12000rpm to obtain the genome DNA of the lactobacillus.
The 16S rDNA gene sequence of the Lactobacillus helveticus OPB102 is as follows:
GAATGGCGGGCGTGCTAATACATGCAAGTCGAGCGAGCAGAACCAGCAGATTTACTTCGGTAATGACGCTGGGGACGCGAGCGGCGGATGGGTGAGTAACACGTGGGGAACCTGCCCCATAGTCTGGGATACCACTTGGAAACAGGTGCTAATACCGGATAAGAAAGCAGATCGCATGATCAGCTTATAAAAGGCGGCGTAAGCTGTCGCTATGGGATGGCCCCGCGGTGCATTAGCTAGTTGGTAAGGTAACGGCTTACCAAGGCAATGATGCATAGCCGAGTTGAGAGACTGATCGGCCACATTGGGACTGAGACACGGCCCAAACTCCTACGGGAGGCAGCAGTAGGGAATCTTCCACAATGGACGCAAGTCTGATGGAGCAACGCCGCGTGAGTGAAGAAGGTTTTCGGATCGTAAAGCTCTGTTGTTGGTGAAGAAGGATAGAGGTAGTAACTGGCCTTTATTTGACGGTAATCAACCAGAAAGTCACGGCTAACTACGTGCCAGCAGCCGCGGTAATACGTAGGTGGCAAGCGTTGTCCGGATTTATTGGGCGTAAAGCGAGCGCAGGCGGAAGAATAAGTCTGATGTGAAAGCCCTCGGCTTAACCGAGGAACTGCATCGGAAACTGTTTTTCTTGAGTGCAGAAGAGGAGAGTGGAATTCCATGTGTAGCGGTGGAATGCGTAGATATATGGAAGAACACCAGTGGCGAAGGCGACTCTCTGGTCTGCAACTGACGCTGAGGCTCGAAAGCATGGGTAGCGAACAGGATTAGATACCTGGTAGTCCATGCCGTAAACGATGAGTGCTAAGTGTTGGGAGGTTTCCGCCTCTCAGTGCTGCAGCTAACGCATTAAGCACTCCGCCTGGGGAGTACGACCGCAAGGTTGAAACTCAAAGGAATTGACAGGGGCCCGCACAAGCGGTGGAGCATGTGGTTTAATTCGAAGCAACGCGAAGACCTTACCAGTCTTGACATCTAGTGCCATCCTAGAGATAGAGTTCCCTTCGGGACGCTAGACAGTGTGCATGCTGTCGTCAGCTCGTGTCGTGAGATGTTGGGTTAAGTCCCGCAACGAGCGGAAATTGCTGCTTAGACGGCTCCTTCCCGAAGGTTAGGCCACCGGCTTTGGGCATTGCAGACTTCCATGGTGTGACGGGCGGTGTGTACAAGGCCCGGGAACGTATTCACCGCGGCGTTCTGATCCGCGATTACTAGCGATTCCAGCTTCGTGCAGTCGAGTTGCAGACTGCAGTCCGAACTGAGAACAGCTTTCAGAGATTCGCTTGCCTTCGCAGGCTCGCTTCTCGTTGTACTGTCCATTGTAGCACGTGTGTAGCCCAGGTCATAAGGGGCATGATGACTTGACGTCATCCCCACCTTCCTCCGGTTTATCACCGGCAGTCTCATTAGAGTGCCCAACTTAATGCTGGCAACTAATAACAAGGGTTGCGCTCGTTGCGGGACTTAACCCAACATCTCACGACACGAGCTGACGACAGCCATGCACCACCTGTCTTAGCGTCCCCGAAGGGAACTCCTAATCTCTTAGGATGGCACTAGATGTCAAGACCTGGTAAGGTTCTTCGCGTTGCTTCGAATTAAACCACATGCTCCACCGCTTGTGCGGGCCCCCGTCAATTCCTTTGAGTTTCAACCTTGCGGTCGTACTCCCCAGGCGGAGTGCTTAATGCGTTAGCTGCAGCACTGAGAGGCGGAAACCTCCCAACACTTAGCACTCATCGTTTACGGCATGGACTACCAGGGTATCTAATCCTGTTCGCTACCCATGCTTTCGAGCCTCAGCGTCAGTTGCAGACCAGAGAGTCGCCTTCGCCACTGGTGTTCTTCCATATATCTACGCATTCCACCGCTACACATGGAATTCCACTCTCCTCTTCTGCACTCAAGAAAAACAGTTTCCGATGCAGTTCCTCGGTTAAGCCGAGGGCTTTCACATCAGACTTATTCTTCCGCCTGCGCTCGCTTTACGCCCAATAAATCCGGGACACGCTTGCCACCTACGTATTACCGCGGCTGCTGGCACGTAGTTAGCCGTGACTTTCTGGTTGATTACCGTCAAATAAAGGCAGTACTACTCTATCGTTCTCACACACAGAGCTTTACGATCCGAAACTTCTCACTCACGCGCG。
（2）16S r DNA PCR
bacterial 16S r DNA 50. Mu. LPCR reaction System: 24 mu L of Taq enzyme; dd H2O: 18. Mu.L; 27F, 2. Mu.L; 1492R, 2. Mu.L; and (3) a template: 4. Mu.L.
PCR conditions: 94℃for 5min (i.e.step 1); 94℃for 30s (i.e.step 2); 55℃for 30s (i.e.step 3); 30s at 72 ℃ (i.e. step 4); step2 to step4 (30×); and at 72℃for 5min.
Preparing 1% agarose gel, mixing the PCR product with 10000×loading buffer, loading 2 μl, running at 120V for 30min, and performing gel imaging;
the PCR products were sent to a professional sequencing company, and the sequencing results obtained were compared with searches and similarities performed in Gen Bank using BLAST (see FIG. 8), and identified as strains of Lactobacillus helveticus, and stored at-80 ℃.
Example 2: lactobacillus helveticus OPB102 has good tolerance to simulated gastrointestinal fluids
In vitro simulated gastric fluid: the pH value of the 0.1mol/L potassium phosphate buffer solution is respectively adjusted to 2.5, pepsin (10 g/L) is added, and the mixture is uniformly mixed and then passes through a 0.22 mu m sterile film, so that simulated gastric juice can be obtained.
In vitro simulated intestinal juice: the pH value of the 0.1mol/L potassium phosphate buffer solution is regulated to 6.8, pancreatin (10 g/L) and pig bile salt (3 g/L) are added, and the mixture is uniformly mixed and then passes through a 0.22 mu m sterile film, thus obtaining the simulated intestinal juice.
Gastric digestion was simulated:
taking culture solution of 3.0 mL Lactobacillus helveticus OPB102, centrifuging for 3min, collecting mycelium, placing in 3.0 mL of pH 2.5 simulated gastric juice environment, and simulating gastric juice digestion under anaerobic condition of 37deg.C and 90 r/min. Samples were taken at 0, 1.0, 2.0, 3.0 and h, viable bacteria were counted using a dilution-coated plate count method, and survival rates were calculated.
Intestinal digestion was simulated:
centrifuging culture solution of Lactobacillus helveticus OPB102 at 3.0 mL for 3min, collecting mycelium, and placing in 3.0 mL simulated intestinal juice environment with pH value of 6.8 at 37deg.C under anaerobic condition of 120 r/min to simulate intestinal juice digestion. Samples were taken at 0, 2.0, 4.0, 6.0, 8.0 and h, viable bacteria were counted using a dilution-coated plate count method, and survival rates were calculated.
The survival rate (%) was calculated as the ratio of the number of viable bacteria at the time of sampling to the number of viable bacteria at the time of 0h in the culture solution, expressed as%. The experimental results are shown in tables 1 and 2, and the results show that the lactobacillus helveticus OPB102 has better tolerance to artificial simulated gastrointestinal fluid.
Example 3: lactobacillus helveticus OPB102 has no toxic and side effects on SFP-class Balb/c mice
Suspending Lactobacillus helveticus OPB102 in 100g/L skim milk solution to give 4.0X10-g concentration 9 CFU/mL of bacterial suspension. 40 SPF-class Balb/c mice are bred in the environment of the light irradiation time of 12 h/12 h, wherein the male body mass is 18-20 g, the temperature is 20-25 ℃, the relative humidity is 50+/-5%. The group was divided into Lactobacillus helveticus OPB102 and control group. The group of Lactobacillus helveticus OPB102 was given 0.3mL of this concentrated bacterial suspension once daily for gavage, and the control group was gavaged with the same volume of 100g/L skim milk solution without Lactobacillus helveticus OPB102, and the death and weight were recorded for one week.
The results of these tests are shown in Table 3. These results indicate that the feeding concentration is 1X 10 9 CFU/Lactobacillus helveticus OPB102 has no obvious effect on mice, no obvious change of body weight and no death phenomenon. The appearance of the mice has no obvious pathological symptoms.
Example 4: measurement of the time to stool, the number of stool particles and the stool quality of constipation mice by lactobacillus helveticus OPB 102.
30 SPF-class Balb/c mice are bred in the environment of the light irradiation time of 12 h/12 h, wherein the male body mass is 18-20 g, the temperature is 20-25 ℃, the relative humidity is 50+/-5%. 30 mice were randomly divided into 3 groups of 10 mice each, and the groups were a blank control group, a model control group and an experimental group. Model control group and experimental group mice were perfused with loperamide hydrochloride (5 mg/(kg·bw)), and were continuously perfused with stomach for 7d each time in the morning and evening, to construct constipation models. The blank group was filled with an equal amount of distilled water. After successful modeling, mice in the blank control group and the model control group are respectively filled with 0.3mL of distilled water, mice in the experiment group are filled with 0.3mL of gastric probiotic suspension (1.0 g/(kg.bw), the mice are fed normally during the test once a day, and are continuously fed for 14 days, the mice are fed in a single cage, filter paper is filled under the cage, and the defecation amount of the mice in 6 hours is recorded after 14 days.
Example 5: effect of Lactobacillus helveticus OPB102 on constipation mice intestinal ink propulsion experiments
30 SPF-class Balb/c mice are bred in the environment of the light irradiation time of 12 h/12 h, wherein the male body mass is 18-20 g, the temperature is 20-25 ℃, the relative humidity is 50+/-5%. The animals were randomly divided into 3 groups of 10 animals each, and the three groups were a blank control group, a model control group, and a probiotic experimental group. Model control group and experimental group mice were perfused with loperamide hydrochloride (5 mg/(kg·bw)), and were continuously perfused with stomach for 7d each time in the morning and evening, to construct constipation models. The blank control group and the model control group were respectively filled with 0.3mL of distilled water, 0.3mL of probiotic suspension (1.0 g/(kg. Bw), fed normally once a day during the test, fed continuously for 14 days, fasted for 24 hours after 14 days, the blank control group was given an equal volume of distilled water, after molding for 30 minutes, each group was filled with ink (0.1 mL/10 g), the mice were sacrificed after 20 minutes by cervical vertebra removal, the small intestine was opened, the upper end to the pylorus was cut, the lower end to the intestinal canal of the ileocecum, gently pulled into a straight line, the ink advancing length and the total length of the small intestine were measured, and the ink advancing rate was calculated, and the test results are shown in Table 5.
Ink advance (%) = (ink advance length/total length of small intestine) ×100%
After each group of test mice was continuously perfused with the corresponding probiotic suspension for 7 and 14 days, the molding was successful. And the ink propulsion rate test of the intestinal tracts of the mice is carried out, and the ink propulsion rate of the intestinal tracts of the mice in each group is shown in table 5. As can be seen from Table 5, the ink push rate of the model group was significantly lower than that of the blank group, indicating that the constipation model was established in the mice. Compared with the model group, the small intestine ink propulsion rate of the probiotics group is higher, the difference is obvious, the probiotics group can promote small intestine peristalsis, meanwhile, the small intestine propulsion rate of 14 days is higher than the small intestine propulsion rate of 7 days, and the probiotics group is more beneficial to improving defecation conditions after long-term administration.
Example 6: effect of Lactobacillus helveticus OPB102 on pathological changes of colon tissue of model mice
For example 5, clean colon tissue samples were cut with sterile surgical scissors 2 cm, placed in 10% formalin solution for overnight fixation, and then prepared into paraffin sections of colon tissue. Sections were stained with hematoxylin-eosin (HE staining), alisxin blue-periodate schiff stain (AB-PAS staining). The colon histopathological changes were observed under an optical microscope and stored by photographing (fig. 3).
As shown in fig. 3, the normal colon tissue morphology is normal, the intestinal glands of the tissue mucous membrane layer are orderly arranged, the structure is clear, and the villi are compact and orderly. In contrast, the colon muscle layer of the model group was thinned, the villi was broken loose, the submucosa was damaged, and the intestinal gland was disordered. The colon muscle layer of the probiotics group is thickened, the intestinal gland arrangement is close to the normal level, the damage of submucosa is reduced, the tissue morphology is improved compared with that of the model group, and the probiotics group has a certain protection effect on intestinal tracts. By observing the ratio of the villus length to the crypt depth of panel A in FIG. 3, the model group has the shortest villus, and the villus length after feeding with Lactobacillus helveticus OPB102 is recovered, thus showing that Lactobacillus helveticus OPB102 can improve the colon villus length to some extent.
After AB-PAS staining, a large number of aligned goblet cells were observed in the normal group, whereas fewer goblet cells were observed in the model group. The lactobacillus helveticus OPB102 can restore the ordered arrangement of the goblet cells, increase the number of the goblet cells, restore the damaged colon tissues, increase the number of the goblet cells, restore the normal level state of the colon tissues, is more beneficial to the colonial growth and improves constipation.
Example 7: effect of Lactobacillus helveticus OPB102 on constipation mice intestinal flora
30 SPF-class Balb/c mice are bred in the environment of the light irradiation time of 12 h/12 h, wherein the male body mass is 18-20 g, the temperature is 20-25 ℃, the relative humidity is 50+/-5%. The random number of the bacteria was 4 groups, each group was 10, and the bacteria were a blank group (NC), a model group (MC), a probiotic group (YS) (Lactobacillus helveticus OPB102, 50 hundred million CFU/g) and a mixed bacteria positive group (YX) (Lactobacillus plantarum: lactobacillus bifidus: bifidobacterium bifidus: lactobacillus acidophilus: lactobacillus paracasei mixed bacteria, the mass ratio of which was 1:1:1:1:1, and the bacteria were commercially available, 50 CFU/g). The model group, the probiotic group and the mixed bacteria positive group mice were perfused with loperamide hydrochloride (5 mg/(kg.bw)), and were continuously perfused with stomach for 7d each time in the morning and evening, to construct a constipation model. The blank group and the model control group were respectively filled with 0.3mL of distilled water, the probiotic group and the mixed bacteria positive group were filled with 0.3mL of probiotic suspension (1.0 g/(kg. Bw) of the conversion amount was dissolved in 0.3 mL), and the test period was normally fed once a day, continuously fed for 14 days, and fasted for 24 hours after 14 days. The colon content of the mice is collected in a sterile freezing tube, the operation is rapid, the pollution is avoided as much as possible in the process, all samples are immediately placed in liquid nitrogen after being collected, and the dry ice is stored and transferred to Shanghai Meiji biological medicine science and technology Co for sequencing after the sampling is finished. The PCR reaction parameters of the formal test are as follows: (a) denaturation at 95℃for 3 min; (b) Cycle number (denaturation at 95℃30 s; annealing temperature 30 s; extension at 72℃45 s); (c) extending at 72 ℃ for 10 min, and preserving heat at 10 ℃. The PCR products were detected using 2% agarose gel electrophoresis. Sequences were clustered into Operational Taxonomies (OTUs) using the USEARCH11 usperse algorithm (version 7.0.1090 http:// drive5.Com/uparse /), with a similarity of 97%. Samples are further analyzed based on the minimum number of sample sequences extracted. Data analysis was performed using IBM SPSS Statistics software and GraphPad Prism 8.0.2 software.
1. Analysis of sample species composition
The Venn diagram can be used for counting the number of common and unique species (such as OTU) in different groups, and can intuitively represent the species (such as OTU) composition uniqueness and overlapping condition of colon content samples. We analyzed the number of OTUs in four groups of mice (blank, model, positive control, lactobacillus helveticus) and plotted Venn's chart, see fig. 4. The figure shows that there are unique OUT among different samples, 551 OUT in total, and 407 OUT overlaps among blank group, model group, positive control group and lactobacillus helveticus group. The blank (NC), model (MC), positive control (YX), lactobacillus helveticus (YS) had 13, 10, 5 and 3 unique OUT, respectively. The results showed that the number of intestinal flora OTUs increased after administration in the positive control group, the lactobacillus helveticus group, and that the number of OTUs in both the YS group and the YX group was higher than that in the MC group, and more similar to that in the NC group.
2. Colony composition analysis
According to the OTU classification analysis result, species composition conditions among different groups can be known. To further more intuitively present the different dominant species of each group at a certain taxonomic level (e.g. domain, kingdom, phylum, class, order, family, genus, species, OTU, etc.), and to further elucidate the effect of lactobacillus helveticus on the composition of the intestinal flora, we analyzed the composition of the intestinal flora at the family and genus level, and the results are shown in fig. 5 and 6. As can be seen from fig. 5, lactobacillaceae, muribaculaceae and Lachnospiraceae are three dominant bacterial groups in all groups at the family level. The percentage of Muribaculaceae, lactobacilliaceae and Lachnospiraceae in the whole flora was approximately 70%, and the three beneficial bacteria in the model group were relatively reduced by only 60% compared to the blank group. After lactobacillus helveticus treatment, the total number of Lactobacillaceae, muribaculaceae and Lachnospiraceae all tended to rise, especially the rising of Lactobacillaceae and Muribaculaceae was most obvious. Furthermore, eggerthellac is the most pathogenic bacteria in the current study that promote enteritis, and is highest in the model group, and the blank group, the positive control group and the probiotic group are relatively low. Prevoltellaceae is a commensal intestinal bacterium involved in intestinal mucosal mucus formation and SCFA production, and is more abundant in the NC, YX and YS groups than in the MC group. The increase in Prevotellaceae contributes to the formation of intestinal mucosa. This is consistent with the results of the colon section staining study above.
As can be seen from fig. 6, the MC group lactic acid bacteria content is significantly reduced at the genus level compared to the blank group, which may be related to the side effects of loperamide hydrochloride. The lactic acid bacteria content of the YX and YS groups is obviously increased, and the levels of the YX and YS groups are close. The results show that lactobacillus helveticus increases the content of lactobacillus.
3. Relationship between sample and species
Differences in bacterial genus between groups were investigated by analyzing the relationship between samples and species. As shown in FIG. 7, the distribution ratio of each dominant species in different groups is shown by a visual circle chart. Each group of flora consists essentially of Lactobacillus, norank_f __ Muribaculaceae, norank_f __ lachnospirace, unclassified_f __ lachnospirace and bacterioides. Lactobacillus accounts for 32% of the blank and 19% of the model. Compared with the model group, the Lactobacillus content of each treatment group is obviously increased, and the Lactobacillus content after YX and YS treatment is 25% and 24%, respectively. The improvement in the YX and YS treatment groups is particularly significant. Lactobacillus inhibits intestinal pathogenic bacteria by enhancing mucosal barrier, stimulates goblet cells of intestinal epithelium, and generates a large amount of mucus-inhibiting pathogenic bacteria to invade intestinal epithelium. In addition, lactobacillus tightly binds to the intestinal epithelium and increases the intercellular integrity of the tight junction at the tip to maintain intestinal permeability. Norank_f __ Muribaculaceae represents 24% and 22% respectively in the blank and model groups. After YX and YS treatments, the norank_f __ Muribaculaceae accounted for 25% and 29%, respectively. The proportion of norank_f __ Muribaculaceae in the model group was decreased compared to the blank group, while the proportion of YX and YS treatments was increased, approaching the blank group.
Example 8: effect of Lactobacillus helveticus OPB102 on the rare rate, rare grade and diarrhea index of diarrhea mice
40 SPF-class Balb/c mice are bred in the environment of the light irradiation time of 12 h/12 h, wherein the male body mass is 18-20 g, the temperature is 20-25 ℃, the relative humidity is 50+/-5%. The cells were randomly divided into 4 groups of 10 cells, and the cells were selected from a blank control group, a model group, a probiotic test group and a positive control group (Yimengding). The mice in the blank control group were given 10 mL/kg of the vehicle (3.2% Tween-80 aqueous solution), the mice in the model group were given 10 mL/kg of the vehicle (3.2% Tween-80 aqueous solution), the mice in the probiotic experimental group were given 1.0g/kg of the probiotic (3.2% Tween-80 aqueous solution as the vehicle), and the mice in the positive control group were given 10 mg/kg of Yimengting (3.2% Tween-80 solution as the vehicle) 1 time/d for 7 d. After the last administration of 1 h, the mice in the model group, the probiotics experiment group and the positive control group are subjected to gastric administration, castor oil 10 mL/kg is administered, the mice are immediately placed in a mouse cage, the normal stool number, the loose stool number and the loose stool level of the mice at each interval of 1 h in 5h are observed and recorded, and the diarrhea index is calculated according to the loose stool rate and the loose stool level. The calculation formula of the loose stool rate is as follows: rare stool rate = number of rare stools discharged in a certain time/total number of bowel stools diarrhea index calculation formula: diarrhea index = runny bowel rate x runny bowel grade. Compared with the control group, the probiotic bacteria and the diarrhea index of the mice with diarrhea after the easy Mongolian group are acted are reduced. Meanwhile, the diarrhea index of the probiotics group is slightly lower than that of the easy Mongolian group. As can be seen from table 6, the probiotic preparation can regulate intestinal flora, effectively improve castor oil-induced diarrhea in mice, and has an antidiarrheal effect similar to that of a positive control easy-to-stop group.
Comparative example 1:
the specific embodiment is the same as in example 5, except that Lactobacillus helveticus OPB102 is replaced by Lactobacillus helveticus LH-90 (commercially available strain of Jia Yi in the middle, and the 14-day propulsion rate of the small intestine ink of the constipation mice is measured, and the result shows that the propulsion rate of the Lactobacillus helveticus LH-90 to the small intestine ink of the constipation mice is 30.32+/-0.66%, which is obviously lower than the propulsion rate of the probiotics group 42.43 +/-1.74%. It can be seen that lactobacillus helveticus OPB102 has a significant effect on constipation mice intestinal thrust. Compared with the propulsion rate of the Lactobacillus helveticus LH-90, the Lactobacillus helveticus OPB102 relieves constipation and promotes small intestine peristalsis, which is obviously superior to the Lactobacillus helveticus LH-90.
Comparative example 2
The specific embodiment is the same as example 8, except that Lactobacillus helveticus OPB102 is replaced by Lactobacillus helveticus LH-90 (commercially available strain of Jia Yi in the middle, and the loose stool rate, loose stool grade and diarrhea index of diarrhea mice are measured, and the result shows that the Lactobacillus helveticus LH-90 has no alleviation effect on the loose stool rate, loose stool grade and diarrhea index of diarrhea mice compared with the diarrhea model group (0.82+ -0.07, 1.23+ -0.21, 1.01+ -0.13) of the diarrhea mice, wherein the loose stool rate, loose stool grade and diarrhea index of diarrhea mice are respectively 0.78+ -0.12, 1.21+ -0.09, 1.0+ -0.03.
Although embodiments of the present invention have been disclosed for illustrative purposes, those skilled in the art will appreciate that: various substitutions, changes and modifications are possible without departing from the spirit and scope of the invention and the appended claims, and therefore the scope of the invention is not limited to the disclosure of the embodiments.
Claims (10)
1. A lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea, characterized in that: the classification names are: lactobacillus helveticus（Lactobacillus helveticus）The preservation number is: CGMCC No.26806, date of preservation: 2023, 2, 14, deposit unit: china general microbiological culture Collection center, north Chen Xi Lu No. 1, 3, the Korean region of Beijing, and the China general microbiological culture Collection center.
2. Lactobacillus helveticus OPB102 according to claim 1, characterized in that: the lactobacillus helveticus OPB102 has the following biological properties:
(1) Characteristics of the cells: gram-positive, non-sporulating, non-motile bacteria;
(2) Colony characteristics: the convex part is smooth and neat;
(3) Growth characteristics: culturing in MRS culture medium at 37deg.C for 11.5 hr to end of logarithmic phase;
(4) Has strong tolerance to simulated gastrointestinal fluid.
3. Lactobacillus helveticus OPB102 according to claim 1 or 2, characterized in that: the lactobacillus helveticus OPB102 has the function of relieving constipation and diarrhea.
4. Use of lactobacillus helveticus OPB102 according to any of the claims 1 to 3 for the preparation of a general food and/or a special function food and/or a health food and/or a medicament for alleviating constipation and/or diarrhea.
5. The use according to claim 4, characterized in that: the food or medicament has at least one of the following effects:
improving constipation, in particular shortening defecation time, increasing fecal quantity and quality and increasing small intestine propulsion rate;
alternatively, improving constipation, specifically restoring damaged colon tissue, improving villus density and increasing goblet cell number, restoring normal colon tissue, providing a better colonization environment for the flora;
or, improving constipation, in particular improving intestinal flora environment, promoting beneficial bacteria reproduction and inhibiting harmful bacteria reproduction;
alternatively, the diarrhea is ameliorated, specifically the diarrhea loose stool rate, loose stool grade, and diarrhea index.
6. The use according to claim 4, characterized in that: the lactobacillus helveticus OPB102 is a live strain, a strain metabolite or an inactivated strain.
7. The use according to claim 4, characterized in that: the medicament further comprises a pharmaceutically acceptable carrier, which comprises: one or more of a filler, diluent, binder, disintegrant, and lubricant.
8. The use according to claim 7, characterized in that: the filler is one or more of microcrystalline cellulose, lactose, mannitol, starch or dextrin; the diluent is one or more of ethanol or glycerol; the disintegrating agent is one or more of sodium carboxymethyl starch, crosslinked povidone or low-substituted hydroxypropyl cellulose; the adhesive is one or more of starch paste, syrup, maltose, refined honey or liquid glucose; the lubricant is one or more of magnesium stearate, sodium stearate fumarate, talcum powder or silicon dioxide.
9. Use of lactobacillus helveticus OPB102 according to any of the claims 1 to 3 for the preparation of fermented food products, metazoan, ferments.
10. The use according to claim 9, characterized in that: the application comprises using Lactobacillus helveticus OPB102 as a fermentation microorganism, and fermenting with food material.
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311051207.7A CN116790448B (en) | 2023-08-21 | 2023-08-21 | Lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202311051207.7A CN116790448B (en) | 2023-08-21 | 2023-08-21 | Lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN116790448A true CN116790448A (en) | 2023-09-22 |
| CN116790448B CN116790448B (en) | 2024-01-12 |
Family
ID=88044000
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202311051207.7A Active CN116790448B (en) | 2023-08-21 | 2023-08-21 | Lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN116790448B (en) |
Cited By (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117106674A (en) * | 2023-10-23 | 2023-11-24 | 新益(天津)生物科技有限责任公司 | Lactobacillus plantarum OPB15 capable of reducing uric acid and repairing microecological balance of female private parts and application |
| CN117100773A (en) * | 2023-10-25 | 2023-11-24 | 山东威曼宠物食品有限公司 | Lactobacillus plantarum King07 metazoan preparation and application thereof |
Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20000054037A (en) * | 2000-05-19 | 2000-09-05 | 윤병대 | Novel strain lactobacillus helveticus brd-001 having thermal and acid resistance and a cultivating method with high concentration |
| CN106722206A (en) * | 2017-01-09 | 2017-05-31 | 广州能靓生物技术有限公司 | A kind of composition for alleviating alimentary canal malaise symptoms and/or emotional stress and products thereof |
| CN112210516A (en) * | 2020-10-19 | 2021-01-12 | 河北一然生物科技有限公司 | Lactobacillus helveticus L1258 with intestinal tract regulation function and composition thereof |
| CN112877233A (en) * | 2021-01-23 | 2021-06-01 | 江苏斯卡露生物科技有限公司 | Lactobacillus helveticus strain and application thereof |
| WO2022196488A1 (en) * | 2021-03-15 | 2022-09-22 | 森永乳業株式会社 | Composition for improving qol |
-
2023
- 2023-08-21 CN CN202311051207.7A patent/CN116790448B/en active Active
Patent Citations (5)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| KR20000054037A (en) * | 2000-05-19 | 2000-09-05 | 윤병대 | Novel strain lactobacillus helveticus brd-001 having thermal and acid resistance and a cultivating method with high concentration |
| CN106722206A (en) * | 2017-01-09 | 2017-05-31 | 广州能靓生物技术有限公司 | A kind of composition for alleviating alimentary canal malaise symptoms and/or emotional stress and products thereof |
| CN112210516A (en) * | 2020-10-19 | 2021-01-12 | 河北一然生物科技有限公司 | Lactobacillus helveticus L1258 with intestinal tract regulation function and composition thereof |
| CN112877233A (en) * | 2021-01-23 | 2021-06-01 | 江苏斯卡露生物科技有限公司 | Lactobacillus helveticus strain and application thereof |
| WO2022196488A1 (en) * | 2021-03-15 | 2022-09-22 | 森永乳業株式会社 | Composition for improving qol |
Cited By (4)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN117106674A (en) * | 2023-10-23 | 2023-11-24 | 新益(天津)生物科技有限责任公司 | Lactobacillus plantarum OPB15 capable of reducing uric acid and repairing microecological balance of female private parts and application |
| CN117106674B (en) * | 2023-10-23 | 2024-03-08 | 新益(天津)生物科技有限责任公司 | Lactobacillus plantarum OPB15 capable of reducing uric acid and repairing microecological balance of female private parts and application |
| CN117100773A (en) * | 2023-10-25 | 2023-11-24 | 山东威曼宠物食品有限公司 | Lactobacillus plantarum King07 metazoan preparation and application thereof |
| CN117100773B (en) * | 2023-10-25 | 2024-02-09 | 山东威曼宠物食品有限公司 | Lactobacillus plantarum King07 metazoan preparation and application thereof |
Also Published As
| Publication number | Publication date |
|---|---|
| CN116790448B (en) | 2024-01-12 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN116790448B (en) | Lactobacillus helveticus OPB102 capable of relieving constipation and diarrhea and application thereof | |
| CN110604749B (en) | Bifidobacterium animalis A12 and its application in controlling diabetes or hyperlipidemia, especially weight gain or obesity | |
| CN109628346B (en) | Lactobacillus fermentum CQPC04 and application thereof in preparing fermented food | |
| EP4053262A1 (en) | Lactobacillus casei producing short-chain fatty acids, cultivation method therefor and application thereof | |
| WO2022206300A1 (en) | Composition capable of facilitating defecation and use thereof | |
| CN113122467B (en) | Lactobacillus paracasei and composition thereof | |
| CN116555076B (en) | Bifidobacterium longum subspecies longum MY1 and application thereof in preparation of food and medicine for relaxing bowels and protecting intestines | |
| CN116286551B (en) | Application of bifidobacterium longum subspecies infantis in regulating in-vivo fat metabolism, shaping, reducing fat and improving obesity | |
| CN112218646A (en) | Composition and application thereof | |
| CN114774315B (en) | Application of lactobacillus rhamnosus strain LRa05 in preparation of immunity enhancing product and/or eczema relieving product | |
| CN114657084B (en) | Bifidobacterium longum for relieving ulcerative colitis and application thereof | |
| CN113249280A (en) | Streptococcus thermophilus STN26, bacterium powder and application in uric acid reducing product | |
| JP4248985B2 (en) | Lactobacillus fermentum, a preparation for preventing or treating obesity or diabetes, and a food composition comprising the same and a carrier | |
| CN116814510B (en) | Lactobacillus rhamnosus OPB41 capable of preventing or improving Alzheimer's disease and application thereof | |
| CN109207389B (en) | Thrombolytic lipid-lowering probiotic compound bacteria traditional Chinese medicine oral liquid and preparation method thereof | |
| CN117004503B (en) | Saliva combined lactobacillus MB1 and application thereof in preparation of food and medicine for assisting sleep and regulating intestines and stomach | |
| CN113512514A (en) | Lactococcus lactis with depression improving effect and application thereof | |
| CN116555075B (en) | Lactobacillus plantarum JF1 and application thereof in preparation of anti-aging food and drug | |
| CN117264814A (en) | Lactobacillus rhamnosus with effects of preventing and treating digestive tract diseases | |
| CN116445356A (en) | Bifidobacterium animalis subspecies BA67 for regulating intestinal flora and enhancing immunity and application thereof | |
| CN107929329B (en) | Thrombolytic lipid-lowering probiotic composite bacteria traditional Chinese medicine granule and preparation method thereof | |
| CN114933992A (en) | Bifidobacterium longum and application of compound preparation thereof in relieving ulcerative colitis | |
| CN114836349A (en) | Lactobacillus acidophilus LA16 for antagonizing helicobacter pylori and application thereof | |
| CN116496938B (en) | Lactobacillus acidophilus MY2 for producing hyaluronic acid and application thereof in preparation of anti-aging and whitening food and medicines | |
| CN116622593B (en) | Lactobacillus paracasei for fermentation and fermentation process for preparing wind-resistant acid-discharging ferment by same |
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 |