CN117986337B - 一种提高免疫力的益生菌组合物以及在治疗疾病中的应用 - Google Patents
一种提高免疫力的益生菌组合物以及在治疗疾病中的应用 Download PDFInfo
- Publication number
- CN117986337B CN117986337B CN202410149675.6A CN202410149675A CN117986337B CN 117986337 B CN117986337 B CN 117986337B CN 202410149675 A CN202410149675 A CN 202410149675A CN 117986337 B CN117986337 B CN 117986337B
- Authority
- CN
- China
- Prior art keywords
- inflammatory
- probiotic
- lactobacillus
- probiotic composition
- 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.)
- Active
Links
- 239000006041 probiotic Substances 0.000 title claims abstract description 68
- 235000018291 probiotics Nutrition 0.000 title claims abstract description 68
- 230000000529 probiotic effect Effects 0.000 title claims abstract description 36
- 239000000203 mixture Substances 0.000 title claims abstract description 27
- 230000036039 immunity Effects 0.000 title claims abstract description 13
- 201000010099 disease Diseases 0.000 title abstract description 6
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 title abstract description 6
- 108090000765 processed proteins & peptides Proteins 0.000 claims abstract description 59
- 230000003110 anti-inflammatory effect Effects 0.000 claims abstract description 27
- 241000186606 Lactobacillus gasseri Species 0.000 claims abstract description 9
- 229920001184 polypeptide Polymers 0.000 claims description 35
- 102000004196 processed proteins & peptides Human genes 0.000 claims description 35
- 208000022559 Inflammatory bowel disease Diseases 0.000 claims description 21
- 239000003814 drug Substances 0.000 claims description 8
- 241000218588 Lactobacillus rhamnosus Species 0.000 claims description 5
- 240000002605 Lactobacillus helveticus Species 0.000 claims description 4
- 235000013967 Lactobacillus helveticus Nutrition 0.000 claims description 4
- 230000002708 enhancing effect Effects 0.000 claims description 4
- 229940054346 lactobacillus helveticus Drugs 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 238000004519 manufacturing process Methods 0.000 claims description 3
- 125000003275 alpha amino acid group Chemical group 0.000 claims 4
- 241000186660 Lactobacillus Species 0.000 abstract description 16
- 230000000694 effects Effects 0.000 abstract description 16
- 229940039696 lactobacillus Drugs 0.000 abstract description 16
- 241000699670 Mus sp. Species 0.000 abstract description 13
- 206010009900 Colitis ulcerative Diseases 0.000 abstract description 10
- 201000006704 Ulcerative Colitis Diseases 0.000 abstract description 9
- 210000004027 cell Anatomy 0.000 description 11
- 206010061218 Inflammation Diseases 0.000 description 10
- 230000004054 inflammatory process Effects 0.000 description 10
- -1 oleyl alcohols Chemical class 0.000 description 10
- 230000000968 intestinal effect Effects 0.000 description 8
- 208000002551 irritable bowel syndrome Diseases 0.000 description 7
- GOZMBJCYMQQACI-UHFFFAOYSA-N 6,7-dimethyl-3-[[methyl-[2-[methyl-[[1-[3-(trifluoromethyl)phenyl]indol-3-yl]methyl]amino]ethyl]amino]methyl]chromen-4-one;dihydrochloride Chemical compound Cl.Cl.C=1OC2=CC(C)=C(C)C=C2C(=O)C=1CN(C)CCN(C)CC(C1=CC=CC=C11)=CN1C1=CC=CC(C(F)(F)F)=C1 GOZMBJCYMQQACI-UHFFFAOYSA-N 0.000 description 6
- 210000001035 gastrointestinal tract Anatomy 0.000 description 6
- KBOPZPXVLCULAV-UHFFFAOYSA-N mesalamine Chemical group NC1=CC=C(O)C(C(O)=O)=C1 KBOPZPXVLCULAV-UHFFFAOYSA-N 0.000 description 6
- 239000013642 negative control Substances 0.000 description 6
- 102000004890 Interleukin-8 Human genes 0.000 description 5
- 108090001007 Interleukin-8 Proteins 0.000 description 5
- 229920003171 Poly (ethylene oxide) Polymers 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 230000001965 increasing effect Effects 0.000 description 5
- 239000002609 medium Substances 0.000 description 5
- 230000026731 phosphorylation Effects 0.000 description 5
- 238000006366 phosphorylation reaction Methods 0.000 description 5
- 239000013641 positive control Substances 0.000 description 5
- 239000004094 surface-active agent Substances 0.000 description 5
- 208000011231 Crohn disease Diseases 0.000 description 4
- 102000043136 MAP kinase family Human genes 0.000 description 4
- 108091054455 MAP kinase family Proteins 0.000 description 4
- 108060008682 Tumor Necrosis Factor Proteins 0.000 description 4
- 102100040247 Tumor necrosis factor Human genes 0.000 description 4
- 210000001072 colon Anatomy 0.000 description 4
- 230000006378 damage Effects 0.000 description 4
- 210000002919 epithelial cell Anatomy 0.000 description 4
- 230000002757 inflammatory effect Effects 0.000 description 4
- 210000004347 intestinal mucosa Anatomy 0.000 description 4
- 230000002829 reductive effect Effects 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- LZZYPRNAOMGNLH-UHFFFAOYSA-M Cetrimonium bromide Chemical compound [Br-].CCCCCCCCCCCCCCCC[N+](C)(C)C LZZYPRNAOMGNLH-UHFFFAOYSA-M 0.000 description 3
- 240000001046 Lactobacillus acidophilus Species 0.000 description 3
- 235000013956 Lactobacillus acidophilus Nutrition 0.000 description 3
- 230000004913 activation Effects 0.000 description 3
- 244000052616 bacterial pathogen Species 0.000 description 3
- 230000007547 defect Effects 0.000 description 3
- 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 3
- 229940079593 drug Drugs 0.000 description 3
- 238000011534 incubation Methods 0.000 description 3
- 229940039695 lactobacillus acidophilus Drugs 0.000 description 3
- 230000003902 lesion Effects 0.000 description 3
- 229960004963 mesalazine Drugs 0.000 description 3
- 230000009689 neuronal regeneration Effects 0.000 description 3
- 239000002953 phosphate buffered saline Substances 0.000 description 3
- 238000011160 research Methods 0.000 description 3
- 230000019491 signal transduction Effects 0.000 description 3
- 230000001225 therapeutic effect Effects 0.000 description 3
- 210000001519 tissue Anatomy 0.000 description 3
- ULQISTXYYBZJSJ-UHFFFAOYSA-N 12-hydroxyoctadecanoic acid Chemical compound CCCCCCC(O)CCCCCCCCCCC(O)=O ULQISTXYYBZJSJ-UHFFFAOYSA-N 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 2
- 241000193830 Bacillus <bacterium> Species 0.000 description 2
- 241000186000 Bifidobacterium Species 0.000 description 2
- 102000013585 Bombesin Human genes 0.000 description 2
- 108010051479 Bombesin Proteins 0.000 description 2
- 208000004232 Enteritis Diseases 0.000 description 2
- 101000979249 Homo sapiens Neuromodulin Proteins 0.000 description 2
- 240000006024 Lactobacillus plantarum Species 0.000 description 2
- 235000013965 Lactobacillus plantarum Nutrition 0.000 description 2
- 102100023206 Neuromodulin Human genes 0.000 description 2
- DBMJMQXJHONAFJ-UHFFFAOYSA-M Sodium laurylsulphate Chemical compound [Na+].CCCCCCCCCCCCOS([O-])(=O)=O DBMJMQXJHONAFJ-UHFFFAOYSA-M 0.000 description 2
- 208000027418 Wounds and injury Diseases 0.000 description 2
- 230000006838 adverse reaction Effects 0.000 description 2
- 150000001412 amines Chemical class 0.000 description 2
- 238000010171 animal model Methods 0.000 description 2
- 230000000844 anti-bacterial effect Effects 0.000 description 2
- 239000003963 antioxidant agent Substances 0.000 description 2
- 230000003078 antioxidant effect Effects 0.000 description 2
- 230000001580 bacterial effect Effects 0.000 description 2
- DNDCVAGJPBKION-DOPDSADYSA-N bombesin Chemical compound C([C@@H](C(=O)N[C@@H](CC(C)C)C(=O)N[C@@H](CCSC)C(N)=O)NC(=O)CNC(=O)[C@@H](NC(=O)[C@H](C)NC(=O)[C@H](CC=1NC2=CC=CC=C2C=1)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H](CC(N)=O)NC(=O)CNC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCNC(N)=N)NC(=O)[C@H](CCC(N)=O)NC(=O)[C@H]1NC(=O)CC1)C(C)C)C1=CN=CN1 DNDCVAGJPBKION-DOPDSADYSA-N 0.000 description 2
- 238000004113 cell culture Methods 0.000 description 2
- 238000011161 development Methods 0.000 description 2
- 230000018109 developmental process Effects 0.000 description 2
- 235000014113 dietary fatty acids Nutrition 0.000 description 2
- 235000019329 dioctyl sodium sulphosuccinate Nutrition 0.000 description 2
- 230000009266 disease activity Effects 0.000 description 2
- SYELZBGXAIXKHU-UHFFFAOYSA-N dodecyldimethylamine N-oxide Chemical compound CCCCCCCCCCCC[N+](C)(C)[O-] SYELZBGXAIXKHU-UHFFFAOYSA-N 0.000 description 2
- 210000000981 epithelium Anatomy 0.000 description 2
- 229930195729 fatty acid Natural products 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 150000002191 fatty alcohols Chemical class 0.000 description 2
- 239000012737 fresh medium Substances 0.000 description 2
- 210000004907 gland Anatomy 0.000 description 2
- 230000012010 growth Effects 0.000 description 2
- 244000005709 gut microbiome Species 0.000 description 2
- 210000003405 ileum Anatomy 0.000 description 2
- 210000000987 immune system Anatomy 0.000 description 2
- 229960003444 immunosuppressant agent Drugs 0.000 description 2
- 239000003018 immunosuppressive agent Substances 0.000 description 2
- 230000008595 infiltration Effects 0.000 description 2
- 238000001764 infiltration Methods 0.000 description 2
- 210000004969 inflammatory cell Anatomy 0.000 description 2
- 230000005764 inhibitory process Effects 0.000 description 2
- 208000014674 injury Diseases 0.000 description 2
- 229940072205 lactobacillus plantarum Drugs 0.000 description 2
- 244000005700 microbiome Species 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 210000004877 mucosa Anatomy 0.000 description 2
- 210000004400 mucous membrane Anatomy 0.000 description 2
- 210000005036 nerve Anatomy 0.000 description 2
- 230000002093 peripheral effect Effects 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 229920000136 polysorbate Polymers 0.000 description 2
- 229950008882 polysorbate Drugs 0.000 description 2
- 230000001105 regulatory effect Effects 0.000 description 2
- YGSDEFSMJLZEOE-UHFFFAOYSA-N salicylic acid Chemical compound OC(=O)C1=CC=CC=C1O YGSDEFSMJLZEOE-UHFFFAOYSA-N 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 235000019333 sodium laurylsulphate Nutrition 0.000 description 2
- 239000000758 substrate Substances 0.000 description 2
- 150000003871 sulfonates Chemical class 0.000 description 2
- 239000006228 supernatant Substances 0.000 description 2
- 208000024891 symptom Diseases 0.000 description 2
- 230000009885 systemic effect Effects 0.000 description 2
- 210000004876 tela submucosa Anatomy 0.000 description 2
- 239000000080 wetting agent Substances 0.000 description 2
- JNYAEWCLZODPBN-JGWLITMVSA-N (2r,3r,4s)-2-[(1r)-1,2-dihydroxyethyl]oxolane-3,4-diol Chemical class OC[C@@H](O)[C@H]1OC[C@H](O)[C@H]1O JNYAEWCLZODPBN-JGWLITMVSA-N 0.000 description 1
- MZOFCQQQCNRIBI-VMXHOPILSA-N (3s)-4-[[(2s)-1-[[(2s)-1-[[(1s)-1-carboxy-2-hydroxyethyl]amino]-4-methyl-1-oxopentan-2-yl]amino]-5-(diaminomethylideneamino)-1-oxopentan-2-yl]amino]-3-[[2-[[(2s)-2,6-diaminohexanoyl]amino]acetyl]amino]-4-oxobutanoic acid Chemical compound OC[C@@H](C(O)=O)NC(=O)[C@H](CC(C)C)NC(=O)[C@H](CCCN=C(N)N)NC(=O)[C@H](CC(O)=O)NC(=O)CNC(=O)[C@@H](N)CCCCN MZOFCQQQCNRIBI-VMXHOPILSA-N 0.000 description 1
- ZORQXIQZAOLNGE-UHFFFAOYSA-N 1,1-difluorocyclohexane Chemical compound FC1(F)CCCCC1 ZORQXIQZAOLNGE-UHFFFAOYSA-N 0.000 description 1
- CMCBDXRRFKYBDG-UHFFFAOYSA-N 1-dodecoxydodecane Chemical compound CCCCCCCCCCCCOCCCCCCCCCCCC CMCBDXRRFKYBDG-UHFFFAOYSA-N 0.000 description 1
- IIZPXYDJLKNOIY-JXPKJXOSSA-N 1-palmitoyl-2-arachidonoyl-sn-glycero-3-phosphocholine Chemical compound CCCCCCCCCCCCCCCC(=O)OC[C@H](COP([O-])(=O)OCC[N+](C)(C)C)OC(=O)CCC\C=C/C\C=C/C\C=C/C\C=C/CCCCC IIZPXYDJLKNOIY-JXPKJXOSSA-N 0.000 description 1
- 229940114072 12-hydroxystearic acid Drugs 0.000 description 1
- TVZRAEYQIKYCPH-UHFFFAOYSA-N 3-(trimethylsilyl)propane-1-sulfonic acid Chemical compound C[Si](C)(C)CCCS(O)(=O)=O TVZRAEYQIKYCPH-UHFFFAOYSA-N 0.000 description 1
- WUBBRNOQWQTFEX-UHFFFAOYSA-N 4-aminosalicylic acid Chemical compound NC1=CC=C(C(O)=O)C(O)=C1 WUBBRNOQWQTFEX-UHFFFAOYSA-N 0.000 description 1
- XZIIFPSPUDAGJM-UHFFFAOYSA-N 6-chloro-2-n,2-n-diethylpyrimidine-2,4-diamine Chemical compound CCN(CC)C1=NC(N)=CC(Cl)=N1 XZIIFPSPUDAGJM-UHFFFAOYSA-N 0.000 description 1
- 241000186012 Bifidobacterium breve Species 0.000 description 1
- 241001608472 Bifidobacterium longum Species 0.000 description 1
- 241000193171 Clostridium butyricum Species 0.000 description 1
- 229920000858 Cyclodextrin Polymers 0.000 description 1
- 102000004127 Cytokines Human genes 0.000 description 1
- 108090000695 Cytokines Proteins 0.000 description 1
- 206010012735 Diarrhoea Diseases 0.000 description 1
- 208000036649 Dysbacteriosis Diseases 0.000 description 1
- 208000027244 Dysbiosis Diseases 0.000 description 1
- 241000194033 Enterococcus Species 0.000 description 1
- 241000520130 Enterococcus durans Species 0.000 description 1
- 241000194032 Enterococcus faecalis Species 0.000 description 1
- IAYPIBMASNFSPL-UHFFFAOYSA-N Ethylene oxide Chemical compound C1CO1 IAYPIBMASNFSPL-UHFFFAOYSA-N 0.000 description 1
- PIICEJLVQHRZGT-UHFFFAOYSA-N Ethylenediamine Chemical class NCCN PIICEJLVQHRZGT-UHFFFAOYSA-N 0.000 description 1
- 244000199866 Lactobacillus casei Species 0.000 description 1
- 235000013958 Lactobacillus casei Nutrition 0.000 description 1
- 206010024642 Listless Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 206010061297 Mucosal erosion Diseases 0.000 description 1
- 206010061298 Mucosal haemorrhage Diseases 0.000 description 1
- 206010028140 Mucous stools Diseases 0.000 description 1
- 108010019160 Pancreatin Proteins 0.000 description 1
- 208000007913 Pituitary Neoplasms Diseases 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 229920001214 Polysorbate 60 Polymers 0.000 description 1
- 241000588769 Proteus <enterobacteria> Species 0.000 description 1
- 241000246044 Sophora flavescens Species 0.000 description 1
- 241000194017 Streptococcus Species 0.000 description 1
- 241000194020 Streptococcus thermophilus Species 0.000 description 1
- 210000000447 Th1 cell Anatomy 0.000 description 1
- 102000002689 Toll-like receptor Human genes 0.000 description 1
- 108020000411 Toll-like receptor Proteins 0.000 description 1
- 230000006978 adaptation Effects 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- 150000003973 alkyl amines Chemical class 0.000 description 1
- 150000008051 alkyl sulfates Chemical class 0.000 description 1
- 230000004075 alteration Effects 0.000 description 1
- 150000001408 amides Chemical class 0.000 description 1
- 125000000539 amino acid group Chemical group 0.000 description 1
- 150000001413 amino acids Chemical group 0.000 description 1
- 229960004909 aminosalicylic acid Drugs 0.000 description 1
- 238000004458 analytical method Methods 0.000 description 1
- 125000000129 anionic group Chemical group 0.000 description 1
- 230000008485 antagonism Effects 0.000 description 1
- 230000006399 behavior Effects 0.000 description 1
- 230000003542 behavioural effect Effects 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- 230000008901 benefit Effects 0.000 description 1
- 229960000686 benzalkonium chloride Drugs 0.000 description 1
- CADWTSSKOVRVJC-UHFFFAOYSA-N benzyl(dimethyl)azanium;chloride Chemical compound [Cl-].C[NH+](C)CC1=CC=CC=C1 CADWTSSKOVRVJC-UHFFFAOYSA-N 0.000 description 1
- 229940009291 bifidobacterium longum Drugs 0.000 description 1
- 239000003833 bile salt Substances 0.000 description 1
- 229940093761 bile salts Drugs 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 210000004369 blood Anatomy 0.000 description 1
- 239000008280 blood Substances 0.000 description 1
- 208000027503 bloody stool Diseases 0.000 description 1
- 210000004556 brain Anatomy 0.000 description 1
- 230000005978 brain dysfunction Effects 0.000 description 1
- 230000003925 brain function Effects 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 150000003857 carboxamides Chemical class 0.000 description 1
- 150000007942 carboxylates Chemical class 0.000 description 1
- 150000001733 carboxylic acid esters Chemical class 0.000 description 1
- YIOHILISQLOJJV-UHFFFAOYSA-N carboxymethyl-dimethyl-octadec-9-enylazanium;hydroxide Chemical compound [OH-].CCCCCCCCC=CCCCCCCCC[N+](C)(C)CC(O)=O YIOHILISQLOJJV-UHFFFAOYSA-N 0.000 description 1
- 239000004359 castor oil Substances 0.000 description 1
- 235000019438 castor oil Nutrition 0.000 description 1
- 125000002091 cationic group Chemical group 0.000 description 1
- 230000024245 cell differentiation Effects 0.000 description 1
- 239000013592 cell lysate Substances 0.000 description 1
- 238000005119 centrifugation Methods 0.000 description 1
- 230000008859 change Effects 0.000 description 1
- 238000012512 characterization method Methods 0.000 description 1
- 239000002738 chelating agent Substances 0.000 description 1
- 239000003153 chemical reaction reagent Substances 0.000 description 1
- 239000003795 chemical substances by application Substances 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 206010009887 colitis Diseases 0.000 description 1
- 230000000112 colonic effect Effects 0.000 description 1
- 229940097362 cyclodextrins Drugs 0.000 description 1
- 235000013365 dairy product Nutrition 0.000 description 1
- DTPCFIHYWYONMD-UHFFFAOYSA-N decaethylene glycol Polymers OCCOCCOCCOCCOCCOCCOCCOCCOCCOCCO DTPCFIHYWYONMD-UHFFFAOYSA-N 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 229960003964 deoxycholic acid Drugs 0.000 description 1
- 238000001514 detection method Methods 0.000 description 1
- 238000010586 diagram Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 210000004921 distal colon Anatomy 0.000 description 1
- 229960000878 docusate sodium Drugs 0.000 description 1
- 125000003438 dodecyl group Chemical group [H]C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])* 0.000 description 1
- 231100000673 dose–response relationship Toxicity 0.000 description 1
- 239000012154 double-distilled water Substances 0.000 description 1
- 239000003937 drug carrier Substances 0.000 description 1
- 230000007140 dysbiosis Effects 0.000 description 1
- 229940032049 enterococcus faecalis Drugs 0.000 description 1
- 208000010227 enterocolitis Diseases 0.000 description 1
- 150000002148 esters Chemical class 0.000 description 1
- 238000011156 evaluation Methods 0.000 description 1
- 150000004665 fatty acids Chemical class 0.000 description 1
- 238000004108 freeze drying Methods 0.000 description 1
- 230000002496 gastric effect Effects 0.000 description 1
- 239000003629 gastrointestinal hormone Substances 0.000 description 1
- 239000003862 glucocorticoid Substances 0.000 description 1
- 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 1
- 150000002334 glycols Chemical class 0.000 description 1
- 210000002175 goblet cell Anatomy 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 208000035861 hematochezia Diseases 0.000 description 1
- 235000015092 herbal tea Nutrition 0.000 description 1
- 238000004128 high performance liquid chromatography Methods 0.000 description 1
- 239000005556 hormone Substances 0.000 description 1
- 229940088597 hormone Drugs 0.000 description 1
- 230000036737 immune function Effects 0.000 description 1
- 230000001861 immunosuppressant effect Effects 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 208000027866 inflammatory disease Diseases 0.000 description 1
- 230000002401 inhibitory effect Effects 0.000 description 1
- 239000002054 inoculum Substances 0.000 description 1
- 230000007358 intestinal barrier function Effects 0.000 description 1
- 230000008944 intestinal immunity Effects 0.000 description 1
- 230000003870 intestinal permeability Effects 0.000 description 1
- 230000002262 irrigation Effects 0.000 description 1
- 238000003973 irrigation Methods 0.000 description 1
- 229940017800 lactobacillus casei Drugs 0.000 description 1
- 235000010445 lecithin Nutrition 0.000 description 1
- 239000000787 lecithin Substances 0.000 description 1
- 229940067606 lecithin Drugs 0.000 description 1
- 231100000518 lethal Toxicity 0.000 description 1
- 230000001665 lethal effect Effects 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 208000017971 listlessness Diseases 0.000 description 1
- 239000002932 luster Substances 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 230000007246 mechanism Effects 0.000 description 1
- 210000004379 membrane Anatomy 0.000 description 1
- 239000012528 membrane Substances 0.000 description 1
- WSFSSNUMVMOOMR-NJFSPNSNSA-N methanone Chemical compound O=[14CH2] WSFSSNUMVMOOMR-NJFSPNSNSA-N 0.000 description 1
- 238000000034 method Methods 0.000 description 1
- 230000006724 microglial activation Effects 0.000 description 1
- 238000002156 mixing Methods 0.000 description 1
- 210000001616 monocyte Anatomy 0.000 description 1
- 238000010172 mouse model Methods 0.000 description 1
- PSZYNBSKGUBXEH-UHFFFAOYSA-N naphthalene-1-sulfonic acid Chemical class C1=CC=C2C(S(=O)(=O)O)=CC=CC2=C1 PSZYNBSKGUBXEH-UHFFFAOYSA-N 0.000 description 1
- 229920000847 nonoxynol Polymers 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 230000035764 nutrition Effects 0.000 description 1
- QIQXTHQIDYTFRH-UHFFFAOYSA-N octadecanoic acid Chemical compound CCCCCCCCCCCCCCCCCC(O)=O QIQXTHQIDYTFRH-UHFFFAOYSA-N 0.000 description 1
- 229940066429 octoxynol Drugs 0.000 description 1
- 229920002113 octoxynol Polymers 0.000 description 1
- 235000014593 oils and fats Nutrition 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- JRZJOMJEPLMPRA-UHFFFAOYSA-N olefin Natural products CCCCCCCC=C JRZJOMJEPLMPRA-UHFFFAOYSA-N 0.000 description 1
- 150000007524 organic acids Chemical class 0.000 description 1
- 230000003647 oxidation Effects 0.000 description 1
- 238000007254 oxidation reaction Methods 0.000 description 1
- 230000033116 oxidation-reduction process Effects 0.000 description 1
- 229910052760 oxygen Inorganic materials 0.000 description 1
- 239000001301 oxygen Substances 0.000 description 1
- 239000003002 pH adjusting agent Substances 0.000 description 1
- 125000000913 palmityl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 229940055695 pancreatin Drugs 0.000 description 1
- FJKROLUGYXJWQN-UHFFFAOYSA-N papa-hydroxy-benzoic acid Natural products OC(=O)C1=CC=C(O)C=C1 FJKROLUGYXJWQN-UHFFFAOYSA-N 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 230000001717 pathogenic effect Effects 0.000 description 1
- 231100000255 pathogenic effect Toxicity 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 210000005259 peripheral blood Anatomy 0.000 description 1
- 239000011886 peripheral blood Substances 0.000 description 1
- 230000008855 peristalsis Effects 0.000 description 1
- 239000012466 permeate Substances 0.000 description 1
- 238000005502 peroxidation Methods 0.000 description 1
- 239000003208 petroleum Substances 0.000 description 1
- 239000000546 pharmaceutical excipient Substances 0.000 description 1
- 239000008180 pharmaceutical surfactant Substances 0.000 description 1
- 239000008389 polyethoxylated castor oil Substances 0.000 description 1
- 229920001223 polyethylene glycol Polymers 0.000 description 1
- 229920000642 polymer Polymers 0.000 description 1
- 229920000056 polyoxyethylene ether Polymers 0.000 description 1
- 230000034190 positive regulation of NF-kappaB transcription factor activity Effects 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 239000003755 preservative agent Substances 0.000 description 1
- 230000000770 proinflammatory effect Effects 0.000 description 1
- 230000001737 promoting effect Effects 0.000 description 1
- 102000004169 proteins and genes Human genes 0.000 description 1
- 108090000623 proteins and genes Proteins 0.000 description 1
- 150000003242 quaternary ammonium salts Chemical class 0.000 description 1
- 230000007115 recruitment Effects 0.000 description 1
- 210000000664 rectum Anatomy 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000008929 regeneration Effects 0.000 description 1
- 238000011069 regeneration method Methods 0.000 description 1
- 238000004007 reversed phase HPLC Methods 0.000 description 1
- 229960004889 salicylic acid Drugs 0.000 description 1
- 210000000813 small intestine Anatomy 0.000 description 1
- 229910052708 sodium Inorganic materials 0.000 description 1
- 239000011734 sodium Substances 0.000 description 1
- NRHMKIHPTBHXPF-TUJRSCDTSA-M sodium cholate Chemical compound [Na+].C([C@H]1C[C@H]2O)[C@H](O)CC[C@]1(C)[C@@H]1[C@@H]2[C@@H]2CC[C@H]([C@@H](CCC([O-])=O)C)[C@@]2(C)[C@@H](O)C1 NRHMKIHPTBHXPF-TUJRSCDTSA-M 0.000 description 1
- FHHPUSMSKHSNKW-SMOYURAASA-M sodium deoxycholate Chemical compound [Na+].C([C@H]1CC2)[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 FHHPUSMSKHSNKW-SMOYURAASA-M 0.000 description 1
- APSBXTVYXVQYAB-UHFFFAOYSA-M sodium docusate Chemical compound [Na+].CCCCC(CC)COC(=O)CC(S([O-])(=O)=O)C(=O)OCC(CC)CCCC APSBXTVYXVQYAB-UHFFFAOYSA-M 0.000 description 1
- 159000000000 sodium salts Chemical class 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000000243 solution Substances 0.000 description 1
- 229940035044 sorbitan monolaurate Drugs 0.000 description 1
- 235000011069 sorbitan monooleate Nutrition 0.000 description 1
- 239000001593 sorbitan monooleate Substances 0.000 description 1
- 229940035049 sorbitan monooleate Drugs 0.000 description 1
- 229950011392 sorbitan stearate Drugs 0.000 description 1
- 125000004079 stearyl group Chemical group [H]C([*])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])C([H])([H])[H] 0.000 description 1
- 150000003431 steroids Chemical class 0.000 description 1
- 210000002784 stomach Anatomy 0.000 description 1
- 239000000126 substance Substances 0.000 description 1
- 238000006467 substitution reaction Methods 0.000 description 1
- 230000004083 survival effect Effects 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 229940037128 systemic glucocorticoids Drugs 0.000 description 1
- 238000004885 tandem mass spectrometry Methods 0.000 description 1
- 125000003831 tetrazolyl group Chemical group 0.000 description 1
- 238000010257 thawing Methods 0.000 description 1
- 229920001664 tyloxapol Polymers 0.000 description 1
- MDYZKJNTKZIUSK-UHFFFAOYSA-N tyloxapol Chemical compound O=C.C1CO1.CC(C)(C)CC(C)(C)C1=CC=C(O)C=C1 MDYZKJNTKZIUSK-UHFFFAOYSA-N 0.000 description 1
- 229960004224 tyloxapol Drugs 0.000 description 1
- 238000000108 ultra-filtration Methods 0.000 description 1
- 238000010200 validation analysis Methods 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 1
- 230000004580 weight loss Effects 0.000 description 1
- 238000001262 western blot Methods 0.000 description 1
Classifications
-
- C—CHEMISTRY; METALLURGY
- C07—ORGANIC CHEMISTRY
- C07K—PEPTIDES
- C07K14/00—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- C07K14/415—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- 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
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
- A61K38/16—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof
- A61K38/168—Peptides having more than 20 amino acids; Gastrins; Somatostatins; Melanotropins; Derivatives thereof from plants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P1/00—Drugs for disorders of the alimentary tract or the digestive system
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P29/00—Non-central analgesic, antipyretic or antiinflammatory agents, e.g. antirheumatic agents; Non-steroidal antiinflammatory drugs [NSAID]
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P37/00—Drugs for immunological or allergic disorders
- A61P37/02—Immunomodulators
- A61P37/04—Immunostimulants
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K38/00—Medicinal preparations containing peptides
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Medicinal Chemistry (AREA)
- General Health & Medical Sciences (AREA)
- Public Health (AREA)
- Organic Chemistry (AREA)
- Pharmacology & Pharmacy (AREA)
- Animal Behavior & Ethology (AREA)
- Veterinary Medicine (AREA)
- Immunology (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Engineering & Computer Science (AREA)
- Microbiology (AREA)
- Gastroenterology & Hepatology (AREA)
- Proteomics, Peptides & Aminoacids (AREA)
- Mycology (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Botany (AREA)
- Rheumatology (AREA)
- Pain & Pain Management (AREA)
- Biochemistry (AREA)
- Biophysics (AREA)
- Genetics & Genomics (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Medicines That Contain Protein Lipid Enzymes And Other Medicines (AREA)
Abstract
本发明涉及一种提高免疫力的益生菌组合物以及在治疗疾病中的应用。所述益生菌组合物是由加氏乳杆菌TCI515、鼠李糖乳杆菌菌株和瑞士乳杆菌和抗炎肽SF‑R‑4组成。本发明的益生菌组合物能够有效的降低溃疡性结肠炎小鼠的DAI积分和组织学评分，具有较好的治疗效果。
Description
技术领域
本申请涉及生物领域，具体的涉及一种提高免疫力的益生菌组合物以及在治疗疾病中的应用。
背景技术
炎性肠病(inflammatory bowel disease，IBD)是一组病因尚未阐明的慢性非特异性肠道炎性疾病，包括溃疡性结肠炎(UC)和克罗恩病(CD)。UC主要为局限于黏膜和黏膜下层的连续性炎性反应，病变多自直肠开始，可累及全结肠甚至末段回肠；CD则为节段性的肉芽肿性炎性反应，可累及消化道任何部位，但多常见于末段回肠和邻近结肠。IBD的传统治疗方法包括氨基水杨酸制剂、糖皮质激素、免疫抑制剂、生物制剂等，都可抑制肠道炎性反应，但存在价格高、安全性低、不良反应明显、治疗作用有限等缺点。
益生菌种类繁多，人体内就有超过400种。从目前已报道的益生菌来看，大致可分为以下5类：乳杆菌类，如干酪乳杆菌、植物乳杆菌、嗜酸乳杆菌等；双歧杆菌类，如长双歧杆菌、短双歧杆菌等；芽孢杆菌类，如布拉地芽孢杆菌、丁酸梭状芽孢杆菌等；链球菌属:如嗜热链球菌；肠球菌属，如耐久肠球菌、粪肠球菌等。已有大量研究证实，益生菌在缓解或治疗IBD中起到重要作用。有研究表明，益生菌可提升肠道黏膜自身预防机制，且可拮抗致病菌的致病作用，还可以控制炎性因子的释放及短时间内恢复肠道菌群。肠道菌群失调是促进IBD发生与发展的一个重要因素，研究发现，益生菌能够通过Toll样受体促进Th1细胞分化，进而改变黏膜免疫系统，增强肠道保护功能，增加肠道菌群多样性。有研究表明，植物乳杆菌Q7的细胞外囊泡(EV)可调节肠道微生物群，增加肠道中乳酸杆菌、双歧杆菌等有益细菌的丰度并降低变形杆菌等有害细菌的丰度。益生菌还可通过竞争营养、拮抗等方式与肠道微生物群相互作用，降低致病菌活性，以及产生某些抑菌性物质，如有机酸，直接抑制病原菌的生长。上述研究表明，益生菌可以通过调节肠道菌群影响IBD的发生和发展。
类似IBD的全身性炎性疾病患者往往伴有全身虚弱等症状，这可能是由于大脑功能的改变，而益生菌可以通过神经、激素和免疫系统的变化改善“微生物-脑-肠”轴的功能，进而改善上述症状。外周TNF-a可以诱导小胶质细胞激活和募集单核细胞到大脑，这是影响炎症相关的疾病行为发展的关键环节，而使用混合益生菌可以降低动物模型外周血的TNF-a水平，提示益生菌可调节外周炎症(结肠炎)相关的大脑功能障碍和行为改变。越来越多的研究证实益生菌在调节肠道菌群、调节肠道免疫，减轻肠道炎性反应、增加机体抗氧化能力、降低肠道通透性、增强肠道屏障功能等方面发挥重要作用，益生菌或可成为缓解或治疗IBD的新手段。但目前仍有很多问题需要解决，如益生菌的给药剂量、给药间隔及疗程等，且其不良反应尚存在争议，未来需要更多的临床试验进行进一步探究。
此外，目前临床上治疗炎症性肠病还有其他常用的三类药：水杨酸类、类固醇类和免疫抑制剂，对溃疡性结肠炎和克罗恩病的缓解率分别为80％和70％左右，且停药后易复发。研究发现，溃疡性结肠炎患者中GAP-43没有变化，因为GAP-43是与神经再生相关的，可以反映神经元再生的发生，由此推断溃疡性结肠炎患者SP的升高与肠神经元的再生没有明显关系，亦即溃疡性结肠炎中没有明显的肠神经元再生，而SP的升高可能与SP的合成增多有关。进一步的，多肽类药物也有用于炎症性肠病的治疗，比如铃蟾肽是一种含有14个氨基酸残基的生物活性肽。铃蟾肽能刺激其他几种胃肠激素的释放,调节胃肠蠕动,刺激消化道正常粘膜组织的生长,提高致死性小肠结肠炎动物的存活率。但是该多肽并非具有特异性的治疗方向，效果也有待进一步的提高。
总之，目前还没有疗效较好，性价比更高的较好的用于治疗益生菌组合物的出现。
发明内容
本发明克服现有技术的缺陷，提供了一种特异性拥有治疗炎症性肠病同时提高免疫力的益生菌组合物。
所述的益生菌包括鼠李糖乳杆菌菌株和瑞士乳杆菌 前者是一种乳杆菌，它于1976年首次从乳制品中分离出来，鼠李糖乳杆菌Rosell11经常与瑞士乳杆菌R52一起研究，通常被称为“Lacidofil”组合。这两种菌株都列在EFSA的合格安全菌株(QPS)列表中。所述菌株具有提高免疫功能的功能，对于改善肠道特性也有一定功效。二个菌株都是现有技术。
进一步的，所述的益生菌还包含来自大江生医股份有限公司的加氏乳杆菌TCI515，该菌株是保藏在DSMZ德国微生物保藏中心(German Collection ofMicroorganisms and Cell Cultures，DSMZ)的DSM33105菌株。
所述的益生菌组合物是由提高免疫力的益生菌和治疗炎症性肠病的多肽。
进一步的，所述的多肽是分离自苦参。
苦参是一种中药材，具有抗菌和抗炎特性。被广泛应用于中医治疗肠炎等疾病。苦参的提取物被用于制成草药茶或药膏，可以减轻肠炎引起的炎症和腹泻。发明人将苦参粉碎后通过胰酶水解后用膜过滤去掉固体组分，通过超滤截留高活性分子量小玉3Kda的渗透物，采用RP-HPLC分离抗炎活性多肽，经过分离的HPLC组分再经LS-MS/MS鉴定获得了活性最高的抗炎肽SF-R-4，其氨基酸序列如SEQ ID NO：1所示。
本发明提供了一种提高免疫力治疗炎症性肠病的益生菌组合物，所述益生菌组合物是按照加氏乳杆菌TCI515、鼠李糖乳杆菌菌株和瑞士乳杆菌的比例分别为2:1:1的重量比例来制备获得益生菌组分，同时按照益生菌和抗炎肽SF-R-4质量比为95:5的比例添加多肽，最终制备获得益生菌组合物。
进一步的，本发明还提供了抗炎肽SF-R-4在制备用于治疗炎症性肠病的药物中用途。
进一步的，本发明还提供了含有抗炎肽SF-R-4的益生菌组合物在制备用于提高免疫力治疗炎症性肠病的药剂中的用途。所述的所述益生菌组合物是按照加氏乳杆菌TCI515、鼠李糖乳杆菌菌株和瑞士乳杆菌的比例分别为2:1:1的重量比例来制备获得益生菌组分，同时按照益生菌和抗炎肽SF-R-4质量比为95:5的比例添加多肽，最终制备获得益生菌组合物。
进一步的，本发明的药物或者组合物中还含有其他药学上可接受的载体。
适合的药学上可接受的赋形剂的实例包括一种或多种聚合物、润湿剂或表面活性剂、pH调节剂、等渗调节剂、防腐剂、缓冲剂以及螯合剂、或它们的任意组合。
适合的药学上可接受的润湿剂或表面活性剂的实例包括但不限于两性、非离子型、阳离子型或阴离子型分子。适合的表面活性剂包括但不限于聚山梨酯、十二烷基硫酸钠(月桂基硫酸钠)、十二烷基二甲基氧化胺、多库酯钠、十六烷基三甲基溴化铵(CTAB)、聚乙氧基化醇、聚氧乙烯脱水山梨醇、辛苯昔醇、N,N-二甲基十二烷基胺-N-氧化物、十六烷基三甲基溴化铵、聚氧乙烯(10)月桂醚、表面活性剂(由月桂醇、鲸蜡醇、硬脂醇和油醇衍生的植物油系脂肪醇聚氧乙烯醚)、胆盐(例如脱氧胆酸钠和胆酸钠)、聚氧乙烯蓖麻油、壬基酚乙氧基化物、环糊精、卵磷脂、甲苄索氯铵、羧酸盐、磺酸盐、石油磺酸盐、烷基苯磺酸盐、萘磺酸盐、烯烃磺酸盐、烷基硫酸盐、硫酸盐、硫酸化天然油和脂肪、硫酸化酯、硫酸化烷醇酰胺、烷基酚(乙氧基化和硫酸化的)、乙氧基化脂肪醇、聚氧乙烯表面活性剂、羧酸酯、聚乙二醇酯、脱水山梨醇酯及其乙氧基化衍生物、脂肪酸乙二醇酯、羧酰胺、单烷醇胺缩合物、聚氧乙烯脂肪酸酰胺、季铵盐、具有酰胺键的胺、聚氧乙烯烷基胺和聚氧乙烯脂环族胺、N,N,N,N-四取代乙二胺、2-烷基-1-羟乙基-2-咪唑啉、N-椰油基-3-氨基丙酸/钠盐、N-牛脂基-3-亚氨基二丙酸二钠盐、N-羧甲基-N,N-二甲基-N-9-十八烯基氢氧化铵、N-椰油酰胺乙基-N-羟乙基甘氨酸钠盐等、聚氧乙烯、脱水山梨醇单月桂酸酯和硬脂酸酯、(聚乙氧基化蓖麻油)、(环氧乙烷/12-羟基硬脂酸)、聚山梨酯、四丁酚醛，以及它们的任意组合。优选的药学上可接受的表面活性剂包括四丁酚醛和(脱水山梨醇单油酸酯)或其混合物。
有益效果
本发明提供了含有抗炎肽SF-R-4的益生菌组合物在制备用于提高免疫力治疗炎症性肠病的药剂中的用途。所述的所述益生菌组合物是由加氏乳杆菌TCI515、鼠李糖乳杆菌菌株和瑞士乳杆菌和抗炎肽SF-R-4组成。本发明的益生菌组合物能够有效的降低溃疡性结肠炎小鼠的DAI积分和组织学评分，具有较好的治疗效果。
附图说明
图1多肽抗炎效果鉴定结果图
图2多肽对NF-κB以及MAPK信号通路激活的影响结果图
具体实施方式
下面将参照附图更详细地描述本发明的具体实施例。虽然附图中显示了本发明的具体实施例，然而应当理解，可以以各种形式实现本发明而不应被这里阐述的实施例所限制。相反，提供这些实施例是为了能够更透彻地理解本发明，并且能够将本发明的范围完整的传达给本领域的技术人员。
实施例1抗炎肽功能鉴定
人小肠粘膜上皮细胞使用人小肠粘膜上皮细胞完全培养基PDHX-G138，置于恒温细胞培养箱培养，培养箱条件是37℃，5％CO2，95％湿度条件。用于构建细胞炎症模型的细胞的细胞代数为18-20代。将细胞培养4-5天，直至形成融合，每2天半量更换新鲜培养基。
通过向融合状态下的细胞培养物中加入2ng/mL TNF-α诱导炎症。将小肠粘膜上皮细胞以1×105/mL的密度接种在48孔培养板中，并培养4-5天，每2天半量更换新鲜培养基。用相同的培养基配制的不同浓度的抗炎肽SF-R-4(终浓度分别为0.1mg/mL、0.5mg/mL和1mg/mL)孵育2小时。用磷酸盐缓冲液(PBS)洗涤细胞，加入终浓度为2ng/mL TNF-α孵育5小时。6小时后，收集上清液用于后续指标测定。每个平板含有阴性对照(仅用培养基处理)和阳性对照(用培养基和TNF-α处理)。
用培养基以1:30比例稀释水溶性四唑盐(WST-1)试剂。样品和细胞孵育完后，去除细胞上清液，用PBS洗涤3次，每孔加入200μL稀释后的WST-1，并在37℃下孵育10分钟，监测颜色变化，使用酶标仪测定其在450nm处的OD来检测IL-8的相对表达量，以模型组的IL-8作为1。结果如图1所示。
从图1可以看出，单独加入2ng/mL TNF-α孵育5小时后IL-8分泌显著增加(p<0.05)，表明体外炎症模型成功建立。用抗炎肽SF-R-4处理以剂量依赖的方式显著降低TNF-α诱导的人小肠粘膜上皮细胞IL-8分泌，在浓度为1mg/mL时抑制IL-8的效率达到了(38.13±1.89)％，效果显著。说明该多肽具有较好的抗炎活性。
进一步的，使用磷酸化-JNK，JNK，磷酸化-p38，p38，磷酸化-IκB和IκB的抗体孵育，进行各组(阴性组，模型组和1mg/mL多肽组)细胞裂解物的蛋白质印迹分析。使用ImageJ软件定量分析磷酸化的JNK，IκB和p38的相对水平，结果表示为与未处理的细胞相比的相对蛋白质水平，分析了对TNF-α诱导的细胞中NF-κB以及MAPK信号通路激活的影响。结果如图2所示。
如图2所示，与阴性组相比模型组的p38、IκB和JNK的磷酸化显著增加。结果表明TNF-α可以激活NF-κB以及MAPK信号通路，从而上调促炎细胞因子的表达。均能显著抑制JNK的磷酸化。然而，用抗炎肽SF-R-4预处理能还显著抑制TNF-α诱导的IκB以及p38磷酸化，能够轻微抑制JNK的磷酸化。结果表明，抗炎肽SF-R-4对IκB、p38磷酸化的抑制可以有效阻断NF-κB和MAPK信号通路的激活从而抗炎和提高免疫力。
实施例2益生菌组合物的制备
分别将加氏乳杆菌TCI515、鼠李糖乳杆菌菌株和瑞士乳杆菌解冻及活化后，依1％的接种量在37℃静置培养于乳杆菌属MRSD培养基(55g乳杆菌MRS肉汤，溶于1L去离子水，pH值约6.8)18小时。所得细菌培养物经过5000rpm离心20分钟后分别获得三种单独的菌体。将三种菌体分别冷冻干燥后得到冻干菌粉，按照加氏乳杆菌TCI515、鼠李糖乳杆菌 菌株和瑞士乳杆菌的比例分别为2:1:1的重量比例来制备获得益生菌组分，同时按照益生菌和抗炎肽SF-R-4质量比为95:5的比例添加多肽，最终制备获得益生菌组合物。
实施例3益生菌组合物以及多肽的功效验证
取雄性Balb/c鼠随机分组，分别为正常组、模型组、阴性对照(生理盐水)组、阳性对照美沙拉嗪组、本发明含多肽益生菌组、本发明不含多肽益生菌组以及SF-R-4多肽组，每组10只。正常组自由饮用双重蒸馏水，其余组小鼠自由饮用2.5％DSS 7d建立急性溃疡性结肠炎动物模型。从造模前2d开始，阴性组、美沙拉嗪组、嗜酸乳杆菌低、高剂量组、嗜酸乳杆菌联合美沙拉嗪组分别给予无菌生理盐水、美沙拉嗪(30mg/ml)、含多肽益生菌组(30mg/ml)、不含多肽益生菌组(30mg/ml)不含多肽益生菌组(30mg/ml)、SF-R-4多肽组(1.5mg/ml)灌胃，每次0.2ml，早晚各1次。造模第8d上午折颈法处死小鼠，取出结肠，取部分远端结肠组织甲醛固定、石蜡包埋、切片，HE染色。疾病活动指数(DAI)积分：按照CotrolledPathological study and preoperative MRI evaluation of the consistency ofpituitary adenomas标准进行疾病活动指数(DAI)积分评定，结果如表1所示。
表1各组对小鼠的DAI积分
| 组别 | DAI评分 |
| 正常组 | 0 |
| 模型组 | 6.84±0.76 |
| 阴性对照组 | 6.82±0.82 |
| 阳性对照美沙拉嗪组 | 4.37±0.59* |
| 含多肽益生菌组 | 1.05±0.34** |
| 不含多肽益生菌组 | 5.12±0.68* |
| SF-R-4多肽组 | 2.17±0.41** |
*表示与模型组比较，差异显著P<0.05,**表示与模型组比较，差异极其显著P<0.01。
结果显示正常对照组小鼠在饮食、活动及大便性状方面均正常，模型小鼠在造模后出现体重减轻、毛发无光泽、活动减少、精神萎靡、粘液便，严重者可见肉眼血便。正常组小鼠DAI积分始终维持在零水平，与各组比较差异有统计学意义；采用本发明的多肽或者采用多肽和益生菌组合，能够有效的降低DAI积分实现较好的治疗效果。
组织学损伤评分：按照Dielemen等评分标准。炎症：无(0分)，轻度(1分)，重度(2分)；病变深度：无(0分)，粘膜下层(1分)，肌层(2分)，浆膜层(3分)；隐窝破坏：无(0分)，基底1/3隐窝被破坏(1分)，基底2/3隐窝被破坏(2分)，仅有完整的表面上皮(3分)，全部隐窝和上皮被破坏(4分)；病变范围(％)：1-25计1分，26-50计2分，51-75计3分，76-100计4分。每个切片随机选取10个以上高倍视野(400倍)进行计分，取平均值作为组织学损伤计分。结果如表2所示。
表2各组对小鼠的组织学评分
| 组别 | DAI评分 |
| 正常组 | 0 |
| 模型组 | 10.57±1.53 |
| 阴性对照组 | 8.63±1.21 |
| 阳性对照美沙拉嗪组 | 5.10±0.84* |
| 含多肽益生菌组 | 1.87±0.46** |
| 不含多肽益生菌组 | 6.08±0.94* |
| SF-R-4多肽组 | 2.96±0.52** |
*表示与模型组比较，差异显著P<0.05,**表示与模型组比较，差异极其显著P<0.01。
通过分析发现，正常组小鼠结肠粘膜结构完整，腺体排列整齐，隐窝正常，杯状细胞无减少，未见粘膜糜烂、出血，无炎性细胞浸润。模型对照组与阴性对照组，结肠粘膜缺失，腺体不完整，有炎性细胞浸润，呈典型炎症改变，评分与正常组相比差异及其显著(P<0.01)。阳性对照组、含多肽益生菌组、不含多肽益生菌组和SF-R-4多肽组组织学评分降低，与模型和阴性对照组比较差异有统计学意义(P＜0.05)，见表2。从表2可以看出，本发明的多肽或者含有多肽的益生菌组能够极其显著的降低组织学评分，具有较好的治疗效果。
此外，通过检测小鼠小鼠血液中的MDA指标作为反应组织氧化还原状态和氧化应激情况的指标，可以发现，含多肽益生菌组和不含多肽益生菌组相比于模型组，MDA含量下降分别为53.5％和43.6％，具有较好的减轻组织内过氧化压力，氧自由基生成造成的损伤。说明多肽和益生菌一起能够较好的提高抗氧化免疫力的效果。
以上所述仅是本发明的优选实施方式，应当指出，对于本技术领域的普通技术人员来说，在不脱离本发明原理的前提下，还可以做出若干改进和润饰，这些改进和润饰也应视为本发明的保护范围。最后应说明的是：以上各实施例仅用以说明本发明的技术方案，而非对其限制；尽管参照前述各实施例对本发明进行了详细的说明，但本领域的普通技术人员应当理解：其依然可以对前述各实施例所记载的技术方案进行修改，或者对其中部分或者全部技术特征进行等同替换；而这些修改或者替换，并不使相应技术方案的本质脱离本发明各实施例技术方案的范围。
Claims (4)
1.一种抗炎肽SF-R-4，其特征在于：所述抗炎肽SF-R-4的氨基酸序列如SEQ ID NO：1所示。
2.抗炎肽SF-R-4在制备用于治疗炎症性肠病的药物中的用途，其中所述抗炎肽SF-R-4的氨基酸序列如SEQ ID NO：1所示。
3.一种用于提高免疫力治疗炎症性肠病的益生菌组合物，其特征在于所述益生菌组合物是按照加氏乳杆菌TCI515、鼠李糖乳杆菌Rosell®-11菌株和瑞士乳杆菌Roseell®-52的比例分别为2:1:1的重量比例来制备获得益生菌组分，同时按照益生菌组分和抗炎肽SF-R-4质量比为95:5的比例添加多肽，最终制备获得益生菌组合物；所述的抗炎肽SF-R-4的氨基酸序列如SEQ ID NO：1所示。
4.益生菌和多肽在制备用于提高免疫力治疗炎症性肠病的益生菌组合物中的用途，其中所述益生菌是按照加氏乳杆菌TCI515、鼠李糖乳杆菌Rosell®-11菌株和瑞士乳杆菌Roseell®-52的比例分别为2:1:1的重量比例来制备获得益生菌组分，同时按照益生菌组分和抗炎肽SF-R-4质量比为95:5的比例添加多肽，最终制备获得益生菌组合物；所述的抗炎肽SF-R-4的氨基酸序列如SEQ ID NO：1所示。
Priority Applications (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410149675.6A CN117986337B (zh) | 2024-02-02 | 2024-02-02 | 一种提高免疫力的益生菌组合物以及在治疗疾病中的应用 |
Applications Claiming Priority (1)
| Application Number | Priority Date | Filing Date | Title |
|---|---|---|---|
| CN202410149675.6A CN117986337B (zh) | 2024-02-02 | 2024-02-02 | 一种提高免疫力的益生菌组合物以及在治疗疾病中的应用 |
Publications (2)
| Publication Number | Publication Date |
|---|---|
| CN117986337A CN117986337A (zh) | 2024-05-07 |
| CN117986337B true CN117986337B (zh) | 2024-08-16 |
Family
ID=90890192
Family Applications (1)
| Application Number | Title | Priority Date | Filing Date |
|---|---|---|---|
| CN202410149675.6A Active CN117986337B (zh) | 2024-02-02 | 2024-02-02 | 一种提高免疫力的益生菌组合物以及在治疗疾病中的应用 |
Country Status (1)
| Country | Link |
|---|---|
| CN (1) | CN117986337B (zh) |
Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102665749A (zh) * | 2009-07-13 | 2012-09-12 | 不二制油株式会社 | 口服抗炎剂和口服抗炎肽 |
| CN116179427A (zh) * | 2022-12-27 | 2023-05-30 | 深圳科兴药业有限公司 | 一种益生菌组合物及其在制备具有抗结肠炎作用的产品中的应用 |
Family Cites Families (11)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CA2343840C (en) * | 1998-10-20 | 2011-03-08 | Vlaams Interuniversitair Instituut Voor Biotechnologie Vzw | Use of a cytokine-producing lactococcus strain to treat colitis |
| FI113741B (fi) * | 1999-11-01 | 2004-06-15 | Valio Oy | Menetelmä verenpainetta alentavia peptidejä sisältävän tuotteen valmistamiseksi |
| US20130305398A1 (en) * | 2012-02-16 | 2013-11-14 | Marie Coffin | Genes and uses for plant enhacement |
| BG66608B1 (bg) * | 2011-05-04 | 2017-10-16 | "Ел Би Булгарикум" ЕАД | Полибактериален пробиотичен препарат |
| EP3712173B1 (en) * | 2011-12-05 | 2023-07-12 | X-Body, Inc. | Pdgf receptor beta binding polypeptides |
| KR20150014483A (ko) * | 2012-05-11 | 2015-02-06 | 주식회사 카엘젬백스 | 항염증 활성을 갖는 펩티드 및 이를 포함하는 조성물 |
| EP3322430B1 (en) * | 2015-07-16 | 2020-03-25 | Nuritas Limited | Anti-inflammatory peptides, and uses thereof |
| US20180333442A1 (en) * | 2017-05-18 | 2018-11-22 | The Regents Of The University Of Michigan | Probiotic compositions and methods of use thereof |
| CN113493491B (zh) * | 2020-04-08 | 2023-07-04 | 清华大学 | 一种用于预防或治疗溃疡性结肠炎的多肽 |
| KR20210147362A (ko) * | 2020-05-28 | 2021-12-07 | 주식회사 미토퀘스트 | Nlrx1 유래의 신규한 항염증성 펩타이드 및 이의 용도 |
| CN115894612A (zh) * | 2022-09-06 | 2023-04-04 | 宁波大学 | 一种鸭肝蛋白源抗炎和抗菌功能肽及其制备方法和应用 |
-
2024
- 2024-02-02 CN CN202410149675.6A patent/CN117986337B/zh active Active
Patent Citations (2)
| Publication number | Priority date | Publication date | Assignee | Title |
|---|---|---|---|---|
| CN102665749A (zh) * | 2009-07-13 | 2012-09-12 | 不二制油株式会社 | 口服抗炎剂和口服抗炎肽 |
| CN116179427A (zh) * | 2022-12-27 | 2023-05-30 | 深圳科兴药业有限公司 | 一种益生菌组合物及其在制备具有抗结肠炎作用的产品中的应用 |
Also Published As
| Publication number | Publication date |
|---|---|
| CN117986337A (zh) | 2024-05-07 |
Similar Documents
| Publication | Publication Date | Title |
|---|---|---|
| CN109789173B (zh) | 源自芽孢杆菌细菌的纳米囊泡及其用途 | |
| US8092793B2 (en) | Treating inflammatory bowel disease with live bacteria | |
| CN114727640A (zh) | 细菌菌株、其组合物以及其用于治疗胃肠紊乱的用途 | |
| CN116555076B (zh) | 一株长双歧杆菌长亚种my1及其在制备通便和护肠食品药品中的应用 | |
| CN114259056A (zh) | 一种鼠李糖乳酪杆菌在制备用于预防和/或治疗溃疡性结肠炎的食品或药品中的应用 | |
| WO2017020784A1 (zh) | 一种脆弱拟杆菌及其应用 | |
| JP7126004B2 (ja) | 組成物及びその使用 | |
| CN117004503B (zh) | 一株唾液联合乳杆菌mb1及其在制备助睡眠和调理肠胃食品药品中的应用 | |
| CN115216423B (zh) | 一种动物双歧杆菌sf及其在医药和食品中的应用 | |
| CN118360221B (zh) | 一株卷曲乳杆菌ncu-23及其应用 | |
| CN118652782A (zh) | 一种凝结魏茨曼氏菌bcg44及其在体重控制、改善肠道健康和免疫功能中的应用 | |
| CN117987297B (zh) | 一株肠道菌及其在抗肿瘤免疫治疗中的应用 | |
| CN117384790B (zh) | 一株戊糖片球菌ks5及其在制备助睡眠药品中的应用 | |
| CN116121154B (zh) | 一种乳明串珠菌及其应用 | |
| CN113493491B (zh) | 一种用于预防或治疗溃疡性结肠炎的多肽 | |
| CN113005067A (zh) | 多功能复合益生菌制剂及其制备方法 | |
| CN120310713B (zh) | 长双歧杆菌长亚种及其在生殖系统疾病相关药物中的应用 | |
| US12016361B2 (en) | Probiotic culture and use thereof | |
| KR102380726B1 (ko) | 패칼리박테리움속 미생물 및 이를 포함하는 항암 조성물 | |
| CN117986337B (zh) | 一种提高免疫力的益生菌组合物以及在治疗疾病中的应用 | |
| JP3995733B2 (ja) | 免疫賦活組成物 | |
| CN118383523A (zh) | 凝结芽孢杆菌bc07在制备增强免疫功能食品和/或免疫调节药物和/或抑菌剂方面的用途 | |
| CN117384789A (zh) | 一株戊糖片球菌ks5及其在制备抗氧化和抗衰老食品药品中的应用 | |
| CN113005066B (zh) | 抗过敏、增加免疫力、降血糖、降脂减肥的复合双歧杆菌制剂及其制备方法 | |
| CN111172054B (zh) | 一种干酪乳杆菌组合制剂及其应用 |
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 | ||
| TA01 | Transfer of patent application right | ||
| TA01 | Transfer of patent application right |
Effective date of registration: 20240725 Address after: 310000 Rooms 803 and 805, 8th Floor, Building 1, No. 36 Macheng Road, Xihu District, Hangzhou, Zhejiang Applicant after: HANGZHOU YUSHOU BIOTECHNOLOGY CO.,LTD. Country or region after: China Address before: Unit 28-17-3155, Unit 2, Floors 1 to 2, Building 28, Heng'an Community, Yanqing Town, Yanqing District, Beijing, 100000 Applicant before: Beijing Chuanjin Biotechnology Co.,Ltd. Country or region before: China |
|
| GR01 | Patent grant | ||
| GR01 | Patent grant |