CN113730443B - Application of bacteroides fragilis and extract thereof in preparation of medicine for preventing and treating irritable bowel syndrome - Google Patents
Application of bacteroides fragilis and extract thereof in preparation of medicine for preventing and treating irritable bowel syndrome Download PDFInfo
- Publication number
- CN113730443B CN113730443B CN202110977532.0A CN202110977532A CN113730443B CN 113730443 B CN113730443 B CN 113730443B CN 202110977532 A CN202110977532 A CN 202110977532A CN 113730443 B CN113730443 B CN 113730443B
- Authority
- CN
- China
- Prior art keywords
- bacteroides fragilis
- group
- capsular polysaccharide
- ibs
- day
- 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
- 241000606124 Bacteroides fragilis Species 0.000 title claims abstract description 142
- 208000002551 irritable bowel syndrome Diseases 0.000 title claims abstract description 94
- 239000003814 drug Substances 0.000 title claims abstract description 42
- 239000000284 extract Substances 0.000 title abstract description 37
- 238000002360 preparation method Methods 0.000 title description 15
- 238000004321 preservation Methods 0.000 claims description 3
- 150000004676 glycans Chemical class 0.000 abstract description 94
- 229920001282 polysaccharide Polymers 0.000 abstract description 94
- 239000005017 polysaccharide Substances 0.000 abstract description 94
- 230000000694 effects Effects 0.000 abstract description 39
- 206010012735 Diarrhoea Diseases 0.000 description 64
- 241000700159 Rattus Species 0.000 description 39
- 241000699670 Mus sp. Species 0.000 description 38
- 238000011282 treatment Methods 0.000 description 36
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Substances O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 33
- 206010010774 Constipation Diseases 0.000 description 29
- 210000003608 fece Anatomy 0.000 description 23
- ISWSIDIOOBJBQZ-UHFFFAOYSA-N Phenol Chemical compound OC1=CC=CC=C1 ISWSIDIOOBJBQZ-UHFFFAOYSA-N 0.000 description 22
- 238000002474 experimental method Methods 0.000 description 21
- 229940079593 drug Drugs 0.000 description 20
- 239000002245 particle Substances 0.000 description 20
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 description 19
- 241001465754 Metazoa Species 0.000 description 19
- 230000002550 fecal effect Effects 0.000 description 19
- 239000002244 precipitate Substances 0.000 description 19
- 238000000034 method Methods 0.000 description 16
- 230000002265 prevention Effects 0.000 description 15
- 239000000243 solution Substances 0.000 description 15
- 239000006228 supernatant Substances 0.000 description 14
- 241000699666 Mus <mouse, genus> Species 0.000 description 13
- 238000011740 C57BL/6 mouse Methods 0.000 description 12
- RTZKZFJDLAIYFH-UHFFFAOYSA-N Diethyl ether Chemical compound CCOCC RTZKZFJDLAIYFH-UHFFFAOYSA-N 0.000 description 12
- 238000004458 analytical method Methods 0.000 description 12
- 239000004359 castor oil Substances 0.000 description 12
- 235000019438 castor oil Nutrition 0.000 description 12
- 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 12
- QZAYGJVTTNCVMB-UHFFFAOYSA-N serotonin Chemical compound C1=C(O)C=C2C(CCN)=CNC2=C1 QZAYGJVTTNCVMB-UHFFFAOYSA-N 0.000 description 12
- 210000000813 small intestine Anatomy 0.000 description 12
- 235000019441 ethanol Nutrition 0.000 description 11
- 210000001035 gastrointestinal tract Anatomy 0.000 description 11
- 230000009278 visceral effect Effects 0.000 description 11
- 239000002609 medium Substances 0.000 description 10
- 239000002504 physiological saline solution Substances 0.000 description 10
- 208000024891 symptom Diseases 0.000 description 10
- QTBSBXVTEAMEQO-UHFFFAOYSA-N Acetic acid Chemical compound CC(O)=O QTBSBXVTEAMEQO-UHFFFAOYSA-N 0.000 description 9
- 241000894007 species Species 0.000 description 9
- 230000000968 intestinal effect Effects 0.000 description 8
- 230000002829 reductive effect Effects 0.000 description 8
- 108091005482 5-HT4 receptors Proteins 0.000 description 7
- 208000004998 Abdominal Pain Diseases 0.000 description 7
- 229960002362 neostigmine Drugs 0.000 description 7
- LULNWZDBKTWDGK-UHFFFAOYSA-M neostigmine bromide Chemical compound [Br-].CN(C)C(=O)OC1=CC=CC([N+](C)(C)C)=C1 LULNWZDBKTWDGK-UHFFFAOYSA-M 0.000 description 7
- 230000035945 sensitivity Effects 0.000 description 7
- 238000003756 stirring Methods 0.000 description 7
- 239000000725 suspension Substances 0.000 description 7
- 230000001225 therapeutic effect Effects 0.000 description 7
- 238000005481 NMR spectroscopy Methods 0.000 description 6
- 210000001072 colon Anatomy 0.000 description 6
- 230000000593 degrading effect Effects 0.000 description 6
- 238000013400 design of experiment Methods 0.000 description 6
- 239000010871 livestock manure Substances 0.000 description 6
- RDOIQAHITMMDAJ-UHFFFAOYSA-N loperamide Chemical compound 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 RDOIQAHITMMDAJ-UHFFFAOYSA-N 0.000 description 6
- XQYZDYMELSJDRZ-UHFFFAOYSA-N papaverine Chemical compound C1=C(OC)C(OC)=CC=C1CC1=NC=CC2=CC(OC)=C(OC)C=C12 XQYZDYMELSJDRZ-UHFFFAOYSA-N 0.000 description 6
- 239000006041 probiotic Substances 0.000 description 6
- 235000018291 probiotics Nutrition 0.000 description 6
- 150000003839 salts Chemical class 0.000 description 6
- 241000606125 Bacteroides Species 0.000 description 5
- 235000006693 Cassia laevigata Nutrition 0.000 description 5
- 241000555745 Sciuridae Species 0.000 description 5
- 241000522641 Senna Species 0.000 description 5
- 239000008346 aqueous phase Substances 0.000 description 5
- 239000002775 capsule Substances 0.000 description 5
- 230000008859 change Effects 0.000 description 5
- 238000004108 freeze drying Methods 0.000 description 5
- 230000006870 function Effects 0.000 description 5
- 230000002496 gastric effect Effects 0.000 description 5
- 239000007788 liquid Substances 0.000 description 5
- 230000001376 precipitating effect Effects 0.000 description 5
- 238000001556 precipitation Methods 0.000 description 5
- 230000001737 promoting effect Effects 0.000 description 5
- 239000000018 receptor agonist Substances 0.000 description 5
- 229940044601 receptor agonist Drugs 0.000 description 5
- 229940124513 senna glycoside Drugs 0.000 description 5
- 210000002460 smooth muscle Anatomy 0.000 description 5
- 108091005477 5-HT3 receptors Proteins 0.000 description 4
- 102000035037 5-HT3 receptors Human genes 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 4
- 208000000269 Hyperkinesis Diseases 0.000 description 4
- 229960000583 acetic acid Drugs 0.000 description 4
- 239000007864 aqueous solution Substances 0.000 description 4
- 230000001580 bacterial effect Effects 0.000 description 4
- 230000037396 body weight Effects 0.000 description 4
- 239000000428 dust Substances 0.000 description 4
- 238000000556 factor analysis Methods 0.000 description 4
- 239000008187 granular material Substances 0.000 description 4
- 238000010438 heat treatment Methods 0.000 description 4
- 229960001571 loperamide Drugs 0.000 description 4
- 239000002075 main ingredient Substances 0.000 description 4
- 239000000546 pharmaceutical excipient Substances 0.000 description 4
- 238000011160 research Methods 0.000 description 4
- 230000035807 sensation Effects 0.000 description 4
- 238000007619 statistical method Methods 0.000 description 4
- 239000003826 tablet Substances 0.000 description 4
- 229960002876 tegaserod Drugs 0.000 description 4
- IKBKZGMPCYNSLU-RGVLZGJSSA-N tegaserod Chemical compound C1=C(OC)C=C2C(/C=N/NC(=N)NCCCCC)=CNC2=C1 IKBKZGMPCYNSLU-RGVLZGJSSA-N 0.000 description 4
- 238000012360 testing method Methods 0.000 description 4
- DJHHDLMTUOLVHY-UHFFFAOYSA-N 1,2,3,4-tetrachlorodibenzodioxine Chemical compound C1=CC=C2OC3=C(Cl)C(Cl)=C(Cl)C(Cl)=C3OC2=C1 DJHHDLMTUOLVHY-UHFFFAOYSA-N 0.000 description 3
- LORDFXWUHHSAQU-UHFFFAOYSA-N 3,4,5-trimethoxybenzoic acid [2-(dimethylamino)-2-phenylbutyl] ester Chemical compound C=1C=CC=CC=1C(CC)(N(C)C)COC(=O)C1=CC(OC)=C(OC)C(OC)=C1 LORDFXWUHHSAQU-UHFFFAOYSA-N 0.000 description 3
- 229930008281 A03AD01 - Papaverine Natural products 0.000 description 3
- CSCPPACGZOOCGX-UHFFFAOYSA-N Acetone Chemical compound CC(C)=O CSCPPACGZOOCGX-UHFFFAOYSA-N 0.000 description 3
- 241000186000 Bifidobacterium Species 0.000 description 3
- 241000282412 Homo Species 0.000 description 3
- 206010020651 Hyperkinesia Diseases 0.000 description 3
- IKGXLCMLVINENI-QOXGANSBSA-M [Br-].COc1cc(Br)c(C[N+]2(CCOCC[C@@H]3CC[C@H]4C[C@@H]3C4(C)C)CCOCC2)cc1OC Chemical compound [Br-].COc1cc(Br)c(C[N+]2(CCOCC[C@@H]3CC[C@H]4C[C@@H]3C4(C)C)CCOCC2)cc1OC IKGXLCMLVINENI-QOXGANSBSA-M 0.000 description 3
- OIPILFWXSMYKGL-UHFFFAOYSA-N acetylcholine Chemical compound CC(=O)OCC[N+](C)(C)C OIPILFWXSMYKGL-UHFFFAOYSA-N 0.000 description 3
- 229960004373 acetylcholine Drugs 0.000 description 3
- 239000008280 blood Substances 0.000 description 3
- 210000004369 blood Anatomy 0.000 description 3
- 239000003610 charcoal Substances 0.000 description 3
- 238000010790 dilution Methods 0.000 description 3
- 239000012895 dilution Substances 0.000 description 3
- HYPPXZBJBPSRLK-UHFFFAOYSA-N diphenoxylate Chemical compound C1CC(C(=O)OCC)(C=2C=CC=CC=2)CCN1CCC(C#N)(C=1C=CC=CC=1)C1=CC=CC=C1 HYPPXZBJBPSRLK-UHFFFAOYSA-N 0.000 description 3
- 229960004192 diphenoxylate Drugs 0.000 description 3
- 238000011156 evaluation Methods 0.000 description 3
- 238000000855 fermentation Methods 0.000 description 3
- 230000004151 fermentation Effects 0.000 description 3
- 239000012362 glacial acetic acid Substances 0.000 description 3
- 230000002401 inhibitory effect Effects 0.000 description 3
- 230000005764 inhibitory process Effects 0.000 description 3
- 239000008141 laxative Substances 0.000 description 3
- 230000033001 locomotion Effects 0.000 description 3
- 239000000463 material Substances 0.000 description 3
- 238000009629 microbiological culture Methods 0.000 description 3
- 210000004877 mucosa Anatomy 0.000 description 3
- 229960001789 papaverine Drugs 0.000 description 3
- 230000000529 probiotic effect Effects 0.000 description 3
- 238000011552 rat model Methods 0.000 description 3
- 229940044551 receptor antagonist Drugs 0.000 description 3
- 239000002464 receptor antagonist Substances 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 229940076742 senna leaves Drugs 0.000 description 3
- -1 spasmolytics Substances 0.000 description 3
- 238000001228 spectrum Methods 0.000 description 3
- 239000000126 substance Substances 0.000 description 3
- 229960004354 tegaserod maleate Drugs 0.000 description 3
- 229960005345 trimebutine Drugs 0.000 description 3
- RTHCYVBBDHJXIQ-MRXNPFEDSA-N (R)-fluoxetine Chemical compound O([C@H](CCNC)C=1C=CC=CC=1)C1=CC=C(C(F)(F)F)C=C1 RTHCYVBBDHJXIQ-MRXNPFEDSA-N 0.000 description 2
- 206010000060 Abdominal distension Diseases 0.000 description 2
- 206010067484 Adverse reaction Diseases 0.000 description 2
- 208000019901 Anxiety disease Diseases 0.000 description 2
- 241000423333 Bacteroides fragilis NCTC 9343 Species 0.000 description 2
- 108090000312 Calcium Channels Proteins 0.000 description 2
- 102000003922 Calcium Channels Human genes 0.000 description 2
- BHPQYMZQTOCNFJ-UHFFFAOYSA-N Calcium cation Chemical compound [Ca+2] BHPQYMZQTOCNFJ-UHFFFAOYSA-N 0.000 description 2
- OKTJSMMVPCPJKN-UHFFFAOYSA-N Carbon Chemical compound [C] OKTJSMMVPCPJKN-UHFFFAOYSA-N 0.000 description 2
- 108010009685 Cholinergic Receptors Proteins 0.000 description 2
- 238000003794 Gram staining Methods 0.000 description 2
- 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 description 2
- 230000005856 abnormality Effects 0.000 description 2
- 102000034337 acetylcholine receptors Human genes 0.000 description 2
- 230000009471 action Effects 0.000 description 2
- 230000006838 adverse reaction Effects 0.000 description 2
- 239000000935 antidepressant agent Substances 0.000 description 2
- 229940005513 antidepressants Drugs 0.000 description 2
- 239000003793 antidiarrheal agent Substances 0.000 description 2
- 229940125714 antidiarrheal agent Drugs 0.000 description 2
- 230000036506 anxiety Effects 0.000 description 2
- 239000002249 anxiolytic agent Substances 0.000 description 2
- 230000000949 anxiolytic effect Effects 0.000 description 2
- 230000002457 bidirectional effect Effects 0.000 description 2
- 230000033228 biological regulation Effects 0.000 description 2
- 229910001424 calcium ion Inorganic materials 0.000 description 2
- 229910052799 carbon Inorganic materials 0.000 description 2
- 230000015556 catabolic process Effects 0.000 description 2
- 239000000799 cathartic agent Substances 0.000 description 2
- 238000006243 chemical reaction Methods 0.000 description 2
- 239000003795 chemical substances by application Substances 0.000 description 2
- 239000000812 cholinergic antagonist Substances 0.000 description 2
- 150000001875 compounds Chemical class 0.000 description 2
- 238000011109 contamination Methods 0.000 description 2
- 230000008602 contraction Effects 0.000 description 2
- 238000005100 correlation spectroscopy Methods 0.000 description 2
- 238000012258 culturing Methods 0.000 description 2
- 230000013872 defecation Effects 0.000 description 2
- 238000006731 degradation reaction Methods 0.000 description 2
- GUJOJGAPFQRJSV-UHFFFAOYSA-N dialuminum;dioxosilane;oxygen(2-);hydrate Chemical compound O.[O-2].[O-2].[O-2].[Al+3].[Al+3].O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O GUJOJGAPFQRJSV-UHFFFAOYSA-N 0.000 description 2
- 230000000741 diarrhetic effect Effects 0.000 description 2
- 239000002552 dosage form Substances 0.000 description 2
- 230000000688 enterotoxigenic effect Effects 0.000 description 2
- 230000002964 excitative effect Effects 0.000 description 2
- 239000000194 fatty acid Substances 0.000 description 2
- 229960002464 fluoxetine Drugs 0.000 description 2
- 238000009472 formulation Methods 0.000 description 2
- 210000003736 gastrointestinal content Anatomy 0.000 description 2
- 230000005176 gastrointestinal motility Effects 0.000 description 2
- 238000005227 gel permeation chromatography Methods 0.000 description 2
- 230000036541 health Effects 0.000 description 2
- 238000003919 heteronuclear multiple bond coherence Methods 0.000 description 2
- 208000015181 infectious disease Diseases 0.000 description 2
- 230000003871 intestinal function Effects 0.000 description 2
- 210000004347 intestinal mucosa Anatomy 0.000 description 2
- JVTAAEKCZFNVCJ-UHFFFAOYSA-N lactic acid Chemical compound CC(O)C(O)=O JVTAAEKCZFNVCJ-UHFFFAOYSA-N 0.000 description 2
- 239000008101 lactose Substances 0.000 description 2
- 230000002475 laxative effect Effects 0.000 description 2
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 description 2
- 230000010534 mechanism of action Effects 0.000 description 2
- 238000002156 mixing Methods 0.000 description 2
- 239000000203 mixture Substances 0.000 description 2
- 238000012986 modification Methods 0.000 description 2
- 230000004048 modification Effects 0.000 description 2
- 229910052901 montmorillonite Inorganic materials 0.000 description 2
- 230000000877 morphologic effect Effects 0.000 description 2
- 238000000465 moulding Methods 0.000 description 2
- 208000035824 paresthesia Diseases 0.000 description 2
- 230000008855 peristalsis Effects 0.000 description 2
- 239000006187 pill Substances 0.000 description 2
- 229960002088 pinaverium bromide Drugs 0.000 description 2
- 229920001223 polyethylene glycol Polymers 0.000 description 2
- 230000011514 reflex Effects 0.000 description 2
- 230000002040 relaxant effect Effects 0.000 description 2
- 230000028327 secretion Effects 0.000 description 2
- 210000000329 smooth muscle myocyte Anatomy 0.000 description 2
- 239000000050 smooth muscle relaxant Substances 0.000 description 2
- 238000000527 sonication Methods 0.000 description 2
- 230000002048 spasmolytic effect Effects 0.000 description 2
- 238000010186 staining Methods 0.000 description 2
- 230000000638 stimulation Effects 0.000 description 2
- 238000003860 storage Methods 0.000 description 2
- 210000003934 vacuole Anatomy 0.000 description 2
- 208000009935 visceral pain Diseases 0.000 description 2
- 238000005303 weighing Methods 0.000 description 2
- AHOUBRCZNHFOSL-YOEHRIQHSA-N (+)-Casbol Chemical compound C1=CC(F)=CC=C1[C@H]1[C@H](COC=2C=C3OCOC3=CC=2)CNCC1 AHOUBRCZNHFOSL-YOEHRIQHSA-N 0.000 description 1
- SFLSHLFXELFNJZ-QMMMGPOBSA-N (-)-norepinephrine Chemical compound NC[C@H](O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-QMMMGPOBSA-N 0.000 description 1
- FELGMEQIXOGIFQ-CYBMUJFWSA-N (3r)-9-methyl-3-[(2-methylimidazol-1-yl)methyl]-2,3-dihydro-1h-carbazol-4-one Chemical compound CC1=NC=CN1C[C@@H]1C(=O)C(C=2C(=CC=CC=2)N2C)=C2CC1 FELGMEQIXOGIFQ-CYBMUJFWSA-N 0.000 description 1
- VYVKHNNGDFVQGA-UHFFFAOYSA-N 3,4-dimethoxybenzoic acid 4-[ethyl-[1-(4-methoxyphenyl)propan-2-yl]amino]butyl ester Chemical compound C=1C=C(OC)C=CC=1CC(C)N(CC)CCCCOC(=O)C1=CC=C(OC)C(OC)=C1 VYVKHNNGDFVQGA-UHFFFAOYSA-N 0.000 description 1
- RYYCJUAHISIHTL-UHFFFAOYSA-N 5-azaorotic acid Chemical compound OC(=O)C1=NC(=O)NC(=O)N1 RYYCJUAHISIHTL-UHFFFAOYSA-N 0.000 description 1
- 108091032151 5-hydroxytryptamine receptor family Proteins 0.000 description 1
- 102000040125 5-hydroxytryptamine receptor family Human genes 0.000 description 1
- WTQYWNWRJNXDEG-UHFFFAOYSA-N 6-Hydroxy-hyoscyamin Natural products CN1C(C2)CC(O)C1CC2OC(=O)C(CO)C1=CC=CC=C1 WTQYWNWRJNXDEG-UHFFFAOYSA-N 0.000 description 1
- 108060003345 Adrenergic Receptor Proteins 0.000 description 1
- 102000017910 Adrenergic receptor Human genes 0.000 description 1
- 206010001497 Agitation Diseases 0.000 description 1
- 229920000856 Amylose Polymers 0.000 description 1
- 241001106067 Atropa Species 0.000 description 1
- 229930003347 Atropine Natural products 0.000 description 1
- 241000605059 Bacteroidetes Species 0.000 description 1
- CPELXLSAUQHCOX-UHFFFAOYSA-M Bromide Chemical compound [Br-] CPELXLSAUQHCOX-UHFFFAOYSA-M 0.000 description 1
- OYPRJOBELJOOCE-UHFFFAOYSA-N Calcium Chemical compound [Ca] OYPRJOBELJOOCE-UHFFFAOYSA-N 0.000 description 1
- 229940127291 Calcium channel antagonist Drugs 0.000 description 1
- 108091006146 Channels Proteins 0.000 description 1
- VEXZGXHMUGYJMC-UHFFFAOYSA-M Chloride anion Chemical compound [Cl-] VEXZGXHMUGYJMC-UHFFFAOYSA-M 0.000 description 1
- 208000018672 Dilatation Diseases 0.000 description 1
- 239000004150 EU approved colour Substances 0.000 description 1
- 101710146739 Enterotoxin Proteins 0.000 description 1
- 241000192125 Firmicutes Species 0.000 description 1
- 208000014540 Functional gastrointestinal disease Diseases 0.000 description 1
- 208000018522 Gastrointestinal disease Diseases 0.000 description 1
- 206010052402 Gastrointestinal hypermotility Diseases 0.000 description 1
- 206010017999 Gastrointestinal pain Diseases 0.000 description 1
- 108010010803 Gelatin Proteins 0.000 description 1
- 206010018910 Haemolysis Diseases 0.000 description 1
- RKUNBYITZUJHSG-UHFFFAOYSA-N Hyosciamin-hydrochlorid Natural products CN1C(C2)CCC1CC2OC(=O)C(CO)C1=CC=CC=C1 RKUNBYITZUJHSG-UHFFFAOYSA-N 0.000 description 1
- 206010022678 Intestinal infections Diseases 0.000 description 1
- 108090000862 Ion Channels Proteins 0.000 description 1
- 102000004310 Ion Channels Human genes 0.000 description 1
- 241000186660 Lactobacillus Species 0.000 description 1
- 229940121948 Muscarinic receptor antagonist Drugs 0.000 description 1
- 102000003840 Opioid Receptors Human genes 0.000 description 1
- 108090000137 Opioid Receptors Proteins 0.000 description 1
- 108070000021 Opioid peptides receptors Proteins 0.000 description 1
- AHOUBRCZNHFOSL-UHFFFAOYSA-N Paroxetine hydrochloride Natural products C1=CC(F)=CC=C1C1C(COC=2C=C3OCOC3=CC=2)CNCC1 AHOUBRCZNHFOSL-UHFFFAOYSA-N 0.000 description 1
- 239000004264 Petrolatum Substances 0.000 description 1
- 239000002202 Polyethylene glycol Substances 0.000 description 1
- 102000004257 Potassium Channel Human genes 0.000 description 1
- DNIAPMSPPWPWGF-UHFFFAOYSA-N Propylene glycol Chemical class CC(O)CO DNIAPMSPPWPWGF-UHFFFAOYSA-N 0.000 description 1
- 208000035755 Psychosomatic disease Diseases 0.000 description 1
- 208000028017 Psychotic disease Diseases 0.000 description 1
- 108010012996 Serotonin Plasma Membrane Transport Proteins Proteins 0.000 description 1
- 102000019208 Serotonin Plasma Membrane Transport Proteins Human genes 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 229920002472 Starch Polymers 0.000 description 1
- 229940123445 Tricyclic antidepressant Drugs 0.000 description 1
- 208000027418 Wounds and injury Diseases 0.000 description 1
- 210000001015 abdomen Anatomy 0.000 description 1
- 210000000683 abdominal cavity Anatomy 0.000 description 1
- 206010060926 abdominal symptom Diseases 0.000 description 1
- 206010000269 abscess Diseases 0.000 description 1
- 238000010521 absorption reaction Methods 0.000 description 1
- 238000009825 accumulation Methods 0.000 description 1
- 102000019997 adhesion receptor Human genes 0.000 description 1
- 108010013985 adhesion receptor Proteins 0.000 description 1
- 239000000443 aerosol Substances 0.000 description 1
- 150000001298 alcohols Chemical class 0.000 description 1
- FLZQKRKHLSUHOR-UHFFFAOYSA-N alosetron Chemical compound CC1=NC=N[C]1CN1C(=O)C(C=2C(=CC=CC=2)N2C)=C2CC1 FLZQKRKHLSUHOR-UHFFFAOYSA-N 0.000 description 1
- 229960003550 alosetron Drugs 0.000 description 1
- 229960000836 amitriptyline Drugs 0.000 description 1
- KRMDCWKBEZIMAB-UHFFFAOYSA-N amitriptyline Chemical compound C1CC2=CC=CC=C2C(=CCCN(C)C)C2=CC=CC=C21 KRMDCWKBEZIMAB-UHFFFAOYSA-N 0.000 description 1
- 230000000202 analgesic effect Effects 0.000 description 1
- WTQYWNWRJNXDEG-LEOABGAYSA-N anisodamine Chemical compound C1([C@@H](CO)C(=O)O[C@@H]2C[C@H]3[C@@H](O)C[C@@H](C2)N3C)=CC=CC=C1 WTQYWNWRJNXDEG-LEOABGAYSA-N 0.000 description 1
- 239000005557 antagonist Substances 0.000 description 1
- 230000001078 anti-cholinergic effect Effects 0.000 description 1
- 230000001430 anti-depressive effect Effects 0.000 description 1
- 230000001142 anti-diarrhea Effects 0.000 description 1
- 238000013459 approach Methods 0.000 description 1
- RKUNBYITZUJHSG-SPUOUPEWSA-N atropine Chemical compound O([C@H]1C[C@H]2CC[C@@H](C1)N2C)C(=O)C(CO)C1=CC=CC=C1 RKUNBYITZUJHSG-SPUOUPEWSA-N 0.000 description 1
- 229960000396 atropine Drugs 0.000 description 1
- 244000052616 bacterial pathogen Species 0.000 description 1
- 230000002146 bilateral effect Effects 0.000 description 1
- 239000011230 binding agent Substances 0.000 description 1
- 238000006065 biodegradation reaction Methods 0.000 description 1
- 230000004071 biological effect Effects 0.000 description 1
- 230000005540 biological transmission Effects 0.000 description 1
- 230000015572 biosynthetic process Effects 0.000 description 1
- 230000017531 blood circulation Effects 0.000 description 1
- 210000001185 bone marrow Anatomy 0.000 description 1
- 210000005013 brain tissue Anatomy 0.000 description 1
- 239000011575 calcium Substances 0.000 description 1
- 229910052791 calcium Inorganic materials 0.000 description 1
- 239000000480 calcium channel blocker Substances 0.000 description 1
- 150000001720 carbohydrates Chemical class 0.000 description 1
- 210000004027 cell Anatomy 0.000 description 1
- 210000000170 cell membrane Anatomy 0.000 description 1
- 238000002144 chemical decomposition reaction Methods 0.000 description 1
- 230000001713 cholinergic effect Effects 0.000 description 1
- DCSUBABJRXZOMT-IRLDBZIGSA-N cisapride Chemical compound C([C@@H]([C@@H](CC1)NC(=O)C=2C(=CC(N)=C(Cl)C=2)OC)OC)N1CCCOC1=CC=C(F)C=C1 DCSUBABJRXZOMT-IRLDBZIGSA-N 0.000 description 1
- 229960005132 cisapride Drugs 0.000 description 1
- DCSUBABJRXZOMT-UHFFFAOYSA-N cisapride Natural products C1CC(NC(=O)C=2C(=CC(N)=C(Cl)C=2)OC)C(OC)CN1CCCOC1=CC=C(F)C=C1 DCSUBABJRXZOMT-UHFFFAOYSA-N 0.000 description 1
- 239000011248 coating agent Substances 0.000 description 1
- 230000000112 colonic effect Effects 0.000 description 1
- 238000010276 construction Methods 0.000 description 1
- 239000006071 cream Substances 0.000 description 1
- 230000006378 damage Effects 0.000 description 1
- 239000008367 deionised water Substances 0.000 description 1
- 229910021641 deionized water Inorganic materials 0.000 description 1
- 239000007933 dermal patch Substances 0.000 description 1
- 238000000502 dialysis Methods 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000037213 diet Effects 0.000 description 1
- 235000014113 dietary fatty acids Nutrition 0.000 description 1
- 239000003085 diluting agent Substances 0.000 description 1
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 1
- 208000035475 disorder Diseases 0.000 description 1
- 238000009826 distribution Methods 0.000 description 1
- 238000010410 dusting Methods 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 230000000214 effect on organisms Effects 0.000 description 1
- 230000008451 emotion Effects 0.000 description 1
- 210000000105 enteric nervous system Anatomy 0.000 description 1
- 210000005216 enteric neuron Anatomy 0.000 description 1
- 239000000147 enterotoxin Substances 0.000 description 1
- 231100000655 enterotoxin Toxicity 0.000 description 1
- 229930195729 fatty acid Natural products 0.000 description 1
- 239000000796 flavoring agent Substances 0.000 description 1
- 235000013305 food Nutrition 0.000 description 1
- 238000003304 gavage Methods 0.000 description 1
- 239000008273 gelatin Substances 0.000 description 1
- 229920000159 gelatin Polymers 0.000 description 1
- 235000019322 gelatine Nutrition 0.000 description 1
- 235000011852 gelatine desserts Nutrition 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 239000001963 growth medium Substances 0.000 description 1
- 230000010243 gut motility Effects 0.000 description 1
- 230000008588 hemolysis Effects 0.000 description 1
- 241000411851 herbal medicine Species 0.000 description 1
- 230000010196 hermaphroditism Effects 0.000 description 1
- 238000005570 heteronuclear single quantum coherence Methods 0.000 description 1
- 238000000990 heteronuclear single quantum coherence spectrum Methods 0.000 description 1
- 229910052739 hydrogen Inorganic materials 0.000 description 1
- 239000001257 hydrogen Substances 0.000 description 1
- 229920003063 hydroxymethyl cellulose Polymers 0.000 description 1
- 229940031574 hydroxymethyl cellulose Drugs 0.000 description 1
- 230000001660 hyperkinetic effect Effects 0.000 description 1
- 230000035873 hypermotility Effects 0.000 description 1
- 230000006872 improvement Effects 0.000 description 1
- 239000003112 inhibitor Substances 0.000 description 1
- 238000011221 initial treatment Methods 0.000 description 1
- 238000002347 injection Methods 0.000 description 1
- 239000007924 injection Substances 0.000 description 1
- 208000014674 injury Diseases 0.000 description 1
- 208000018936 intestinal hypermotility Diseases 0.000 description 1
- 230000037036 intestinal hypermotility Effects 0.000 description 1
- 210000000936 intestine Anatomy 0.000 description 1
- 238000010255 intramuscular injection Methods 0.000 description 1
- 239000007927 intramuscular injection Substances 0.000 description 1
- 238000010253 intravenous injection Methods 0.000 description 1
- 229920000831 ionic polymer Polymers 0.000 description 1
- 150000002500 ions Chemical class 0.000 description 1
- 239000004310 lactic acid Substances 0.000 description 1
- 235000014655 lactic acid Nutrition 0.000 description 1
- 229940039696 lactobacillus Drugs 0.000 description 1
- JCQLYHFGKNRPGE-FCVZTGTOSA-N lactulose Chemical compound OC[C@H]1O[C@](O)(CO)[C@@H](O)[C@@H]1O[C@H]1[C@H](O)[C@@H](O)[C@@H](O)[C@@H](CO)O1 JCQLYHFGKNRPGE-FCVZTGTOSA-N 0.000 description 1
- 229960000511 lactulose Drugs 0.000 description 1
- PFCRQPBOOFTZGQ-UHFFFAOYSA-N lactulose keto form Natural products OCC(=O)C(O)C(C(O)CO)OC1OC(CO)C(O)C(O)C1O PFCRQPBOOFTZGQ-UHFFFAOYSA-N 0.000 description 1
- 229940125722 laxative agent Drugs 0.000 description 1
- 230000000670 limiting effect Effects 0.000 description 1
- 239000002502 liposome Substances 0.000 description 1
- 210000004185 liver Anatomy 0.000 description 1
- 230000007774 longterm Effects 0.000 description 1
- 239000006210 lotion Substances 0.000 description 1
- 239000007937 lozenge Substances 0.000 description 1
- 239000000314 lubricant Substances 0.000 description 1
- 210000004072 lung Anatomy 0.000 description 1
- 235000019359 magnesium stearate Nutrition 0.000 description 1
- 230000014759 maintenance of location Effects 0.000 description 1
- 229960003577 mebeverine Drugs 0.000 description 1
- 230000028161 membrane depolarization Effects 0.000 description 1
- 230000000813 microbial effect Effects 0.000 description 1
- 150000002772 monosaccharides Chemical class 0.000 description 1
- 230000004899 motility Effects 0.000 description 1
- 210000004400 mucous membrane Anatomy 0.000 description 1
- 210000003205 muscle Anatomy 0.000 description 1
- 239000007922 nasal spray Substances 0.000 description 1
- 229940097496 nasal spray Drugs 0.000 description 1
- 210000005036 nerve Anatomy 0.000 description 1
- 210000002569 neuron Anatomy 0.000 description 1
- 239000002858 neurotransmitter agent Substances 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 229960002748 norepinephrine Drugs 0.000 description 1
- SFLSHLFXELFNJZ-UHFFFAOYSA-N norepinephrine Natural products NCC(O)C1=CC=C(O)C(O)=C1 SFLSHLFXELFNJZ-UHFFFAOYSA-N 0.000 description 1
- 235000016709 nutrition Nutrition 0.000 description 1
- 239000003921 oil Substances 0.000 description 1
- 235000019198 oils Nutrition 0.000 description 1
- 239000002674 ointment Substances 0.000 description 1
- 229960005343 ondansetron Drugs 0.000 description 1
- 244000039328 opportunistic pathogen Species 0.000 description 1
- 229940006093 opthalmologic coloring agent diagnostic Drugs 0.000 description 1
- 210000000056 organ Anatomy 0.000 description 1
- 230000003204 osmotic effect Effects 0.000 description 1
- 229950000193 oteracil Drugs 0.000 description 1
- 230000037040 pain threshold Effects 0.000 description 1
- 239000012188 paraffin wax Substances 0.000 description 1
- 238000007911 parenteral administration Methods 0.000 description 1
- 229960002296 paroxetine Drugs 0.000 description 1
- 230000008506 pathogenesis Effects 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000002304 perfume Substances 0.000 description 1
- 229940066842 petrolatum Drugs 0.000 description 1
- 235000019271 petrolatum Nutrition 0.000 description 1
- 230000000144 pharmacologic effect Effects 0.000 description 1
- 238000011458 pharmacological treatment Methods 0.000 description 1
- 239000012071 phase Substances 0.000 description 1
- 229920001296 polysiloxane Polymers 0.000 description 1
- 239000001267 polyvinylpyrrolidone Substances 0.000 description 1
- 229920000036 polyvinylpyrrolidone Polymers 0.000 description 1
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 description 1
- 239000013641 positive control Substances 0.000 description 1
- 230000000291 postprandial effect Effects 0.000 description 1
- 108020001213 potassium channel Proteins 0.000 description 1
- 239000000843 powder Substances 0.000 description 1
- 230000003449 preventive effect Effects 0.000 description 1
- 230000008569 process Effects 0.000 description 1
- 239000000047 product Substances 0.000 description 1
- 239000002325 prokinetic agent Substances 0.000 description 1
- 235000013772 propylene glycol Nutrition 0.000 description 1
- 150000003180 prostaglandins Chemical class 0.000 description 1
- 230000001681 protective effect Effects 0.000 description 1
- 238000000425 proton nuclear magnetic resonance spectrum Methods 0.000 description 1
- 208000020016 psychiatric disease Diseases 0.000 description 1
- 102000005962 receptors Human genes 0.000 description 1
- 108020003175 receptors Proteins 0.000 description 1
- 230000008439 repair process Effects 0.000 description 1
- 230000000241 respiratory effect Effects 0.000 description 1
- 230000033764 rhythmic process Effects 0.000 description 1
- 239000012266 salt solution Substances 0.000 description 1
- 210000002265 sensory receptor cell Anatomy 0.000 description 1
- 208000026775 severe diarrhea Diseases 0.000 description 1
- RMAQACBXLXPBSY-UHFFFAOYSA-N silicic acid Chemical compound O[Si](O)(O)O RMAQACBXLXPBSY-UHFFFAOYSA-N 0.000 description 1
- 235000012239 silicon dioxide Nutrition 0.000 description 1
- 210000004872 soft tissue Anatomy 0.000 description 1
- 239000007787 solid Substances 0.000 description 1
- 239000002904 solvent Substances 0.000 description 1
- 239000008107 starch Substances 0.000 description 1
- 235000019698 starch Nutrition 0.000 description 1
- 239000008143 stimulant laxative Substances 0.000 description 1
- 238000010254 subcutaneous injection Methods 0.000 description 1
- 239000007929 subcutaneous injection Substances 0.000 description 1
- 230000009469 supplementation Effects 0.000 description 1
- 239000000829 suppository Substances 0.000 description 1
- 239000004094 surface-active agent Substances 0.000 description 1
- 239000000375 suspending agent Substances 0.000 description 1
- 230000008961 swelling Effects 0.000 description 1
- 208000011580 syndromic disease Diseases 0.000 description 1
- 238000003786 synthesis reaction Methods 0.000 description 1
- 239000000454 talc Substances 0.000 description 1
- 229910052623 talc Inorganic materials 0.000 description 1
- 210000004876 tela submucosa Anatomy 0.000 description 1
- 238000002560 therapeutic procedure Methods 0.000 description 1
- 238000011200 topical administration Methods 0.000 description 1
- 230000001988 toxicity Effects 0.000 description 1
- 231100000419 toxicity Toxicity 0.000 description 1
- 229940126680 traditional chinese medicines Drugs 0.000 description 1
- 239000003029 tricyclic antidepressant agent Substances 0.000 description 1
- 239000012137 tryptone Substances 0.000 description 1
- 229910021642 ultra pure water Inorganic materials 0.000 description 1
- 239000012498 ultrapure water Substances 0.000 description 1
- 241001148471 unidentified anaerobic bacterium Species 0.000 description 1
- 235000015112 vegetable and seed oil Nutrition 0.000 description 1
- 239000008158 vegetable oil Substances 0.000 description 1
- 210000001835 viscera Anatomy 0.000 description 1
- 239000001993 wax Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C19/00—Cheese; Cheese preparations; Making thereof
- A23C19/06—Treating cheese curd after whey separation; Products obtained thereby
- A23C19/09—Other cheese preparations; Mixtures of cheese with other foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23C—DAIRY PRODUCTS, e.g. MILK, BUTTER OR CHEESE; MILK OR CHEESE SUBSTITUTES; MAKING THEREOF
- A23C9/00—Milk preparations; Milk powder or milk powder preparations
- A23C9/12—Fermented milk preparations; Treatment using microorganisms or enzymes
- A23C9/13—Fermented milk preparations; Treatment using microorganisms or enzymes using additives
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23G—COCOA; COCOA PRODUCTS, e.g. CHOCOLATE; SUBSTITUTES FOR COCOA OR COCOA PRODUCTS; CONFECTIONERY; CHEWING GUM; ICE-CREAM; PREPARATION THEREOF
- A23G9/00—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor
- A23G9/32—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds
- A23G9/36—Frozen sweets, e.g. ice confectionery, ice-cream; Mixtures therefor characterised by the composition containing organic or inorganic compounds containing microorganisms or enzymes; containing paramedical or dietetical agents, e.g. vitamins
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K10/00—Animal feeding-stuffs
- A23K10/10—Animal feeding-stuffs obtained by microbiological or biochemical processes
- A23K10/16—Addition of microorganisms or extracts thereof, e.g. single-cell proteins, to feeding-stuff compositions
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23K—FODDER
- A23K20/00—Accessory food factors for animal feeding-stuffs
- A23K20/10—Organic substances
- A23K20/163—Sugars; Polysaccharides
-
- 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
- A23L33/125—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives containing carbohydrate syrups; containing sugars; containing sugar alcohols; containing starch hydrolysates
-
- 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
- A61K31/00—Medicinal preparations containing organic active ingredients
- A61K31/70—Carbohydrates; Sugars; Derivatives thereof
- A61K31/715—Polysaccharides, i.e. having more than five saccharide radicals attached to each other by glycosidic linkages; Derivatives thereof, e.g. ethers, esters
-
- 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
-
- 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
-
- 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
- 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
-
- 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
-
- 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
- Life Sciences & Earth Sciences (AREA)
- Health & Medical Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Microbiology (AREA)
- Veterinary Medicine (AREA)
- Pharmacology & Pharmacy (AREA)
- Polymers & Plastics (AREA)
- Animal Behavior & Ethology (AREA)
- Public Health (AREA)
- Mycology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- Organic Chemistry (AREA)
- Food Science & Technology (AREA)
- Molecular Biology (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- General Chemical & Material Sciences (AREA)
- Chemical Kinetics & Catalysis (AREA)
- Epidemiology (AREA)
- Zoology (AREA)
- Nutrition Science (AREA)
- Biotechnology (AREA)
- Biochemistry (AREA)
- Wood Science & Technology (AREA)
- Genetics & Genomics (AREA)
- Biomedical Technology (AREA)
- Animal Husbandry (AREA)
- Tropical Medicine & Parasitology (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Inorganic Chemistry (AREA)
- Physiology (AREA)
- Pharmaceuticals Containing Other Organic And Inorganic Compounds (AREA)
- Medicines Containing Plant Substances (AREA)
- Polysaccharides And Polysaccharide Derivatives (AREA)
Abstract
The invention relates to application of bacteroides fragilis and an extract thereof in preparing a medicament for preventing and treating irritable bowel syndrome, wherein the bacteroides fragilis extract contains bacteroides fragilis capsular polysaccharide A. The invention unexpectedly prepares the bacteroides fragilis capsular polysaccharide A with the molecular weight of 5-70KD, and unexpectedly finds that the capsular polysaccharide A with the molecular weight of 5-70KD has better function of preventing and treating irritable bowel syndrome, and the effect of the bacteroides fragilis capsular polysaccharide A is far better than that of the bacteroides fragilis capsular polysaccharide A with the molecular weight of 110KD extracted from NCTC 9343.
Description
The application is a divisional application of Chinese patent application 201710812676.4, which is filed on 9, 11 and 2017 and is named as application of a bacteroides fragilis extract in preparation of medicines or foods for preventing and treating irritable bowel syndrome.
The microbial strains used in the implementation process of the invention are preserved in China general microbiological culture Collection center (CGMCC) (No. 3 Hospital No.1 Xilu Beijing, chaoyang, beijing) at 4.M.2.2015.4.M.. And (3) classification and naming: bacteroides fragilis ZY-312 (bacteriodes fragilis ZY-312), accession number CGMCC No.10685. Bacteroides fragilis ZY-312 was isolated by the applicant and has been patented (patent No. 201510459408. X), as specified in the patent examination manual, commercially available or licensed to the public without storage, i.e., without providing proof of storage.
Technical Field
The invention relates to the technical field of application of bacteroides fragilis, in particular to application of a bacteroides fragilis extract in preparation of a medicine for preventing and treating irritable bowel syndrome.
Background
Irritable Bowel Syndrome (IBS) is the most common disorder of bowel function in clinic, and has been recognized in recent years as a group of psychosomatic diseases with special pathological and physiological bases, and is a group of syndromes manifested by abdominal pain, abdominal distension, constipation and diarrhea, or constipation and diarrhea alternating and lacking morphological or biochemical abnormalities. IBS has a high global incidence rate, and the incidence rate of IBS is between 10 and 15 percent in every region. In western countries, IBS accounts for 12% of the outpatient services of home health care physicians and 20-50% of the outpatient services of gastrointestinal diseases. The rate of IBS symptoms in China is the same as that in foreign countries, patients mainly take young people and middle-aged people, the age is between 20 and 50 years, and the first-onset patients above 50 years are rare. Women are more than men, and the proportion of men and women is 1 to 5 to 1, and the family tends to aggregate. Studies have shown that the medical resources spent in IBS are considerable, affecting the quality of life of patients to varying degrees.
The etiology and pathogenesis of IBS are still unclear, and it may be related to diet, intestinal infection and psychology, etc., as a result of a combination of factors. The clinical symptoms are various, and besides abdominal symptoms, other symptoms are accompanied, so that the treatment is difficult, and the combination of medicines is often needed. Some patients also have psychopsychological problems such as depression and anxiety, and usually need psychological treatment. Therefore, comprehensive treatment should be applied to treat IBS according to the severity of symptoms, type of symptoms and frequency of attacks of patients, and according to individual treatment principles. The choice of treatment and drug will vary from person to person and include: primary treatment, psychological treatment and drug treatment. At present, the following drugs are mainly used for treating IBS:
1. drugs that modulate intestinal function include antidiarrheals, spasmolytics, prokinetic agents, and agents that modulate visceral sensitivity.
Antidiarrheal agents are commonly used in the treatment of diarrhea in patients with IBS. Commonly used drugs are loperamide, diphenoxylate and dioctahedral montmorillonite. Loperamide (Yimanting) acts on opioid peptide receptors on intestinal walls, prevents acetylcholine and prostaglandin from releasing, inhibits intestinal peristalsis, prolongs the retention time of intestinal contents, enhances the absorption of water and ions in intestinal tracts, and further relieves diarrhea, abdominal pain and the like. The diphenoxylate (diphenoxylate) acts on intestinal smooth muscle, increases segmental contraction of the intestine and prolongs the contact time of intestinal contents and intestinal mucosa. The dioctahedral montmorillonite (Cimpanida) can absorb water and pathogenic bacteria, improve the protective power of digestive tract mucosa, promote mucosa repair, and simultaneously can regulate and recover colon motion function and reduce colon sensitivity.
Spasmolytic is commonly used for abdominal pain and abdominal distension in patients with IBS. They can be classified into 3 groups according to their main mechanism of action, namely anticholinergic drugs, smooth muscle relaxants and calcium channel blockers, many of which have multiple pharmacological actions.
Anticholinergic agents include atropine, anisodamine, belladonna, etc. It has atropine-like adverse reaction, so it has limited clinical application. Recently developed gut M3 selective cholinergic receptor antagonists may inhibit postprandial gut motility and are expected to be useful in the treatment of IBS.
Smooth muscle relaxants include papaverine drugs (papaverine, bicyclovine, mebeverine) and polyion channel modulators (trimebutine). The papaverine medicine can directly act on smooth muscle cells and certain intestinal tract excitatory neurons to inhibit the release of excitatory neurotransmitters. The trimebutine can inhibit the potassium ion channel of the cell membrane to generate depolarization, thereby improving the excitability of smooth muscle cells; on the other hand, the calcium ion channel is blocked to inhibit the calcium ion inflow, thereby inhibiting the cell contraction and relaxing the smooth muscle of the gastrointestinal tract. In addition, trimebutine also has a bidirectional modulating effect on smooth muscle neuroreceptors: under the condition of low motion, the compound acts on adrenergic receptors, inhibits the release of norepinephrine and increases the motor rhythm; when hyperkinesia occurs, the compound acts on cholinergic receptors and opioid receptors to inhibit acetylcholine release, thereby inhibiting smooth muscle movement.
The calcium ion antagonist selectively acts on colon calcium ion channel, blocks calcium inflow, plays a role in relaxing smooth muscle, inhibiting gastric and colonic reflex, has a regulating effect on constipation and diarrhea, and also has a certain curative effect on abdominal pain, such as pinaverium bromide and oteracil bromide.
Prokinetic drugs are commonly used for the treatment of constipation in patients with IBS. 5-hydroxytryptamine (5-HT) is an important transmitter in the gastrointestinal tract and the center, has wide biological effects, 95 percent of 5-HT in human bodies is distributed in the gastrointestinal tract, and various 5-HT receptors and 5-HT transporters are distributed on intestinal mucosa. In recent years, the important role of 5-HT4 receptor in the regulation of gastrointestinal motility and visceral sensation has been emphasized, and it has the effects of promoting gastrointestinal motility, reducing gastrointestinal sensitivity, and promoting the secretion of chloride ions and water molecules, thus becoming a new target for the treatment of functional gastrointestinal diseases. The 5-HT4 receptor agonist cisapride can promote the release of acetylcholine from the postganglionic cholinergic nerve of the myenteric muscle, and has the function of promoting the gastrointestinal tract. However, it should be used with cautions because it may cause prolongation of QT interval. Tegaserod, also known as zerumab, is a novel 5-HT4 receptor agonist, partially selectively acts on gastrointestinal 5-HT4 receptor subtypes, and has the effect of accelerating small intestine and colon transmission in constipation-predominant IBS patients. Recent research also proves that tegaserod has a regulating effect on internal organ sensation, has no cardiovascular adverse reaction, and is a safe and effective novel medicine.
Agents that modulate visceral sensitivity include 5-HT3 receptor antagonists and 5-HT4 receptor agonists. Increased visceral sensitivity is considered to be one of the pathologically and physiologically important features of IBS. Studies have shown that visceral paresthesia is present in 61% of IBS patients, and improving visceral paresthesia is an interesting approach in IBS treatment, and clinical and animal experimental studies have shown that some drugs have a modulating effect on increased visceral sensitivity.
The 5-HT3 receptors are present in enteric neurons and promote motility, secretion and visceral pain stimulation of the gut by local release of 5-HT. For patients with IBS, especially those with abdominal pain due to a decrease in visceral pain threshold, 5-HT3 receptor antagonists are being tried. Alosetron, for example, mainly inhibits the 5-HT3 receptor of a non-selective ion channel in the enteric nervous system and inhibits the visceral reflex, and is recently mainly used for female IBS patients with severe diarrhea who are not treated conventionally. Still others are ondansetron, gonacetone, and the like.
The 5-HT4 receptor agonist tegaserod has the double effects of promoting force and reducing visceral sensation sensitivity, and is suitable for constipation type IBS patients with obvious abdominal pain symptoms. Human body research reports that tegaserod can reduce the reaction of rectal balloon dilatation injury stimulation and improve the visceral sensation of a human body.
2. A cathartic agent: in addition to 5-HT4 receptor agonists, cathartics may also be used in patients with constipation. It is presently believed that the use of stimulant laxatives should be cautious or avoided as much as possible, and the use of bulking laxatives is recommended. The swelling laxative such as herba plantaginis can increase the volume and water content of feces and promote defecation. Osmotic laxative polyethylene glycol binds water molecules through hydrogen bonds, increasing fecal water content and softening feces, promoting defecation. It is not absorbed, and has little toxicity, and is suitable for patients with dry stool. Lactulose is decomposed by bacteria in the colon to form lactic acid and acetic acid, which regulate the environment in the intestinal lumen and promote intestinal peristalsis, and is more suitable for the elderly.
3. Drugs that improve central emotion: patients with IBS are often associated with psychopsychological disturbances, and pharmacological treatment for IBS should include antidepressant, anxiolytic treatment. Antidepressants include tricyclic antidepressants, such as amitriptyline, and selective 5-HT reuptake inhibitors, such as fluoxetine and paroxetine. Wherein, fluoxetine has 4 effects on treating IBS in that psychopsychological improvement induces relief of gastrointestinal symptoms; 5-HT transmitter activity and its receptor action on gastrointestinal tract power for regulating visceral gastrointestinal pain; potential central analgesic effects; preventing the vicious circle between psychological disorders and physiological abnormalities. For IBS patients with severe anxiety psychotic symptoms, anxiolytic therapy may be considered in the treatment.
4. The traditional Chinese medicine comprises the following components: some clinical observations in China prove that some traditional Chinese medicines can effectively relieve the symptoms of abdominal pain, diarrhea and constipation of IBS patients.
5. A microecological preparation: in recent years, attention has been paid to the relationship between intestinal flora disturbance and IBS. Studies have shown that the incidence of a subset of patients is associated with a dysregulated intestinal flora, and that the supplementation of the patients with probiotics improves the symptoms of the patients to varying degrees. The mechanism of action is not well understood, and biochemical inhibition or promotion, nutritional competition, immune clearance, and adhesion receptor competition are generally considered. However, the types of probiotic preparations commonly used are not limited, and mainly include live bifidobacterium triple, live bifidobacterium quadruple and the like.
Currently, there is no very effective treatment for IBS and there is no complete cure for IBS. There is a need to develop new effective drugs that can treat IBS.
Bacteroides fragilis (Bacteroides fragilis) is a member of the genus Bacteroides among gram-negative anaerobic bacteria, belongs to the phylum bacteroidetes, and is completely different from bifidobacterium, lactobacillus, and the like of the phylum firmicutes. Bacteroides have 25 species, only 10 species from humans, only 10 species from animals, and 5 species from humans and animals. The bacteroides fragilis is an obligate anaerobic bacterium, the shape of the bacteroides is polymorphic according to the difference of culture media and the difference of growth stages, the bacteroides fragilis is rod-shaped, the two ends of the bacteroides fragilis are blunt and round, the coloration is dark, the middle color is light and uneven, the bacteroides fragilis, no spores and no power exist, some bacteroides have vacuoles, and the bacteroides are different in length. Bacteriodes fragilis enterotoxin (BFT) can be classified into Enterotoxigenic bacteriodes fragilis (ETBF) and non-Enterotoxigenic bacteriodes fragilis (NTBF) depending on whether they can be synthesized and secreted. Bacteroides fragilis is found predominantly in the colon as part of the normal intestinal flora of humans and animals. In addition, mucous membranes of the respiratory, gastrointestinal and genitourinary tracts may colonize. Bacteroides fragilis, a opportunistic pathogen, when damaged in the host mucosa can invade the submucosa and cause infection, and can also cause purulent infection and abscess associated with it in other organs of the body, such as the intestinal tract, abdominal cavity, liver, lung, brain tissue, soft tissue, bone marrow, etc., by blood flow.
Disclosure of Invention
Based on this, the present invention provides a novel use of Bacteroides fragilis (Bacteroides fragilis) extract. The specific technical scheme is as follows:
application of a bacteroides fragilis extract in preparation of a medicine for preventing and treating irritable bowel syndrome, wherein the bacteroides fragilis extract contains bacteroides fragilis capsular polysaccharide A.
In some embodiments, the bacteroides fragilis capsular polysaccharide a has a molecular weight of 5-75 KD.
In some embodiments, the bacteroides fragilis capsular polysaccharide A has a molecular weight of 15 KD-65 KD;
in some embodiments, the bacteroides fragilis capsular polysaccharide A has a molecular weight of 25 KD-55K.
In some embodiments, the bacteroides fragilis capsular polysaccharide a has a molecular weight of 35KD to 45KD.
In some of these embodiments, the bacteroides fragilis extract contains bacteroides fragilis capsular polysaccharide a in an amount of 60-75wt%.
In some embodiments, the bacteroides fragilis is bacteroides fragilis ZY-312 with accession number of CGMCC No.10685.
In some of these embodiments, the method of preparing the bacteroides fragilis extract comprises the steps of:
(1) Centrifuging and precipitating the bacteroides fragilis bacterial liquid after fermentation culture, collecting a first precipitate, adding water with the temperature of 65-72 ℃ into the first precipitate, adding a phenol solution after dissolving, keeping the temperature of 65-72 ℃, stirring for 25-35min, centrifuging, and collecting a first supernatant;
(2) Extracting the first supernatant collected in the step (1) by using ether to remove phenol, removing residual ether, and collecting an aqueous phase solution;
(3) Adding absolute ethyl alcohol into the aqueous phase solution collected in the step (2) until the final concentration of the ethyl alcohol is 75-85v/v%, precipitating with alcohol, centrifuging, collecting a second precipitate,
(4) And adding water into the second precipitate to prepare a suspension, adjusting the pH to 6.5-7.5, centrifuging, collecting a second supernatant, dialyzing to remove salt, and freeze-drying to obtain the bacteroides fragilis extract.
In some embodiments, the ratio of the water added to the first precipitate in step (1), the phenol solution and the first precipitate is 3-5mL:3-5mL:1g; the mass concentration of the phenol solution is 70-80%.
In some embodiments, the alcohol precipitation in step (3) is alcohol precipitation at a temperature of 0-8 ℃ for 8-16 hours.
In some embodiments, step (4) comprises: and adding water into the second precipitate to prepare a suspension with the mass concentration of 8-12%, adding a glacial acetic acid aqueous solution with the mass concentration of 8-12%, heating to boil, stirring for reaction for 1.5-2.5 hours, adjusting the pH value to 6.5-7.5, centrifuging, collecting a second supernatant, dialyzing to remove salt, and freeze-drying to obtain the bacteroides fragilis extract.
In some of these embodiments, the method of preparing the bacteroides fragilis extract further comprises the step of degrading: degrading the bacteroides fragilis extract obtained in the step (4) by using an ultrasonic method, wherein the ultrasonic condition is as follows: 180-210kHz,15-25 ℃.
In some embodiments, the dosage form of the medicament comprises a pill, tablet, granule, capsule, oral liquid, or tube feed formulation. The medicine comprises human medicine or animal medicine, and can be used for human or animal.
The bacteroides fragilis extract can be administered prophylactically or therapeutically alone, or with other probiotics and/or probiotic materials. In the case of combined administration, the administration may be carried out in a single preparation or in separate preparations, simultaneously or at different times, using the same or different routes of administration.
The invention also provides a medicine for preventing and treating irritable bowel syndrome. The specific technical scheme is as follows:
a medicine for preventing and treating irritable bowel syndrome contains a bacteroides fragilis extract, wherein the bacteroides fragilis extract contains bacteroides fragilis capsular polysaccharide A.
In some embodiments, the bacteroides fragilis capsular polysaccharide a has a molecular weight of 5-75 KD.
In some embodiments, the bacteroides fragilis capsular polysaccharide A has a molecular weight of 15 KD-65 KD;
in some embodiments, the bacteroides fragilis capsular polysaccharide A has a molecular weight of 25 KD-55K.
In some embodiments, the bacteroides fragilis capsular polysaccharide a has a molecular weight of 35KD to 45KD.
In some of these embodiments, the bacteroides fragilis extract contains bacteroides fragilis capsular polysaccharide a in an amount of 60-75wt%.
In some embodiments, the bacteroides fragilis is bacteroides fragilis ZY-312 with collection number of CGMCC No.10685.
In some of these embodiments, the method of preparing the bacteroides fragilis extract comprises the steps of:
(1) Centrifuging and precipitating the bacteroides fragilis bacterial liquid after fermentation culture, collecting a first precipitate, adding water with the temperature of 65-72 ℃ into the first precipitate, adding a phenol solution after dissolving, keeping the temperature of 65-72 ℃, stirring for 25-35min, centrifuging, and collecting a first supernatant;
(2) Extracting the first supernatant collected in the step (1) by using ether to remove phenol, removing residual ether, and collecting an aqueous phase solution;
(3) Adding absolute ethyl alcohol into the aqueous phase solution collected in the step (2) until the final concentration of the ethyl alcohol is 75-85v/v%, precipitating with alcohol, centrifuging, collecting a second precipitate,
(4) And adding water into the second precipitate to prepare a suspension, adjusting the pH to 6.5-7.5, centrifuging, collecting a second supernatant, dialyzing to remove salt, and freeze-drying to obtain the bacteroides fragilis extract.
In some embodiments, the ratio of the water, the phenol solution and the first precipitate added to the first precipitate in step (1) is 3-5mL:3-5mL:1g; the mass concentration of the phenol solution is 70-80%.
In some embodiments, the alcohol precipitation in step (3) is alcohol precipitation at 0-8 ℃ for 8-16 hours.
In some embodiments, step (4) comprises: and (3) adding water into the second precipitate to prepare a suspension with the mass concentration of 8-12%, adding a glacial acetic acid aqueous solution with the mass concentration of 8-12%, heating to boil, stirring for reacting for 1.5-2.5 hours, adjusting the pH to 6.5-7.5, centrifuging, collecting a second supernatant, dialyzing to remove salt, and freeze-drying to obtain the bacteroides fragilis extract.
In some embodiments, the method for preparing a bacteroides fragilis extract further comprises the step of degrading: degrading the bacteroides fragilis extract obtained in the step (4) by using an ultrasonic method, wherein the ultrasonic condition is as follows: 180-210kHz,15-25 ℃.
In some of these embodiments, the dosage form of the medicament comprises a pill, tablet, granule, capsule, oral liquid, or tube feed formulation. The medicine comprises human medicine or animal medicine, and can be used for human or animal.
The medicament can comprise one or more of the following pharmaceutically acceptable auxiliary materials: diluents, excipients, binders, lubricants, suspending agents, coating agents, solubilizers, and the like. Examples of pharmaceutically acceptable excipients include: water, salt solutions, alcohols, silicones, waxes, petrolatum, vegetable oils, polyethylene glycols, propylene glycols, liposomes, saccharides, gelatin, lactose, amylose, magnesium stearate, talc, surfactants, silicic acid, viscous paraffin, perfume oil, mono-and di-fatty acid glycerides, petro (petrochemical) fatty acid esters, hydroxymethyl cellulose, polyvinylpyrrolidone, and the like.
The medicament may be administered by any one or more of the following: administration by inhalation in the form of a micro-pump or nasal spray or inhalation aerosol or the like, administration in the form of a suppository or pessary, topical administration in the form of a lotion, solution, cream, ointment or dusting powder, administration by use of a skin patch, oral administration in the form of a tablet containing an excipient such as starch or lactose or in the form of a capsule or an ovule alone or mixed with an excipient, or administration in the form of an elixir, solution or suspension containing flavouring or colouring agents, or parenteral injection, for example intracavernosal, intravenous, intramuscular or subcutaneous injection. For parenteral administration, the drug is preferably used in the form of a sterile aqueous solution which may contain other substances, for example enough salts or monosaccharides to make the solution isotonic with blood. For buccal or sublingual administration, the medicament may be administered in the form of tablets or lozenges formulated in conventional manner.
The bacteroides fragilis ZY-312 of the invention has been preserved in China general microbiological culture Collection center (CGMCC) at 4.2.2015, the preservation number is CGMCC No.10685, and the preservation address is No. 3 Hospital No.1 of Xilu, beijing, chaoyang district.
The inventor of the invention obtains a preparation method of the bacteroides fragilis extract (the main component of which is capsular polysaccharide A) through long-term experience accumulation and a large number of creative experimental researches, and further experimental researches show that the bacteroides fragilis extract has the function of preventing and treating irritable bowel syndrome, has good prevention and treatment effects on diarrhea-type irritable bowel syndrome and constipation-type irritable bowel syndrome, and has a far better prevention and treatment effect on the irritable bowel syndrome than that of the bacteroides fragilis. Further, the inventor obtains the bacteroides fragilis capsular polysaccharide A with the molecular weight of 5-70KD by degrading the bacteroides fragilis capsular polysaccharide A with the molecular weight of 70KD, and unexpectedly finds that the bacteroides fragilis capsular polysaccharide A with the molecular weight of 5-70KD has a better function of preventing and treating irritable bowel syndrome, and the effect of the bacteroides fragilis capsular polysaccharide A is far better than that of the bacteroides fragilis NCTC 9343. The bacteroides fragilis capsular polysaccharide A provided by the invention has a good prevention and treatment effect on irritable bowel syndrome and has no side effect on organisms, and can be used for performing preventive and therapeutic administration alone or together with other probiotics and/or probiotic materials. The Bacteroides fragilis capsular polysaccharide A provided by the invention has good edible and medicinal prospects, and provides a good product which is suitable for human body eating and is used for health care and irritable bowel syndrome prevention and treatment in clinic.
Drawings
FIG. 1 is a characteristic map of colony of Bacteroides fragilis ZY-312 of example 1;
FIG. 2 is a microscopic image of Bacteroides fragilis ZY-312 of example 1 after gram-staining.
FIG. 3 is a 1H spectrum of capsular polysaccharide A NMR spectroscopy analysis of example 1;
FIG. 4 is a 13C spectrum of capsular polysaccharide A NMR spectrometer analysis of example 1;
FIG. 5 is a COSY spectrum of capsular polysaccharide A NMR spectrometer analysis of example 1;
FIG. 6 is an HSQC spectrum of capsular polysaccharide A NMR spectrometer analysis of example 1;
FIG. 7 is a HMBC chromatogram of a nuclear magnetic resonance spectrometer analysis of capsular polysaccharide A from example 1;
FIG. 8 shows the chemical structure of Bacteroides fragilis capsular polysaccharide A prepared in example 1.
Detailed Description
The present invention will be described in further detail with reference to the following examples, which are provided for illustrative purposes only and do not limit the scope of the present invention.
The Bacteroides fragilis used in the following examples is Bacteroides fragilis ZY-312 (bacteriodes fragilis ZY-312), which was deposited in China general microbiological culture Collection center (CGMCC) at 4.2.2015, with the collection number of CGMCC No.10685, and the collection address of No. 3, xilu 1, north Cheng Yang district, beijing.
Example 1 preparation of Bacteroides fragilis extract
(1) Fermentation culture of bacteroides fragilis
The strain is streaked and inoculated on a blood plate, and anaerobic culture is carried out for 48h. Observing the morphological characteristics, staining characteristics, size, club shape, distribution and the like of colonies.
Colony characteristics: after culturing the bacteroides fragilis ZY-312 on a blood plate for 48 hours, the bacteroides fragilis ZY-312 presents a round convex shape, translucency, white color, smooth surface and no hemolysis, and the diameter of a colony is between 1 and 3mm, see figure 1.
Microscopic morphology: bacteroides fragilis ZY-312 was subjected to gram-staining microscopic examination, and was gram-negative bacteria, which were typically rod-shaped, had blunt and densely stained at both ends, and had no staining part in the middle of the bacteria, such as vacuole, as shown in FIG. 2.
Selecting single colony, inoculating into tryptone broth, fermenting and culturing for 8 hours (at 37 ℃), centrifuging the obtained bacterial liquid for precipitation, rotating at 3000r/min, centrifuging for 15min, removing supernatant, and collecting precipitate.
(2) Preparation of Bacteroides fragilis extract
1) Taking 200g of bacteroides fragilis bacterial mud (the precipitate obtained in the step (1)) and adding 750mL of ultrapure water at 68 ℃, after dissolving, adding 750mL of 75% phenol solution by volume fraction, uniformly mixing, keeping the temperature at 68 ℃, stirring and extracting for 30min, centrifuging for 20min at 15000g, and taking supernatant.
2) The supernatant was extracted with an equal volume of ether (1.5L) to remove phenol, the supernatant collected and extracted repeatedly until no phenol remained. The ether was removed by heating in a water bath and the aqueous phase was collected.
3) Centrifuging the water phase at 15000g for 20min, measuring volume, adding anhydrous ethanol until the final concentration of ethanol is 80% (volume fraction), precipitating with ethanol at 4 deg.C overnight (12 hr), centrifuging at 15000g for 20min, and collecting precipitate.
4) Weighing the mass of the precipitate in the step 3), adding a certain volume of deionized water to prepare the precipitate into a suspension with the mass concentration of 10%, stirring and mixing uniformly, adding a glacial acetic acid aqueous solution with the mass concentration of 10%, heating to boil, continuously stirring and reacting for 2h, adjusting the pH value to 7.0, centrifuging for 20min at 15000g, and collecting the supernatant. Dialyzing the obtained supernatant to remove salt (10 KD dialysis bag), and freeze-drying to obtain Bacteroides fragilis extract.
5) Weighing 30mg of Bacteroides fragilis extract of step 4) in 0.5mL D 2 O, 1. Mu.l of acetone (1H, 2.22, 13C, 30.89) was added for calibration. Analyzing the spectra of 1H, 13C, COSY, HSQC and HMBC (figure 3) by using a 500MHz Bruker nuclear magnetic resonance spectrometer, and confirming that the bacteroides fragilis extract collected in the step 4) is capsular polysaccharide A and the purity is about 70%. GPC (gel permeation chromatography) analysis shows that the repeating unit of the capsular polysaccharide A has a molecular weight of 781, a repeating number n of 89, a molecular weight of about 70KD and a molecular formula of- [ C ] 31 N 3 O 20 H 47 ] 91 The chemical structure is shown in FIG. 4.
(3) Preparation of capsular polysaccharide A of different molecular weights
This example is carried out by degrading capsular polysaccharide A prepared in (2) by methods including, but not limited to, chemical degradation, physical degradation and biodegradation. In the embodiment, an ultrasonic method is adopted, wherein the capsular polysaccharide A is treated for 3 hours, 2 hours, 0.5 hour and 0 hour at 195kHz and 20 ℃, and capsular polysaccharide A with molecular weights of 2KD, 5KD, 40KD and 70KD is collected respectively. Capsular polysaccharide A with molecular weight of 110KD is extracted from Bacteroides fragilis NCTC 9343 (purchased from ATCC in USA) by the method of (2).
Example 2 Effect of Bacteroides fragilis capsular polysaccharide A on diarrhea-causing IBS in folium sennae
1. Design of experiments
In order to verify the effect of the bacteroides fragilis extract (capsular polysaccharide a as a main ingredient) provided in example 1 on the prevention/treatment of irritable bowel syndrome, 60C 57BL/6 mice were selected for the experiment. Each of 60C 57BL/6 mice was hermaphrodite, and each experimental mouse was assigned a unique number. Before animals are grouped, the squirrel cage labels should be labeled with item number, species/strain, gender, cage number, and animal number. The mice were randomly grouped according to the initial body weight of C57BL/6 mice using BioBook software, and were divided into 6 groups, namely a normal Group (Group 1), a model Group (Group 2), a loperamide capsule Group (2.4 mg/kg) (Group 3), a Bacteroides fragilis capsular polysaccharide A low (Group 4), medium (Group 5), and high (Group 6) dose groups, with 10 mice per Group C57 BL/6. This example illustrates capsular polysaccharide A, which has a molecular weight of 5KD.
Mice in each group were gavaged with the corresponding drug, and normal and model groups were given an equal amount of physiological saline 1 time a day for 5 consecutive days. All C57BL/6 mice were fasted for 24h, and 1h after the last administration, 1g/mL senna leaf decoction was administered to each Group of C57BL/6 mice, except for the normal Group (Group 1) which was administered with the same amount of physiological saline. The C57BL/6 mice are raised in cages, 1 mouse is fed in each cage, filter paper is padded under the cages to count wet manure, the diarrhea degree is represented by the amount of the wet manure, and the filter paper is changed for 1 time every 1 h. And observing and counting the total number of stools, the total number of loose stools and the number of the loose stool series of the mice within 6 h.
2. Criterion of evaluation
The number of rare earths was determined according to the Zhou's method (Zhou gan, hu Zhi Hua, wang Yao, et al. Preparation of a model of diarrhea in mice and use of the index of diarrhea [ J ]. Chinese herbal medicine, 1994, 250 (4): 195-196.). The diameter of the stain is 1 grade when the diameter is less than l cm, 2 grade when the diameter is 1-1.9 cm, 3 grade when the diameter is 2-3 cm, and 4 grade when the diameter is more than 3 cm. The stool dilution rate (%) = total number of stool/total number of stools × 100% per animal, and the diarrhea Index (ID) = stool dilution rate × stool dilution number.
3. Results and analysis
All data are expressed in x +/-s, statistical analysis is carried out by using SPSS 17.0 software, single-factor analysis of variance is adopted for comparison among groups, and P is less than 0.05, so that statistical significance is achieved. Specific results are shown in table 1.
TABLE 1 Effect of Bacteroides fragilis capsular polysaccharide A on diarrhea-predominant IBS in mice caused by folium sennae: (n=10)
Note: P < 0.01 compared to normal group; compared with the model group, P is less than 0.01.
As can be seen from the results in Table 1, the total number of loose stools, the rate of loose stools and the diarrhea index comparison between the model Group (Group 2) and the normal Group (Group 1) are all significantly different (P < 0.01), which indicates that the IBS model of the mice diarrhea caused by senna leaves is successfully modeled. Compared with the model group, the loperamide group and the bacteroides fragilis capsular polysaccharide A low, medium and high dose groups provided in example 1 can obviously inhibit the degree of diarrhea (P is less than 0.01) of mice, and the bacteroides fragilis capsular polysaccharide A provided by the invention has obvious inhibition effect on diarrhea-type IBS caused by folium sennae.
Example 3 Effect of Bacteroides fragilis capsular polysaccharide A on Castor oil-induced diarrheal IBS
1. Design of experiments
In order to verify the effect of the bacteroides fragilis extract (capsular polysaccharide a as a main ingredient) provided in example 1 on the prevention/treatment of irritable bowel syndrome, 60C 57BL/6 mice were selected for the experiment. The 60C 57BL/6 mice are hermaphroditic, and each experimental mouse is assigned a unique number. Before animals are grouped, the squirrel cage labels should be labeled with item number, species/strain, gender, cage number, and animal number. The mice were randomly grouped according to the initial body weight of C57BL/6 mice using BioBook software, and were divided into 6 groups, i.e., normal Group (Group 1), model Group (Group 2), loperamide capsule Group (2.4 mg/kg) (Group 3), bacteroides fragilis capsular polysaccharide A low (Group 4), medium (Group 5), and high (Group 6) dose groups, each of which contained 10 mice per Group C57 BL/6. This example illustrates capsular polysaccharide A, which has a molecular weight of 40 kD.
Mice in each group were gavaged with the corresponding drug, and normal and model groups were given an equal amount of physiological saline 1 time a day for 5 consecutive days. All C57BL/6 mice were fasted for 24h, and 1h after the last administration, except for the normal Group (Group 1), the C57BL/6 mice in each Group were administered castor oil 1 time (20 mL/kg). The C57BL/6 mice are raised in cages, 1 mouse is fed in each cage, filter paper is padded under the cages to count wet manure, the diarrhea degree is represented by the amount of the wet manure, and the filter paper is changed for 1 time every 1 h. And observing and counting the total number of shit, the total number of shit and the number of shit stages of the mice within 6 h.
2. Criterion of evaluation
Same as the criteria of example 2.
3. Results and analysis
All data are expressed inIt shows that SPSS 17.0 software is used for statistical analysis, single-factor analysis of variance is adopted for comparison among groups, and P is less than 0.05, which is of statistical significance. Specific results are shown in table 2.
TABLE 2 Effect of Bacteroides fragilis capsular polysaccharide A on Castor oil-induced diarrhea-type IBS in mice: (n=10)
Note P < 0.01 compared to normal group; compared with the model group, P is less than 0.01.
As can be seen from the results in Table 2, the total number of loose stools, the rate of loose stools and the diarrhea index comparison of the model Group (Group 2) and the normal Group (Group 1) all have significant differences (P < 0.01), which indicates that the IBS model of the mice diarrhea caused by castor oil is successfully modeled. Compared with the model group, the loperamide group can obviously inhibit the total number of loose stools of the mice with castor oil-induced diarrhea for 6 hours, the rate of the loose stools and the diarrhea index (P < 0.05). The low dose (0.125 g/kg) of Bacteroides fragilis capsular polysaccharide A has certain inhibition effect on total number of loose stools, loose stool rate and diarrhea index after 6 hours compared with the model group, but has no significant difference (P is more than 0.05); but in medium and high dose, the bacteroides fragilis capsular polysaccharide A can remarkably inhibit the total number of loose stools of 6h, the loose stool rate and the diarrhea index (P < 0.05) of mice with castor oil diarrhea.
Example 4 Effect of Bacteroides fragilis capsular polysaccharide A on neostigmine-induced hyperkinetic movement of mouse Small intestine
1. Design of experiments
In order to verify the effect of the bacteroides fragilis extract (capsular polysaccharide a as the main ingredient) provided in example 1 on the prevention/treatment of irritable bowel syndrome, 60C 57BL/6 mice were selected for the experiment in this example. The 60C 57BL/6 mice are hermaphroditic, and each experimental mouse is assigned a unique number. Before animals are grouped, the squirrel cage labels should be labeled with item number, species/strain, gender, cage number, and animal number. The mice were randomly grouped according to the initial body weight of C57BL/6 mice using BioBook software into 6 groups, namely a normal Group (Group 1), a model Group (Group 2), a pinaverium bromide (0.1 g/kg) Group (Group 3), a bacteroides fragilis capsular polysaccharide a low (Group 4), medium (Group 5), high (Group 6) dose Group, and 10 mice per Group, C57 BL/6. This example illustrates capsular polysaccharide A, which has a molecular weight of 70 kD.
Mice in each group were gavaged with the corresponding drug, and normal and model groups were given an equal amount of physiological saline 1 time a day for 5 consecutive days. 1h after the last administration, except for the normal group, neostigmine was subcutaneously injected in 5 groups at 0.15mg/kg, causing the small intestine to be too excitable, and the normal group was subcutaneously injected with an equal amount of physiological saline. After 15min, the groups were gazed with a suspension containing 5% charcoal dust, and after 20min, the cervical vertebrae were taken off and sacrificed, the small intestine was separated by opening the abdomen, the total length of the small intestine and the length of the small intestine through which the charcoal dust pushed were measured, and the percentage of the charcoal dust pushed was calculated. Percent carbon-pushed = (length of small intestine pushed by carbon/total length of small intestine) × 100%.
2. Results and analysis
All data are as followsIt shows that SPSS 17.0 software is used for statistical analysis, single-factor analysis of variance is adopted for comparison among groups, and P is less than 0.05, which is of statistical significance. Specific results are shown in table 3.
TABLE 3 Effect of Bacteroides fragilis capsular polysaccharide A on neostigmine-induced hypermotility of mouse small intestine: (n=10)
Note P < 0.01 compared to normal group; compared with the model group, P is less than 0.01.
As can be seen from Table 3, neostigmine can cause the mouse small intestine hyperkinesia, compared with the model group, the pinaverium bromide group can obviously reduce the carbon-end propulsion rate and inhibit the mouse small intestine hyperkinesia caused by neostigmine (P is less than 0.01); the Bacteroides fragilis capsular polysaccharide A with medium and high dose can also remarkably reduce carbon dust propulsion rate (P < 0.05), and inhibit mouse intestinal hypermotility caused by neostigmine.
Example 5 Effect of Bacteroides fragilis capsular polysaccharide A on Constipation-type IBS
1. Design of experiments
In order to verify the effect of the bacteroides fragilis extract (capsular polysaccharide a as a main ingredient) provided in example 1 on prevention/treatment of constipation-predominant irritable bowel syndrome, 60 SD rats were selected for experiments in this example. The 60 SD rats are half male and half female, and each experimental rat is assigned a unique number. Before animals are grouped, the squirrel cage labels should be labeled with item number, species/strain, gender, cage number, and animal number. Groups 6, namely a normal Group (Group 1), a model Group (Group 2), a tegaserod maleate (solution preparation: 1.2mg tegaserod maleate added with 10ml sterile physiological saline) Group (Group 3), a bacteroides fragilis capsular polysaccharide A low (Group 4), medium (Group 5) and high (Group 6) dose Group, 10 SD rats in each Group were randomly divided according to the initial weight of the SD rats by using BioBook software. This example illustrates capsular polysaccharide A, which has a molecular weight of 70 kD.
SD rat constipation type IBS model was constructed according to the method of (Pen L H, yang Y S, sun G, et al. A new model of coherence-preventive bilateral synthesis in rats [ J ]. World core Journal of dictionary, 2004,12 (1): 112-116.). Except for the normal Group (Group 1), the rest groups are subjected to intragastric administration once a day by using ice physiological saline (0-4 ℃), 2ml for each time and 14 days continuously, and a constipation type rat IBS model is established. During the molding period, rats in each group can drink water freely.
After 14 days, each test group is administered with the corresponding drug by intragastric administration, wherein the normal group and the model group are administered with 10ml of normal-temperature sterile normal saline respectively; the positive control Group (Group 3) is filled with tegaserod maleate; groups 4-6 administered low, medium and high doses of bacteroides fragilis capsular polysaccharide a, respectively, specific experiments and dosing regimens are shown in table 4:
table 4 experimental groups and dosing regimens
2. Results and analysis
The number of fecal particles in each group of rats was collected at 24 hours on day 1, 14 and 28, respectively, and one contamination blot was used as one if any diarrhea was observed (see Table 5 for details). The collected feces were weighed, dried and the moisture content of the feces calculated (see table 6 for details).
| Day | 1 | Day 14 | |
Group1 | 49.13±6.34 | 48.53±6.48 | 48.10±5.43 ◆ | |
Group2 | 47.37±5.27 | 33.07±5.32 *▲ | 32.44±6.23 | |
Group3 | 48.22±7.34 | 32.46±5.72 *▲ | 47.43±6.35 ★◆ | |
Group4 | 47.78±5.39 | 32.73±4.39 *▲ | 40.04±7.94 ★◆ | |
Group5 | 48.05±7.91 | 34.07±5.83 *▲ | 43.90±8.06 ★◆ | |
Group6 | 47.88±6.54 | 33.77±5.60 *▲ | 49.03±5.71 ★◆ |
Note that P < 0.01 for each group of experiments compared to the first day; on the 14 th day, the ratio of Group 2-6 to the normal control Group is that a-P is less than 0.01; the ratio of: (28) to (14) in each experiment,: (0.01) major component; on day 28, each experimental group was compared to the model group, P < 0.01.
Note that P < 0.05 for each group of experiments compared to the first day; on the 14 th day, the ratio of Group 2-6 to the normal control Group is that a-P is less than 0.05; the ratio of day 28 to day 14 for each set of experiments,: < 0.05; on day 28, each experimental group was compared to the model group, P < 0.05.
As can be seen from Table 5, in groups 1-6, the number of fecal particles was not greatly different in the first 24 hours of day for each Group of rats; the number of fecal particles in the rats of the other groups on day 14 is obviously reduced compared with that of the rats on the first day except the normal group, and the difference has statistical significance (P is less than 0.05); on day 14, group 2-6 showed a significant decrease in stool particle count compared to the normal Group (Group 1), with the differences having statistical significance (P < 0.05), indicating that this example successfully constructed a constipation-type IBS rat model. On day 28, the number of fecal particles from groups 2-6 was significantly increased compared to day 14, with the difference being statistically significant (P < 0.05). The low, medium and high doses of bacteroides fragilis capsular polysaccharide A provided by the invention can effectively increase the number of fecal particles of constipation-type IBS rats.
As can be seen from Table 6, in groups 1-6, the water content of the feces is not greatly different in the first 24 hours of the day of the rats in each Group; except for the normal Group, the water content of the feces of the rats in the other groups on the 14 th day is obviously reduced compared with that of the rats on the first day, the difference has statistical significance (P is less than 0.05), and on the 14 th day, the water content of the feces of the groups 2-6 is obviously reduced compared with that of the rats in the normal Group (Group 1), and the difference has statistical significance (P is less than 0.05). On day 28, stool moisture content of groups 2-6 was significantly increased compared to day 14, with the difference being statistically significant (P < 0.05). The low, medium and high doses of bacteroides fragilis capsular polysaccharide A provided by the invention can effectively increase the water content of feces of constipation IBS rats.
From the results, the bacteroides fragilis capsular polysaccharide A provided by the invention has a good treatment effect on constipation type IBS.
Example 6 therapeutic Effect of Bacteroides fragilis and Bacteroides fragilis capsular polysaccharide A
1. Design of experiments
In order to compare the effect of the bacteroides fragilis extract (mainly comprising capsular polysaccharide a) provided in example 1 and bacteroides fragilis itself on the prevention/treatment of irritable bowel syndrome, 60C 57BL/6 mice were selected for the experiment in this example. The 60C 57BL/6 mice are hermaphroditic, and each experimental mouse is assigned a unique number. Before animals are grouped, the squirrel cage labels should be labeled with item number, species/strain, gender, cage number, and animal number. The mice were randomly assigned using BioBook software based on initial body weights of C57BL/6 mice into 6 groups, i.e., senna diarrhea-causing IBS model Group (Group 1), castor oil diarrhea-causing IBS model Group (Group 2), high-dose bacteroides fragilis capsular polysaccharide a versus treatment Group (Group 3) for senna diarrhea-causing IBS, high-dose bacteroides fragilis itself (10) 10 CFU/ml) treatment Group for diarrhea-causing IBS with senna (Group 4), high dose Bacteroides fragilis capsular polysaccharide A for diarrhea-causing IBS with castor oil (Group 5), and high dose Bacteroides fragilis itself (10) 10 CFU/ml) treatment Group for castor oil diarrhea IBS (Group 6), 10 mice per Group C57 BL/6. The molecular weight of capsular polysaccharide A in the embodiment is 40KD, and the concentration is 0.5mg/mL; the concentration of Bacteroides fragilis is 10 10 CFU/ml。
The mice in each Group were gavaged with the corresponding drug, and the model groups (Group 1, group 2) were given an equal amount of physiological saline 1 time a day for 5 consecutive days. All C57BL/6 mice were fasted for 24h, 1g/mL senna leaf water decoction was administered to 1h, group1, group3 and Group 4C 57BL/6 mice after the last dose, and castor oil was administered to Group2, group5 and Group 6C 57BL/6 mice 1 time (20 mL/kg). The C57BL/6 mice are raised in cages with 1 mouse per cage, filter paper is padded under the cages for counting wet manure, the degree of diarrhea is represented by the amount of the wet manure, and the filter paper is replaced for 1 time every 1 h. And observing and counting the total number of shit, the total number of shit and the number of shit stages of the mice within 6 h.
2. Criterion of evaluation
Same as the criteria of example 2.
3. Results and analysis
All data are as followsIt shows that SPSS 17.0 software is used for statistical analysis, single-factor analysis of variance is adopted for comparison among groups, and P is less than 0.05, which is of statistical significance. Specific results are shown in table 6.
Note: group3 and Group4 are respectively compared with Group1, P is less than 0.05; group5, group6 and Group2, ▲ p is less than 0.05; group3 compared to Group4, group5 compared to Group6, ★ P<0.05。
as can be seen from the results in Table 6, the total number of loose stools, the rate of loose stools and the diarrhea index comparison of the Group3 and the Group4 with the model Group (Group 1) are all significantly different (P < 0.05), which indicates that the Bacteroides fragilis per se and the Bacteroides fragilis capsular polysaccharide A have treatment and prevention effects on IBS caused by senna leaves in mice. Compared with the total number of loose stools, the loose stool rate and the diarrhea index of the Group5 and the Group6 (Group 2), the significant differences (P is less than 0.01) are shown, which indicates that the bacteroides fragilis and the bacteroides fragilis capsular polysaccharide A have treatment and prevention effects on IBS caused by castor oil. Compared with Group4, the total number of Group3 loose stools, the loose stool rate and the diarrhea index are small and have significant differences (P is less than 0.05), which indicates that the Bacteroides fragilis capsular polysaccharide A has better treatment and prevention effects on IBS (IBS) caused by diarrhea of mice due to senna leaves than the Bacteroides fragilis per se; compared with Group6, the total number of loose stools, the loose stool rate and the diarrhea index of Group5 are small and have significant differences (P is less than 0.05), which indicates that the Bacteroides fragilis capsular polysaccharide A has better IBS treatment and prevention effects on castor oil-induced diarrhea of mice than Bacteroides fragilis per se.
Experiments also prove that the bacteroides fragilis capsular polysaccharide A has better treatment and prevention effects on mouse small intestine hyperkinetic function and constipation type IBS caused by neostigmine than the bacteroides fragilis.
Example 7 therapeutic Effect of Bacteroides fragilis capsular polysaccharide A of different molecular weights on IBS
In this example, the bacteroides fragilis extracts containing bacteroides fragilis capsular polysaccharide A with 2KD, 5KD, 40KD and 70KD prepared in the invention example 1 were used for preventing/treating constipation type IBS rat model, and the therapeutic effect of bacteroides fragilis capsular polysaccharide A with different molecular weights on constipation type IBS was examined. This example illustrates high doses of 2kD, 5kD, 40kD and 70kD Bacteroides fragilis capsular polysaccharide A.
1. Design of experiments
Referring to the experimental grouping method of example 5, mice were divided into a normal control group, a model group, a 2KD group, a 5KD group, a 40KD group, a 70KD group, and a 110KD group.
The SD rat constipation-type IBS model was constructed according to the method of (Peng L H, yang Y S, sun G, et al. A new model of coherent-predominable bowelsynthesis in rates [ J ]. World Chinese Journal of dictionary, 2004,12 (1): 112-116.). Except for the normal Group (Group 1), the other groups are perfused with ice physiological saline (0-4 ℃) once a day, 2ml each time, and continuously for 14 days, so as to establish a constipation type rat IBS model. During the molding period, rats in each group can drink water freely.
After 14 days, each test group was administered the corresponding drug by gavage, wherein the normal group and the model group were administered 10ml of normal temperature sterile physiological saline, respectively; groups 3-7 were administered capsular polysaccharides a with molecular weights of 2KD (Group 3), 5KD (Group 4), 40KD (Group 5), 70KD (Group 6) and 110KD (Group 7), respectively, and the specific experiments and dosing schedule are shown in table 7:
table 7 experimental groups and dosing regimens
2. Results and analysis
The number of fecal particles in each group of rats was collected at 24 hours on day 1, 14 and 28, respectively, and one contamination blot was used as one if any diarrhea was observed (see Table 8 for details). The collected feces were weighed, dried, and the moisture content of the feces was calculated (see table 9 for details).
Note that P < 0.01 for each group of experiments compared to the first day; on the 14 th day, the ratio of Group 2-7 to the normal control Group is that a-P is less than 0.01; the ratio of: (28) to (14) in each experiment,: (0.01) major component; on day 28, each experimental group was compared to the model group,. Diamond-solid.P < 0.01.
| Day | 1 | Day 14 | |
Group1 | 0.50±0.03 | 0.48±0.05 | 0.49±0.08 ◆ | |
Group2 | 0.49±0.06 | 0.37±0.04 *▲ | 0.34±0.05 | |
Group3 | 0.48±0.08 | 0.38±0.07 *▲ | 0.41±0.07 ★◆ | |
Group4 | 0.48±0.07 | 0.36±0.06 *▲ | 0.54±0.08 ★◆ | |
Group5 | 0.49±0.09 | 0.37±0.10 *▲ | 0.53±0.06 ★◆ | |
Group6 | 0.48±0.08 | 0.39±0.08 *▲ | 0.55±0.09 ★◆ | |
Group7 | 0.47±0.08 | 0.38±0.05 *▲ | 0.45±0.08 ★◆ |
Note that P < 0.05 for each group of experiments compared to the first day; on the 14 th day, comparing Group 2-6 with normal control Group, P is less than 0.05; the ratio of the first day 28 to the second day 14 in each experiment is: < 0.05; on day 28, each experimental group was compared to the model group, P < 0.05.
As can be seen from Table 8, in groups 1-7, the number of fecal particles was not greatly different in the first 24 hours of day for each Group of rats; except for the normal Group (Group 1), the number of the fecal particles of the rats in other groups on the 14 th day is obviously reduced compared with that of the rats on the first day, the difference is extremely obvious (P is less than 0.01), and the statistical significance is achieved; on day 14, group 2-7 were significantly reduced compared to the number of fecal particles in the normal Group (Group 1), with significant differences (P < 0.01), and statistical significance. This example illustrates the successful construction of a constipation-predominant IBS rat model. On the 28 th day, except for Group3, the numbers of the fecal particles of groups 4-7 are obviously increased compared with those on the 14 th day, the difference is extremely obvious (P is less than 0.01), and the statistical significance is achieved. The bacteroides fragilis capsular polysaccharide A with different molecular weights provided by the invention can effectively increase the number of fecal particles of constipation type IBS rats. Meanwhile, the fecal granule number of the groups 3-7 in 28 days is compared to find that the fecal granule number of the groups 4, 5 and 6 is obviously more than that of the groups 2 and 7, the difference is extremely obvious (P is less than 0.05), and the statistical significance is achieved.
As can be seen from Table 9, in groups 1-7, the moisture content of the feces is not much different in the first 24 hours of the day of each Group of rats; except for the normal Group, the water content of the feces of the rats in the other groups on the 14 th day is obviously reduced compared with that of the rats on the first day, the difference has statistical significance (P is less than 0.05), and on the 14 th day, the water content of the feces of the groups 2-7 is obviously reduced compared with that of the rats in the normal Group (Group 1), and the difference has statistical significance (P is less than 0.05). On day 28, with the exception of Group3, the fecal moisture content of groups 4-7 was significantly increased compared to that on day 14, with the difference being statistically significant (P < 0.05). Meanwhile, the water content of the feces among the groups 3-7 in 28 days is compared, and the feces water content of the groups 4, 5 and 6 is obviously more than that of the groups 3 and 7, the difference is extremely obvious (P is less than 0.05), and the statistical significance is achieved.
The results show that the bacteroides fragilis capsular polysaccharide A with the molecular weight of 5 KD-70 KD provided by the invention can effectively increase the stool particle number and the stool water content of constipation type IBS rats, and has a good treatment effect on constipation type IBS. Meanwhile, the degradation of the Bacteroides fragilis capsular polysaccharide A is also demonstrated, so that the molecular weight and viscosity of the capsular polysaccharide A are reduced, and the treatment effect on constipation-predominant irritable bowel syndrome can be enhanced.
Meanwhile, experiments prove that the bacteroides fragilis capsular polysaccharide A with the molecular weight of 5 KD-70 KD has better effect on reducing diarrhea index of diarrhea-type IBS than the bacteroides fragilis capsular polysaccharide A with the molecular weight of 2KD or 110 KD.
Example 8 therapeutic Effect of different Bacteroides fragilis strains capsular polysaccharide A on Constipation-type IBS
This example used the sonication method described in example 1 to degrade capsular polysaccharide A with molecular weight of 110kD (sonication conditions: 195kHz,25 ℃,0.5 hour), and collected capsular polysaccharide A with molecular weight of 70kD, which was designated NCTC 9343-70kD group, and compared with capsular polysaccharide A with molecular weight of 70kD extracted from ZY-312 (designated ZY-312-70kD group), to evaluate the therapeutic effect on constipation-type IBS. In this example, referring to the method described in example 7, the change of the number of fecal particles and the change of the water content of rat feces were detected, and the specific results are as follows:
Note that P < 0.01 for each group of experiments compared to the first day; the ratio of day 28 to day 14 for each set of experiments, ≧ 0.01.
Note that P < 0.01 for each group of experiments compared to the first day; the ratio of day 28 to day 14 for each set of experiments, ≧ 0.01.
From the above results, it can be seen that the capsular polysaccharide A with molecular weight of 110KD extracted from the NCTC 9343 strain is degraded, and the therapeutic effect similar to that of the capsular polysaccharide A extracted from ZY-312 strain on constipation IBS can be realized.
The results of the above examples show that the bacteroides fragilis capsular polysaccharide A provided by the invention has good prevention and treatment effects on diarrhea type and constipation type IBS, and has a bidirectional regulation effect.
All possible combinations of the technical features of the above embodiments may not be described for the sake of brevity, but should be considered as within the scope of the present disclosure as long as there is no contradiction between the combinations of the technical features.
The above-mentioned embodiments only express several embodiments of the present invention, and the description thereof is specific and detailed, but not to be understood as limiting the scope of the invention. It should be noted that various changes and modifications can be made by those skilled in the art without departing from the spirit of the invention, and these changes and modifications are all within the scope of the invention. Therefore, the protection scope of the present patent should be subject to the appended claims.
Claims (1)
1. The application of Bacteroides fragilis (Bacteroides fragilis) ZY-312 in preparing a medicine for preventing and/or treating constipation-predominant irritable bowel syndrome is characterized in that the preservation number of the Bacteroides fragilis is CGMCC No.10685.
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202110977532.0A CN113730443B (en) | 2017-09-11 | 2017-09-11 | Application of bacteroides fragilis and extract thereof in preparation of medicine for preventing and treating irritable bowel syndrome |
Applications Claiming Priority (2)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN201710812676.4A CN109481473B (en) | 2017-09-11 | 2017-09-11 | Application of bacteroides fragilis extract in preparation of medicine for preventing and treating irritable bowel syndrome |
CN202110977532.0A CN113730443B (en) | 2017-09-11 | 2017-09-11 | Application of bacteroides fragilis and extract thereof in preparation of medicine for preventing and treating irritable bowel syndrome |
Related Parent Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN201710812676.4A Division CN109481473B (en) | 2017-09-11 | 2017-09-11 | Application of bacteroides fragilis extract in preparation of medicine for preventing and treating irritable bowel syndrome |
Publications (2)
Publication Number | Publication Date |
---|---|
CN113730443A CN113730443A (en) | 2021-12-03 |
CN113730443B true CN113730443B (en) | 2022-10-04 |
Family
ID=65633485
Family Applications (3)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202110977532.0A Active CN113730443B (en) | 2017-09-11 | 2017-09-11 | Application of bacteroides fragilis and extract thereof in preparation of medicine for preventing and treating irritable bowel syndrome |
CN202210968927.9A Pending CN115252652A (en) | 2017-09-11 | 2017-09-11 | Application of bacteroides fragilis in preparation of medicine for preventing and treating irritable bowel syndrome |
CN201710812676.4A Active CN109481473B (en) | 2017-09-11 | 2017-09-11 | Application of bacteroides fragilis extract in preparation of medicine for preventing and treating irritable bowel syndrome |
Family Applications After (2)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202210968927.9A Pending CN115252652A (en) | 2017-09-11 | 2017-09-11 | Application of bacteroides fragilis in preparation of medicine for preventing and treating irritable bowel syndrome |
CN201710812676.4A Active CN109481473B (en) | 2017-09-11 | 2017-09-11 | Application of bacteroides fragilis extract in preparation of medicine for preventing and treating irritable bowel syndrome |
Country Status (2)
Country | Link |
---|---|
CN (3) | CN113730443B (en) |
WO (1) | WO2019047439A1 (en) |
Families Citing this family (12)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN113651896B (en) * | 2021-07-09 | 2022-07-12 | 中山大学 | Bacteroides dychii extracellular polysaccharide and extraction method and application thereof |
CN115957243A (en) * | 2021-10-12 | 2023-04-14 | 广州知易生物科技有限公司 | Application of bacteroides fragilis in preventing and treating cancer-related diarrhea |
CN114470003B (en) * | 2022-01-12 | 2023-07-25 | 广州知易生物科技有限公司 | Application of bacteroides fragilis or zwitterionic capsular polysaccharide thereof in preparing medicines for preventing and treating digestive system tumors |
CN114452382B (en) * | 2022-01-12 | 2023-07-18 | 广州知易生物科技有限公司 | Application of bacteroides fragilis capsular polysaccharide A and PD-1 and PD-L1 antibody in combined treatment of respiratory system tumor |
CN114344325B (en) * | 2022-01-12 | 2023-07-14 | 广州知易生物科技有限公司 | Application of bacteroides fragilis and zwitterionic capsular polysaccharide thereof in preparation of medicine for preventing and treating genitourinary system tumors |
CN114404598B (en) * | 2022-01-12 | 2023-07-18 | 广州知易生物科技有限公司 | Application of bacteroides fragilis capsular polysaccharide A combined with PD-1 inhibitor in preparation of medicines for treating skin tumor |
CN114558036A (en) * | 2022-01-25 | 2022-05-31 | 广州知易生物科技有限公司 | Application of bacteroides fragilis in improvement and treatment of diarrhea |
CN114699423B (en) * | 2022-02-16 | 2023-06-23 | 广州知易生物科技有限公司 | Application of capsular polysaccharide extract of bacteroides fragilis in preparation of medicines for preventing and treating schizophrenia |
CN114699425B (en) * | 2022-02-16 | 2023-08-04 | 广州知易生物科技有限公司 | Novel application of capsular polysaccharide A extract of bacteroides fragilis |
CN114699422B (en) * | 2022-02-16 | 2023-07-18 | 广州知易生物科技有限公司 | Application of capsular polysaccharide extract of bacteroides fragilis in preparation of medicines for preventing and treating Alzheimer disease |
CN114699424B (en) * | 2022-02-16 | 2023-07-18 | 广州知易生物科技有限公司 | New application of bacteroides fragilis zwitterionic capsular polysaccharide and/or modified zwitterionic capsular polysaccharide |
CN116004486B (en) * | 2023-03-24 | 2023-06-30 | 广东省科学院微生物研究所(广东省微生物分析检测中心) | Bacteroides fragilis BFS17 for relieving irritable bowel syndrome and intestinal tract hypersensitivity and application thereof |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013052099A2 (en) * | 2011-10-03 | 2013-04-11 | Mazmanian Sarkis K | Generation of psa-only producing mutant strain |
CN103082294A (en) * | 2013-01-21 | 2013-05-08 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for treating diarrhea |
CN103156888A (en) * | 2013-03-18 | 2013-06-19 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for treating inflammatory bowel diseases |
CN105434476A (en) * | 2015-10-29 | 2016-03-30 | 广州知易生物科技有限公司 | Application of bacteroides fragilis to prevention and/or treatment of inflammatory bowel diseases (IBDs) |
Family Cites Families (2)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
US9539281B2 (en) * | 2011-07-12 | 2017-01-10 | The Brigham And Women's Hospital, Inc. | Lipid-containing PSA compositions, methods of isolation and methods of use thereof |
MX2015011700A (en) * | 2013-03-05 | 2016-07-20 | Univ Groningen | Use of faecali bacterium prausnitzii htf-f (dsm 26943) to suppress inflammation. |
-
2017
- 2017-09-11 CN CN202110977532.0A patent/CN113730443B/en active Active
- 2017-09-11 CN CN202210968927.9A patent/CN115252652A/en active Pending
- 2017-09-11 CN CN201710812676.4A patent/CN109481473B/en active Active
- 2017-12-29 WO PCT/CN2017/120112 patent/WO2019047439A1/en active Application Filing
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
WO2013052099A2 (en) * | 2011-10-03 | 2013-04-11 | Mazmanian Sarkis K | Generation of psa-only producing mutant strain |
CN103082294A (en) * | 2013-01-21 | 2013-05-08 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for treating diarrhea |
CN103156888A (en) * | 2013-03-18 | 2013-06-19 | 广州知光生物科技有限公司 | Application of bacteroides fragilis in preparation of composition for treating inflammatory bowel diseases |
CN105434476A (en) * | 2015-10-29 | 2016-03-30 | 广州知易生物科技有限公司 | Application of bacteroides fragilis to prevention and/or treatment of inflammatory bowel diseases (IBDs) |
Non-Patent Citations (1)
Title |
---|
肠易激综合征与炎症性肠病;桑玉尔等;《国际消化病杂志》;20061225;第26卷(第6期);第384-386页 * |
Also Published As
Publication number | Publication date |
---|---|
CN113730443A (en) | 2021-12-03 |
CN109481473A (en) | 2019-03-19 |
WO2019047439A1 (en) | 2019-03-14 |
CN115252652A (en) | 2022-11-01 |
CN109481473B (en) | 2023-03-21 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN113730443B (en) | Application of bacteroides fragilis and extract thereof in preparation of medicine for preventing and treating irritable bowel syndrome | |
CN109789173B (en) | Nanovesicles derived from bacillus bacteria and uses thereof | |
US7785581B2 (en) | Composition and method for reducing feces toxins and treating digestive disorders | |
EP2236149B1 (en) | Medicinal composition for treating respiratory infectious diseases | |
WO2022206300A1 (en) | Composition capable of facilitating defecation and use thereof | |
CN112210516B (en) | Lactobacillus helveticus L1258 with intestinal regulation function and composition thereof | |
US10369184B2 (en) | Composition and use thereof in manufacture of product for improving intestinal function | |
CN105816623A (en) | Probiotic fermented traditional Chinese medicinal compound composition for treating and improving metabolic syndrome, and preparation method and application thereof | |
CN114344325A (en) | Application of bacteroides fragilis and zwitter-ion capsular polysaccharide thereof in preparation of medicines for preventing and treating genitourinary system tumors | |
CN113969253A (en) | Bifidobacterium lactis JYBR-390 with constipation treatment effect and application and product thereof | |
CN105663650A (en) | Probiotic fermentation traditional Chinese medicine composition for treating lung cancer and preparing method thereof | |
KR102480136B1 (en) | Lactobacillus paracasei with cognitive function improvement | |
CN109646546B (en) | A Chinese medicinal composition for preventing and treating diarrhea of livestock, and its preparation method | |
CN109700833B (en) | Pharmaceutical composition for treating constipation and preparation method and application thereof | |
CN109954004B (en) | Application of bacteroides fragilis extract in preparation of composition for preventing and treating psoriasis | |
CN114145460A (en) | Lactic acid bacteria, composition containing same and application | |
CN113116938A (en) | Quadruple viable bacteria preparation and application thereof | |
CN112972473A (en) | Medicine for treating adverse reactions of digestive tracts in radiotherapy and chemotherapy of tumors and preparation method thereof | |
CN105707898A (en) | Toxin removing and bowel relaxing probiotic composition and preparation method and application thereof | |
CN116790402B (en) | Bacteroides simplex strain with anti-inflammatory property, culture method and application | |
CN114158735B (en) | Probiotic composition and application thereof | |
CN116622593B (en) | Lactobacillus paracasei for fermentation and fermentation process for preparing wind-resistant acid-discharging ferment by same | |
CN114533750B (en) | Application of pericarpium Granati tannin in preparing medicine for treating enterotoxigenic escherichia coli intestinal diseases | |
CN109350661B (en) | Fermented traditional Chinese medicine composition for relieving cough and reducing sputum and application thereof in preventing and treating porcine respiratory diseases | |
CN109481474A (en) | Application of the bacteroides fragilis in the drug or food of preparation prevention and treatment rhinitis |
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 |