CN112481172B - 鼠李糖乳杆菌ccfm1130及其缓解治疗痛风的应用 - Google Patents
鼠李糖乳杆菌ccfm1130及其缓解治疗痛风的应用 Download PDFInfo
- Publication number
- CN112481172B CN112481172B CN202011486857.0A CN202011486857A CN112481172B CN 112481172 B CN112481172 B CN 112481172B CN 202011486857 A CN202011486857 A CN 202011486857A CN 112481172 B CN112481172 B CN 112481172B
- Authority
- CN
- China
- Prior art keywords
- lactobacillus rhamnosus
- ccfm1130
- serum
- hyperuricemia
- uric acid
- 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
- 241000218588 Lactobacillus rhamnosus Species 0.000 title claims abstract description 81
- 201000005569 Gout Diseases 0.000 title claims abstract description 16
- 201000001431 Hyperuricemia Diseases 0.000 claims abstract description 49
- 210000002966 serum Anatomy 0.000 claims abstract description 44
- 230000000694 effects Effects 0.000 claims abstract description 37
- LEHOTFFKMJEONL-UHFFFAOYSA-N Uric Acid Chemical compound N1C(=O)NC(=O)C2=C1NC(=O)N2 LEHOTFFKMJEONL-UHFFFAOYSA-N 0.000 claims abstract description 34
- TVWHNULVHGKJHS-UHFFFAOYSA-N Uric acid Natural products N1C(=O)NC(=O)C2NC(=O)NC21 TVWHNULVHGKJHS-UHFFFAOYSA-N 0.000 claims abstract description 33
- 229940116269 uric acid Drugs 0.000 claims abstract description 33
- 102100033220 Xanthine oxidase Human genes 0.000 claims abstract description 22
- 108010093894 Xanthine oxidase Proteins 0.000 claims abstract description 22
- 239000003814 drug Substances 0.000 claims abstract description 20
- HVYWMOMLDIMFJA-DPAQBDIFSA-N cholesterol Chemical compound C1C=C2C[C@@H](O)CC[C@]2(C)[C@@H]2[C@@H]1[C@@H]1CC[C@H]([C@H](C)CCCC(C)C)[C@@]1(C)CC2 HVYWMOMLDIMFJA-DPAQBDIFSA-N 0.000 claims abstract description 18
- 102000002260 Alkaline Phosphatase Human genes 0.000 claims abstract description 15
- 108020004774 Alkaline Phosphatase Proteins 0.000 claims abstract description 15
- WQZGKKKJIJFFOK-GASJEMHNSA-N Glucose Natural products OC[C@H]1OC(O)[C@H](O)[C@@H](O)[C@@H]1O WQZGKKKJIJFFOK-GASJEMHNSA-N 0.000 claims abstract description 9
- 239000008103 glucose Substances 0.000 claims abstract description 7
- 210000004369 blood Anatomy 0.000 claims description 16
- 239000008280 blood Substances 0.000 claims description 16
- LFQSCWFLJHTTHZ-UHFFFAOYSA-N Ethanol Chemical compound CCO LFQSCWFLJHTTHZ-UHFFFAOYSA-N 0.000 claims description 12
- 239000000203 mixture Substances 0.000 claims description 12
- 230000001580 bacterial effect Effects 0.000 claims description 7
- PEDCQBHIVMGVHV-UHFFFAOYSA-N Glycerine Chemical compound OCC(O)CO PEDCQBHIVMGVHV-UHFFFAOYSA-N 0.000 claims description 6
- 229920002472 Starch Polymers 0.000 claims description 6
- 239000008107 starch Substances 0.000 claims description 6
- 235000019698 starch Nutrition 0.000 claims description 6
- 150000003626 triacylglycerols Chemical class 0.000 claims description 5
- VYPSYNLAJGMNEJ-UHFFFAOYSA-N Silicium dioxide Chemical compound O=[Si]=O VYPSYNLAJGMNEJ-UHFFFAOYSA-N 0.000 claims description 4
- 239000007884 disintegrant Substances 0.000 claims description 4
- 239000003937 drug carrier Substances 0.000 claims description 4
- 239000000945 filler Substances 0.000 claims description 4
- 230000002401 inhibitory effect Effects 0.000 claims description 4
- 239000000314 lubricant Substances 0.000 claims description 4
- HQKMJHAJHXVSDF-UHFFFAOYSA-L magnesium stearate Chemical compound [Mg+2].CCCCCCCCCCCCCCCCCC([O-])=O.CCCCCCCCCCCCCCCCCC([O-])=O HQKMJHAJHXVSDF-UHFFFAOYSA-L 0.000 claims description 4
- 238000002360 preparation method Methods 0.000 claims description 4
- 239000000080 wetting agent Substances 0.000 claims description 4
- 230000000813 microbial effect Effects 0.000 claims description 3
- OWEGMIWEEQEYGQ-UHFFFAOYSA-N 100676-05-9 Natural products OC1C(O)C(O)C(CO)OC1OCC1C(O)C(O)C(O)C(OC2C(OC(O)C(O)C2O)CO)O1 OWEGMIWEEQEYGQ-UHFFFAOYSA-N 0.000 claims description 2
- GUBGYTABKSRVRQ-XLOQQCSPSA-N Alpha-Lactose Chemical compound O[C@@H]1[C@@H](O)[C@@H](O)[C@@H](CO)O[C@H]1O[C@@H]1[C@@H](CO)O[C@H](O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-XLOQQCSPSA-N 0.000 claims description 2
- FBPFZTCFMRRESA-KVTDHHQDSA-N D-Mannitol Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@H](O)CO FBPFZTCFMRRESA-KVTDHHQDSA-N 0.000 claims description 2
- 239000004375 Dextrin Substances 0.000 claims description 2
- 229920001353 Dextrin Polymers 0.000 claims 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 claims description 2
- GUBGYTABKSRVRQ-PICCSMPSSA-N Maltose Natural products O[C@@H]1[C@@H](O)[C@H](O)[C@@H](CO)O[C@@H]1O[C@@H]1[C@@H](CO)OC(O)[C@H](O)[C@H]1O GUBGYTABKSRVRQ-PICCSMPSSA-N 0.000 claims description 2
- 229930195725 Mannitol Natural products 0.000 claims description 2
- 229920000168 Microcrystalline cellulose Polymers 0.000 claims description 2
- WHNWPMSKXPGLAX-UHFFFAOYSA-N N-Vinyl-2-pyrrolidone Chemical compound C=CN1CCCC1=O WHNWPMSKXPGLAX-UHFFFAOYSA-N 0.000 claims description 2
- 239000000853 adhesive Substances 0.000 claims description 2
- 230000001070 adhesive effect Effects 0.000 claims description 2
- 239000011230 binding agent Substances 0.000 claims description 2
- 239000003795 chemical substances by application Substances 0.000 claims description 2
- 235000019425 dextrin Nutrition 0.000 claims description 2
- FPAFDBFIGPHWGO-UHFFFAOYSA-N dioxosilane;oxomagnesium;hydrate Chemical compound O.[Mg]=O.[Mg]=O.[Mg]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O.O=[Si]=O FPAFDBFIGPHWGO-UHFFFAOYSA-N 0.000 claims description 2
- 235000021107 fermented food Nutrition 0.000 claims description 2
- 230000006870 function Effects 0.000 claims description 2
- 235000013376 functional food Nutrition 0.000 claims description 2
- 235000012907 honey Nutrition 0.000 claims description 2
- 239000008101 lactose Substances 0.000 claims description 2
- 229940031703 low substituted hydroxypropyl cellulose Drugs 0.000 claims description 2
- 235000019359 magnesium stearate Nutrition 0.000 claims description 2
- 239000000594 mannitol Substances 0.000 claims description 2
- 235000010355 mannitol Nutrition 0.000 claims description 2
- 239000008108 microcrystalline cellulose Substances 0.000 claims description 2
- 235000019813 microcrystalline cellulose Nutrition 0.000 claims description 2
- 229940016286 microcrystalline cellulose Drugs 0.000 claims description 2
- 239000002417 nutraceutical Substances 0.000 claims description 2
- 235000021436 nutraceutical agent Nutrition 0.000 claims description 2
- 229920000036 polyvinylpyrrolidone Polymers 0.000 claims description 2
- 235000013855 polyvinylpyrrolidone Nutrition 0.000 claims description 2
- 229940069328 povidone Drugs 0.000 claims description 2
- 230000001737 promoting effect Effects 0.000 claims description 2
- 239000000377 silicon dioxide Substances 0.000 claims description 2
- 235000012239 silicon dioxide Nutrition 0.000 claims description 2
- 239000011734 sodium Substances 0.000 claims description 2
- 229910052708 sodium Inorganic materials 0.000 claims description 2
- OKODKVMXHLUQSW-JITBQSAISA-M sodium;(e)-4-hydroxy-4-oxobut-2-enoate;octadecanoic acid Chemical compound [Na+].OC(=O)\C=C\C([O-])=O.CCCCCCCCCCCCCCCCCC(O)=O OKODKVMXHLUQSW-JITBQSAISA-M 0.000 claims description 2
- 239000006188 syrup Substances 0.000 claims description 2
- 235000020357 syrup Nutrition 0.000 claims description 2
- 239000008194 pharmaceutical composition Substances 0.000 claims 3
- 210000004185 liver Anatomy 0.000 abstract description 11
- UFTFJSFQGQCHQW-UHFFFAOYSA-N triformin Chemical compound O=COCC(OC=O)COC=O UFTFJSFQGQCHQW-UHFFFAOYSA-N 0.000 abstract description 9
- 210000001035 gastrointestinal tract Anatomy 0.000 abstract description 8
- 229940079593 drug Drugs 0.000 abstract description 7
- 235000013305 food Nutrition 0.000 abstract description 4
- 244000005700 microbiome Species 0.000 abstract description 4
- 239000002068 microbial inoculum Substances 0.000 abstract description 2
- 230000002222 downregulating effect Effects 0.000 abstract 1
- 241000699670 Mus sp. Species 0.000 description 25
- 241000699666 Mus <mouse, genus> Species 0.000 description 15
- 108010090306 Member 2 Subfamily G ATP Binding Cassette Transporter Proteins 0.000 description 9
- 239000000243 solution Substances 0.000 description 9
- 241000124008 Mammalia Species 0.000 description 8
- 238000000034 method Methods 0.000 description 8
- 102100022595 Broad substrate specificity ATP-binding cassette transporter ABCG2 Human genes 0.000 description 7
- OFCNXPDARWKPPY-UHFFFAOYSA-N allopurinol Chemical compound OC1=NC=NC2=C1C=NN2 OFCNXPDARWKPPY-UHFFFAOYSA-N 0.000 description 7
- 238000010171 animal model Methods 0.000 description 7
- 230000000968 intestinal effect Effects 0.000 description 7
- 239000006041 probiotic Substances 0.000 description 7
- 235000018291 probiotics Nutrition 0.000 description 7
- 239000006228 supernatant Substances 0.000 description 7
- 238000011282 treatment Methods 0.000 description 7
- 229960003459 allopurinol Drugs 0.000 description 6
- 238000012258 culturing Methods 0.000 description 6
- 210000003405 ileum Anatomy 0.000 description 6
- 230000029142 excretion Effects 0.000 description 5
- 239000012530 fluid Substances 0.000 description 5
- 108020004999 messenger RNA Proteins 0.000 description 5
- 238000003672 processing method Methods 0.000 description 5
- 238000011160 research Methods 0.000 description 5
- 241000186660 Lactobacillus Species 0.000 description 4
- 238000001514 detection method Methods 0.000 description 4
- 206010012601 diabetes mellitus Diseases 0.000 description 4
- 230000002496 gastric effect Effects 0.000 description 4
- 239000001963 growth medium Substances 0.000 description 4
- 230000036541 health Effects 0.000 description 4
- 229940039696 lactobacillus Drugs 0.000 description 4
- 238000012163 sequencing technique Methods 0.000 description 4
- XLYOFNOQVPJJNP-UHFFFAOYSA-N water Chemical compound O XLYOFNOQVPJJNP-UHFFFAOYSA-N 0.000 description 4
- 241000894006 Bacteria Species 0.000 description 3
- 108010078791 Carrier Proteins Proteins 0.000 description 3
- 238000012449 Kunming mouse Methods 0.000 description 3
- 208000008589 Obesity Diseases 0.000 description 3
- 230000000052 comparative effect Effects 0.000 description 3
- 201000010099 disease Diseases 0.000 description 3
- 208000037265 diseases, disorders, signs and symptoms Diseases 0.000 description 3
- 238000002156 mixing Methods 0.000 description 3
- 235000020824 obesity Nutrition 0.000 description 3
- 230000001105 regulatory effect Effects 0.000 description 3
- 235000020183 skimmed milk Nutrition 0.000 description 3
- 230000004083 survival effect Effects 0.000 description 3
- 239000000725 suspension Substances 0.000 description 3
- 108020004465 16S ribosomal RNA Proteins 0.000 description 2
- 229920001817 Agar Polymers 0.000 description 2
- IJGRMHOSHXDMSA-UHFFFAOYSA-N Atomic nitrogen Chemical compound N#N IJGRMHOSHXDMSA-UHFFFAOYSA-N 0.000 description 2
- HEDRZPFGACZZDS-UHFFFAOYSA-N Chloroform Chemical compound ClC(Cl)Cl HEDRZPFGACZZDS-UHFFFAOYSA-N 0.000 description 2
- 102000004190 Enzymes Human genes 0.000 description 2
- 108090000790 Enzymes Proteins 0.000 description 2
- KFZMGEQAYNKOFK-UHFFFAOYSA-N Isopropanol Chemical compound CC(C)O KFZMGEQAYNKOFK-UHFFFAOYSA-N 0.000 description 2
- IAPCTXZQXAVYNG-UHFFFAOYSA-M Potassium 2,6-dihydroxytriazinecarboxylate Chemical compound [K+].[O-]C(=O)C1=NC(=O)NC(=O)N1 IAPCTXZQXAVYNG-UHFFFAOYSA-M 0.000 description 2
- 239000008272 agar Substances 0.000 description 2
- 238000004458 analytical method Methods 0.000 description 2
- 230000015572 biosynthetic process Effects 0.000 description 2
- 230000037396 body weight Effects 0.000 description 2
- 230000008859 change Effects 0.000 description 2
- YTRQFSDWAXHJCC-UHFFFAOYSA-N chloroform;phenol Chemical compound ClC(Cl)Cl.OC1=CC=CC=C1 YTRQFSDWAXHJCC-UHFFFAOYSA-N 0.000 description 2
- 235000012000 cholesterol Nutrition 0.000 description 2
- 239000012154 double-distilled water Substances 0.000 description 2
- 229940088598 enzyme Drugs 0.000 description 2
- 210000003608 fece Anatomy 0.000 description 2
- FDGQSTZJBFJUBT-UHFFFAOYSA-N hypoxanthine Chemical compound O=C1NC=NC2=C1NC=N2 FDGQSTZJBFJUBT-UHFFFAOYSA-N 0.000 description 2
- 230000006872 improvement Effects 0.000 description 2
- 230000037356 lipid metabolism Effects 0.000 description 2
- 230000007774 longterm Effects 0.000 description 2
- 239000006166 lysate Substances 0.000 description 2
- 239000002609 medium Substances 0.000 description 2
- 208000030159 metabolic disease Diseases 0.000 description 2
- 239000002207 metabolite Substances 0.000 description 2
- 239000002504 physiological saline solution Substances 0.000 description 2
- 238000004321 preservation Methods 0.000 description 2
- 239000000047 product Substances 0.000 description 2
- 230000002829 reductive effect Effects 0.000 description 2
- 238000005070 sampling Methods 0.000 description 2
- 238000012216 screening Methods 0.000 description 2
- 238000003786 synthesis reaction Methods 0.000 description 2
- 238000012360 testing method Methods 0.000 description 2
- XDIYNQZUNSSENW-UUBOPVPUSA-N (2R,3S,4R,5R)-2,3,4,5,6-pentahydroxyhexanal Chemical compound OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O.OC[C@@H](O)[C@@H](O)[C@H](O)[C@@H](O)C=O XDIYNQZUNSSENW-UUBOPVPUSA-N 0.000 description 1
- 108091032973 (ribonucleotides)n+m Proteins 0.000 description 1
- 201000001320 Atherosclerosis Diseases 0.000 description 1
- 241000304886 Bacilli Species 0.000 description 1
- 241000186000 Bifidobacterium Species 0.000 description 1
- 208000024172 Cardiovascular disease Diseases 0.000 description 1
- 102100035882 Catalase Human genes 0.000 description 1
- 108010053835 Catalase Proteins 0.000 description 1
- 230000004544 DNA amplification Effects 0.000 description 1
- 102100031181 Glyceraldehyde-3-phosphate dehydrogenase Human genes 0.000 description 1
- UGQMRVRMYYASKQ-UHFFFAOYSA-N Hypoxanthine nucleoside Natural products OC1C(O)C(CO)OC1N1C(NC=NC2=O)=C2N=C1 UGQMRVRMYYASKQ-UHFFFAOYSA-N 0.000 description 1
- 208000022559 Inflammatory bowel disease Diseases 0.000 description 1
- 241001465754 Metazoa Species 0.000 description 1
- 102000057297 Pepsin A Human genes 0.000 description 1
- 108090000284 Pepsin A Proteins 0.000 description 1
- ZLMJMSJWJFRBEC-UHFFFAOYSA-N Potassium Chemical compound [K] ZLMJMSJWJFRBEC-UHFFFAOYSA-N 0.000 description 1
- 238000011529 RT qPCR Methods 0.000 description 1
- FAPWRFPIFSIZLT-UHFFFAOYSA-M Sodium chloride Chemical compound [Na+].[Cl-] FAPWRFPIFSIZLT-UHFFFAOYSA-M 0.000 description 1
- 102100030935 Solute carrier family 2, facilitated glucose transporter member 9 Human genes 0.000 description 1
- 241001052560 Thallis Species 0.000 description 1
- 239000007983 Tris buffer Substances 0.000 description 1
- -1 Tris saturated phenol Chemical class 0.000 description 1
- 102000004142 Trypsin Human genes 0.000 description 1
- 108090000631 Trypsin Proteins 0.000 description 1
- 229940123769 Xanthine oxidase inhibitor Drugs 0.000 description 1
- 239000011543 agarose gel Substances 0.000 description 1
- 238000007605 air drying Methods 0.000 description 1
- 238000003556 assay Methods 0.000 description 1
- 230000004888 barrier function Effects 0.000 description 1
- 238000003287 bathing Methods 0.000 description 1
- 230000009286 beneficial effect Effects 0.000 description 1
- WHQCHUCQKNIQEC-UHFFFAOYSA-N benzbromarone Chemical compound CCC=1OC2=CC=CC=C2C=1C(=O)C1=CC(Br)=C(O)C(Br)=C1 WHQCHUCQKNIQEC-UHFFFAOYSA-N 0.000 description 1
- 229960002529 benzbromarone Drugs 0.000 description 1
- 239000003833 bile salt Substances 0.000 description 1
- 239000012620 biological material Substances 0.000 description 1
- 239000000872 buffer Substances 0.000 description 1
- 210000005252 bulbus oculi Anatomy 0.000 description 1
- 238000010804 cDNA synthesis Methods 0.000 description 1
- 208000026106 cerebrovascular disease Diseases 0.000 description 1
- 238000006243 chemical reaction Methods 0.000 description 1
- 230000001684 chronic effect Effects 0.000 description 1
- 239000002299 complementary DNA Substances 0.000 description 1
- 238000007796 conventional method Methods 0.000 description 1
- 230000007423 decrease Effects 0.000 description 1
- 230000008021 deposition Effects 0.000 description 1
- 235000005911 diet Nutrition 0.000 description 1
- 230000000378 dietary effect Effects 0.000 description 1
- 238000010790 dilution Methods 0.000 description 1
- 239000012895 dilution Substances 0.000 description 1
- 230000002526 effect on cardiovascular system Effects 0.000 description 1
- 239000002158 endotoxin Substances 0.000 description 1
- 230000003203 everyday effect Effects 0.000 description 1
- 238000002474 experimental method Methods 0.000 description 1
- 238000000605 extraction Methods 0.000 description 1
- 238000000855 fermentation Methods 0.000 description 1
- 230000004151 fermentation Effects 0.000 description 1
- 239000007850 fluorescent dye Substances 0.000 description 1
- 230000008014 freezing Effects 0.000 description 1
- 238000007710 freezing Methods 0.000 description 1
- 210000004051 gastric juice Anatomy 0.000 description 1
- 239000000499 gel Substances 0.000 description 1
- 108020004445 glyceraldehyde-3-phosphate dehydrogenase Proteins 0.000 description 1
- 230000012010 growth Effects 0.000 description 1
- 244000005709 gut microbiome Species 0.000 description 1
- 238000003384 imaging method Methods 0.000 description 1
- 238000000338 in vitro Methods 0.000 description 1
- 230000005764 inhibitory process Effects 0.000 description 1
- 239000007928 intraperitoneal injection Substances 0.000 description 1
- 230000006799 invasive growth in response to glucose limitation Effects 0.000 description 1
- PHTQWCKDNZKARW-UHFFFAOYSA-N isoamylol Chemical compound CC(C)CCO PHTQWCKDNZKARW-UHFFFAOYSA-N 0.000 description 1
- 210000003734 kidney Anatomy 0.000 description 1
- 208000017169 kidney disease Diseases 0.000 description 1
- 230000003907 kidney function Effects 0.000 description 1
- 239000007788 liquid Substances 0.000 description 1
- 239000012160 loading buffer Substances 0.000 description 1
- 238000011068 loading method Methods 0.000 description 1
- 238000004519 manufacturing process Methods 0.000 description 1
- 239000000463 material Substances 0.000 description 1
- 238000009629 microbiological culture Methods 0.000 description 1
- 238000012986 modification Methods 0.000 description 1
- 230000004048 modification Effects 0.000 description 1
- 239000006872 mrs medium Substances 0.000 description 1
- 229910052757 nitrogen Inorganic materials 0.000 description 1
- 231100000957 no side effect Toxicity 0.000 description 1
- 208000008338 non-alcoholic fatty liver disease Diseases 0.000 description 1
- 229960000988 nystatin Drugs 0.000 description 1
- VQOXZBDYSJBXMA-NQTDYLQESA-N nystatin A1 Chemical compound O[C@H]1[C@@H](N)[C@H](O)[C@@H](C)O[C@H]1O[C@H]1/C=C/C=C/C=C/C=C/CC/C=C/C=C/[C@H](C)[C@@H](O)[C@@H](C)[C@H](C)OC(=O)C[C@H](O)C[C@H](O)C[C@H](O)CC[C@@H](O)[C@H](O)C[C@](O)(C[C@H](O)[C@H]2C(O)=O)O[C@H]2C1 VQOXZBDYSJBXMA-NQTDYLQESA-N 0.000 description 1
- 229950000193 oteracil Drugs 0.000 description 1
- 238000000643 oven drying Methods 0.000 description 1
- 230000001575 pathological effect Effects 0.000 description 1
- 239000008188 pellet Substances 0.000 description 1
- 229960001412 pentobarbital Drugs 0.000 description 1
- WEXRUCMBJFQVBZ-UHFFFAOYSA-N pentobarbital Chemical compound CCCC(C)C1(CC)C(=O)NC(=O)NC1=O WEXRUCMBJFQVBZ-UHFFFAOYSA-N 0.000 description 1
- 229940111202 pepsin Drugs 0.000 description 1
- 239000011591 potassium Substances 0.000 description 1
- 229910052700 potassium Inorganic materials 0.000 description 1
- 239000002244 precipitate Substances 0.000 description 1
- 230000002265 prevention Effects 0.000 description 1
- 230000002062 proliferating effect Effects 0.000 description 1
- 238000010926 purge Methods 0.000 description 1
- 230000004144 purine metabolism Effects 0.000 description 1
- 230000009467 reduction Effects 0.000 description 1
- 230000000717 retained effect Effects 0.000 description 1
- 238000010839 reverse transcription Methods 0.000 description 1
- 230000002441 reversible effect Effects 0.000 description 1
- 238000000926 separation method Methods 0.000 description 1
- 238000004088 simulation Methods 0.000 description 1
- 239000008223 sterile water Substances 0.000 description 1
- 208000024891 symptom Diseases 0.000 description 1
- 230000002194 synthesizing effect Effects 0.000 description 1
- 231100000331 toxic Toxicity 0.000 description 1
- 230000002588 toxic effect Effects 0.000 description 1
- 239000012588 trypsin Substances 0.000 description 1
- 108010078530 urate transporter Proteins 0.000 description 1
- 230000003424 uricosuric effect Effects 0.000 description 1
- 238000005303 weighing Methods 0.000 description 1
- 238000012070 whole genome sequencing analysis Methods 0.000 description 1
- 239000003064 xanthine oxidase inhibitor Substances 0.000 description 1
Images
Classifications
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23L—FOODS, FOODSTUFFS, OR NON-ALCOHOLIC BEVERAGES, NOT COVERED BY SUBCLASSES A21D OR A23B-A23J; THEIR PREPARATION OR TREATMENT, e.g. COOKING, MODIFICATION OF NUTRITIVE QUALITIES, PHYSICAL TREATMENT; PRESERVATION OF FOODS OR FOODSTUFFS, IN GENERAL
- A23L33/00—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof
- A23L33/10—Modifying nutritive qualities of foods; Dietetic products; Preparation or treatment thereof using additives
- A23L33/135—Bacteria or derivatives thereof, e.g. probiotics
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61K—PREPARATIONS FOR MEDICAL, DENTAL OR TOILETRY PURPOSES
- A61K35/00—Medicinal preparations containing materials or reaction products thereof with undetermined constitution
- A61K35/66—Microorganisms or materials therefrom
- A61K35/74—Bacteria
- A61K35/741—Probiotics
- A61K35/744—Lactic acid bacteria, e.g. enterococci, pediococci, lactococci, streptococci or leuconostocs
- A61K35/747—Lactobacilli, e.g. L. acidophilus or L. brevis
-
- A—HUMAN NECESSITIES
- A61—MEDICAL OR VETERINARY SCIENCE; HYGIENE
- A61P—SPECIFIC THERAPEUTIC ACTIVITY OF CHEMICAL COMPOUNDS OR MEDICINAL PREPARATIONS
- A61P19/00—Drugs for skeletal disorders
- A61P19/06—Antigout agents, e.g. antihyperuricemic or uricosuric agents
-
- C—CHEMISTRY; METALLURGY
- C12—BIOCHEMISTRY; BEER; SPIRITS; WINE; VINEGAR; MICROBIOLOGY; ENZYMOLOGY; MUTATION OR GENETIC ENGINEERING
- C12N—MICROORGANISMS OR ENZYMES; COMPOSITIONS THEREOF; PROPAGATING, PRESERVING, OR MAINTAINING MICROORGANISMS; MUTATION OR GENETIC ENGINEERING; CULTURE MEDIA
- C12N1/00—Microorganisms, e.g. protozoa; Compositions thereof; Processes of propagating, maintaining or preserving microorganisms or compositions thereof; Processes of preparing or isolating a composition containing a microorganism; Culture media therefor
- C12N1/20—Bacteria; Culture media therefor
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2002/00—Food compositions, function of food ingredients or processes for food or foodstuffs
-
- A—HUMAN NECESSITIES
- A23—FOODS OR FOODSTUFFS; TREATMENT THEREOF, NOT COVERED BY OTHER CLASSES
- A23V—INDEXING SCHEME RELATING TO FOODS, FOODSTUFFS OR NON-ALCOHOLIC BEVERAGES AND LACTIC OR PROPIONIC ACID BACTERIA USED IN FOODSTUFFS OR FOOD PREPARATION
- A23V2400/00—Lactic or propionic acid bacteria
- A23V2400/11—Lactobacillus
- A23V2400/175—Rhamnosus
-
- Y—GENERAL TAGGING OF NEW TECHNOLOGICAL DEVELOPMENTS; GENERAL TAGGING OF CROSS-SECTIONAL TECHNOLOGIES SPANNING OVER SEVERAL SECTIONS OF THE IPC; TECHNICAL SUBJECTS COVERED BY FORMER USPC CROSS-REFERENCE ART COLLECTIONS [XRACs] AND DIGESTS
- Y02—TECHNOLOGIES OR APPLICATIONS FOR MITIGATION OR ADAPTATION AGAINST CLIMATE CHANGE
- Y02A—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE
- Y02A50/00—TECHNOLOGIES FOR ADAPTATION TO CLIMATE CHANGE in human health protection, e.g. against extreme weather
- Y02A50/30—Against vector-borne diseases, e.g. mosquito-borne, fly-borne, tick-borne or waterborne diseases whose impact is exacerbated by climate change
Landscapes
- Health & Medical Sciences (AREA)
- Life Sciences & Earth Sciences (AREA)
- Chemical & Material Sciences (AREA)
- Engineering & Computer Science (AREA)
- Organic Chemistry (AREA)
- Mycology (AREA)
- Microbiology (AREA)
- Bioinformatics & Cheminformatics (AREA)
- General Health & Medical Sciences (AREA)
- Medicinal Chemistry (AREA)
- Biotechnology (AREA)
- Genetics & Genomics (AREA)
- Pharmacology & Pharmacy (AREA)
- Zoology (AREA)
- Animal Behavior & Ethology (AREA)
- Wood Science & Technology (AREA)
- Public Health (AREA)
- Veterinary Medicine (AREA)
- Rheumatology (AREA)
- General Engineering & Computer Science (AREA)
- Virology (AREA)
- Biomedical Technology (AREA)
- Physical Education & Sports Medicine (AREA)
- Nuclear Medicine, Radiotherapy & Molecular Imaging (AREA)
- Chemical Kinetics & Catalysis (AREA)
- General Chemical & Material Sciences (AREA)
- Biochemistry (AREA)
- Tropical Medicine & Parasitology (AREA)
- Pain & Pain Management (AREA)
- Molecular Biology (AREA)
- Epidemiology (AREA)
- Nutrition Science (AREA)
- Food Science & Technology (AREA)
- Polymers & Plastics (AREA)
- Micro-Organisms Or Cultivation Processes Thereof (AREA)
- Medicines Containing Material From Animals Or Micro-Organisms (AREA)
- Coloring Foods And Improving Nutritive Qualities (AREA)
Abstract
本发明公开了鼠李糖乳杆菌CCFM1130及其缓解治疗痛风的应用,属于微生物技术领域。本发明的鼠李糖乳杆菌CCFM1130能够降低高尿酸血症小鼠的血清尿酸水平、抑制血清及肝脏的黄嘌呤氧化酶(XOD)活性,减少高尿酸血症和痛风的发生;下调血清血糖、下调血清总甘油三酯(TG)、下调血清总胆固醇(TC)水平及抑制血清碱性磷酸酶(ALP)活性;并促进回肠尿酸转运蛋白ABCG2的表达。本发明的鼠李糖乳杆菌CCFM1130能够耐受胃肠道环境,可用于制备缓解高尿酸血症及痛风的可摄入胃肠道的功能性菌剂、食品和药物,具有广泛的应用前景。
Description
技术领域
本发明涉及鼠李糖乳杆菌CCFM1130及其缓解治疗痛风的应用,属于微生物技术领域。
背景技术
高尿酸血症(Hyperuricemia,HUA)是指血液内尿酸水平超过正常值的一种疾病。近年来,随着生活水平的提高,高尿酸血症的发病率也越来越高,我国高尿酸血症患者占总人口约13.3%。长期的高尿酸血症造成的尿酸盐结石会进一步诱发痛风。同时,高尿酸血症被认为是心脑血管疾病、慢性肾病、动脉粥样硬化的危险因素,严重危害人类健康,因此高尿酸血症的治疗引起了人们的高度重视。目前治疗高尿酸血症的药物主要有别嘌呤醇(黄嘌呤氧化酶抑制剂)、苯溴马隆(促尿酸排泄药物)等,但是这些药物存在一些副作用,国际上针对无症状高尿酸血症的降尿酸药物治疗存在诸多争议。因此,在饮食及生活方式上进行改善是无症状高尿酸血症治疗的优选方法。
近年来,随着对肠道菌群与人体健康关系的深入研究,大量研究证实了益生菌通过调节肠道菌群对人体健康的改善功能。高尿酸血症的发病与肠道菌群结构紊乱密切相关,而食入益生菌可通过增殖肠道中的乳杆菌和双歧杆菌调节肠道微生物群,改善肠道屏障功能,减少内毒素等代谢物随血液进入肝脏,有效降低血尿酸值。尿酸在人体内的排泄主要通过肾脏及肠道的排泄作用,ABCG2作为尿酸转运蛋白在尿酸的肠排泄中发挥着重要作用,肠道ABCG2 的表达被认为是治疗高尿酸血症及痛风的新靶点,但目前针对肠道ABCG2的药物尚未有发现。
益生菌具有安全、无副作用等特点,目前已有大量临床及动物实验研究表明,益生菌具有缓解肥胖、非酒精性脂肪肝、炎症性肠病等作用。随着对益生菌研究的不断深入,通过益生菌来改善高尿酸血症患者的健康状况,成为高尿酸血症治疗及预防痛风发生的新手段。
发明内容
因此,作为本发明其中一个方面,本发明克服现有技术中存在的不足提供了一种鼠李糖乳杆菌(Lactobacillus rhamnosus)CCFM1130,已于2020年7月22日保藏于广东省微生物菌种保藏中心,地址为广东省广州市先烈中路100号大院59号楼5楼,保藏编号为GDMCCNo:61091。
该菌株具有如下特征:
(1)菌体特征:呈革兰氏染色阳性,不形成孢子,非运动性的细菌。
(2)菌落特征:呈乳白色、有光泽、凸起、不透明、光滑整齐。
(3)生长特性:在37℃恒温条件下,在MRS培养基培养约12h到达对数末期。
(4)对模拟胃肠液具有较强耐受能力。
本发明的第二个目的是提供含有所述鼠李糖乳杆菌的组合物。
在一种实施方式中,所述鼠李糖乳杆菌CCFM1130的数量≥1×106CFU/mL或≥1×106CFU/g。
在一种实施方式中,所述鼠李糖乳杆菌CCFM1130的数量≥1×109CFU/mL或≥1×109CFU/g。
在一种实施方式中,所述组合物包括但不限于微生物制剂、功能性食品、保健品或药物。
在一种实施方式中,所述组合物中含有所述鼠李糖乳杆菌CCFM1130的活菌株、干菌株、菌株代谢物或灭活的菌株。
在一种实施方式中,所述药物具有如下至少一种作用:
(1)降低高尿酸血症哺乳动物的血清尿酸水平;
(2)降低高尿酸血症哺乳动物的血清及肝脏黄嘌呤氧化酶(XOD)活性;
(3)降低哺乳动物的血糖水平;
(4)降低哺乳动物血清总甘油三酯水平;
(5)降低哺乳动物的血清总胆固醇水平;
(6)降低哺乳动物的血清碱性磷酸酶的活性;
(7)提高哺乳动物回肠尿酸转运蛋白ABCG2的mRNA水平。
在一种实施方式中,所述哺乳动物包括但不限于人类。
在一种实施方式中,所述药物还含有药学上可接受的载体。
在一种实施方式中,所述药学上可接受的载体包括但不限于:填充剂、润湿剂、崩解剂、粘合剂或润滑剂中的一种或多种。
在一种实施方式中,所述填充剂为微晶纤维素、乳糖、甘露醇、淀粉或糊精中的一种或多种;所述润湿剂为乙醇或甘油中的一种或多种;所述崩解剂为羧甲基淀粉钠、交联羧甲基淀粉钠、交联聚维酮或低取代羟丙基纤维素中的一种或多种;所述粘合剂为淀粉糊、糖浆、饴糖、炼蜜或液状葡萄糖中的一种或多种;所述润滑剂为硬脂酸镁、硬脂酸富马酸钠、滑石粉或二氧化硅中的一种或多种。
本发明的第三个目的是提供所述鼠李糖乳杆菌CCFM1130在制备缓解高尿酸血症和痛风的功能性菌剂、食品或药物中的应用。
在一种实施方式中,所述药物用于降低高尿酸血症哺乳动物的血清尿酸水平。
在一种实施方式中,所述药物用于抑制黄嘌呤氧化酶活性。
在一种实施方式中,所述药物用于调节血糖升高。
在一种实施方式中,所述药物用于制备调节血清总甘油三酯和/或血清总胆固醇升高。
在一种实施方式中,所述药物用于调节血清碱性磷酸酶活性升高。
在一种实施方式中,所述药物用于促进回肠尿酸转运蛋白ABCG2表达。
本发明还要求保护所述鼠李糖乳杆菌CCFM1130在制备发酵食品中的应用。
在一种实施方式中,所述应用包括但不限于将所述鼠李糖乳杆菌CCFM1130作为发酵微生物,利用食品原料进行发酵。
本发明的有益效果:所述鼠李糖乳杆菌CCFM1130能够耐受胃肠道环境,可用于制备缓解高尿酸血症及痛风的功能性菌剂、食品和药物;所述鼠李糖乳杆菌CCFM1130能够降低血清尿酸水平,降低血清及肝脏的黄嘌呤氧化酶(XOD)活性,减少痛风的发生;所述鼠李糖乳杆菌CCFM1130能够下调血清血糖、下调血清总甘油三酯(TG)、下调血清总胆固醇(TC)水平及抑制血清碱性磷酸酶(ALP)活性;此外,所述鼠李糖乳杆菌CCFM1130能够促进回肠尿酸转运蛋白ABCG2的表达。具有广泛的应用前景。
生物材料保藏
鼠李糖乳杆菌CCFM1130,分类命名为Lactobacillus rhamnosus,已于2020年7月22日保藏于广东省微生物菌种保藏中心,保藏编号为GDMCC No:61091。
附图说明
图1是鼠李糖乳杆菌CCFM1130的菌落形态;
图2是鼠李糖乳杆菌CCFM1130对高尿酸血症小鼠血清尿酸的影响;
图3是鼠李糖乳杆菌CCFM1130对高尿酸血症小鼠血清和肝脏黄嘌呤氧化酶(XOD)活力的影响;
图4是鼠李糖乳杆菌CCFM1130对高尿酸血症小鼠血糖(Glucose)的影响;
图5是鼠李糖乳杆菌CCFM1130对高尿酸血症小鼠总甘油三酯(TG)的影响;
图6是鼠李糖乳杆菌CCFM1130对高尿酸血症小鼠血清总胆固醇(TC)的影响;
图7是鼠李糖乳杆菌CCFM1130对高尿酸血症小鼠血清碱性磷酸酶(ALP)的影响;
图8是鼠李糖乳杆菌CCFM1130对高尿酸血症小鼠回肠尿酸转运蛋白ABCG2 mRNA水平的影响;
其中,*P<0.05,**P<0.01,***P<0.001,****P<0.0001(与高尿酸血症模型组比较)。
具体实施方式
实施例1鼠李糖乳杆菌CCFM1130的筛选
(一)乳杆菌的分离筛选
(1)取1g健康成年人的新鲜粪便。梯度稀释后涂于加有1%制霉菌素的LBS培养基,置于37℃恒温培养箱中培养48h。
(2)培养后根据菌落的颜色、大小、边缘形状等,用接种环挑取菌落划线纯化。
(3)所得菌落进行革兰氏染色和过氧化氢酶分析。
(4)保留革兰氏染色阳性杆菌和过氧化氢酶阴性菌。
(二)乳杆菌的分子生物学鉴定
(1)单菌基因组抽提
(A)将步骤(一)所筛选得到的乳杆菌培养过夜;
(B)取培养过夜的菌悬液lmL于1.5mL离心管,10000r/min离心2min,弃上清得菌体;
(C)用lmL无菌水吹洗菌体后,10000r/min离心2min,弃上清得菌体;
(D)加入200μL SDS裂解液,80℃水浴30min;
(E)加入酚-氯仿溶液200μL于菌体裂解液中,其中酚-氯仿溶液的组成成分及体积比为Tris饱和酚:氯仿:异戊醇=25:24:1,颠倒混匀后,12000rpm离心5-10min,取上清200μL;
(F)加入400μL冰乙醇或冰异丙醇于200μL上清中,-20℃静置1h,12000rpm离心 5-10min,弃上清;
(G)加入500μL70%(体积百分数)冰乙醇重悬沉淀,12000rpm离心1-3min,弃上清;60℃烘箱烘干,或者自然晾干;
(H)50μL ddH2O重溶沉淀以备PCR。
(2)16S rDNA PCR
(A)细菌16S rDNA 50μLPCR反应体系
10×Taq buffer,5μL;dNTP,5μL;引物27F,0.5μL;引物1492R,0.5μL;Taq酶,0.5μL;模板,0.5μL;ddH2O,38μL。
(B)PCR条件
95℃5min;95℃10s;55℃30s;72℃30s;step2-4 30×;72℃5min;12℃2min。
(C)制备1%琼脂糖凝胶,之后将PCR产物与10000×loading buffer混合,上样量2μL, 120V跑30min,然后进行凝胶成像;
(D)将得到PCR产物送专业测序公司,将得到的测序结果与使用BLAST在GenBank中进行搜索和相似性比对,将鉴定为鼠李糖乳杆菌的菌株-80℃保存。
(3)全基因组测序
将提取的全基因组送专业测序公司,利用二代测序仪对菌的全基因组进行测序,将得到的序列结果使用BLAST在GenBank中进行搜索和相似性比对,测序结果鉴定为属于鼠李糖乳杆菌的一种新发现的菌株-80℃保藏备用。
实施例2:鼠李糖乳杆菌CCFM1130对模拟胃肠液的耐受性检测
将冷冻保存的鼠李糖乳杆菌CCFM1130接种于MRS培养基中,在温度37℃条件下培养 14h,再经MRS培养液传代培养2~3次后。
取3mL鼠李糖乳杆菌CCFM1130的培养液8000×g离心2min收集菌体,与3mL pH 3.0人工模拟胃液(含3g/L胃蛋白酶、pH=3.0的生理盐水)混合,并在37℃培养箱培养,分别在0h、2h时取样,用MRS琼脂培养基浇注培养进行平板菌落计数,测定活菌数并计算其存活率。
取3mL鼠李糖乳杆菌CCFM1130的培养液8000×g离心2min收集菌体,加入3mL pH8.0 人工模拟肠液(含胰蛋白酶1g/L、胆盐0.3%、pH=8.0的生理盐水)混合,于37℃培养,分别在0h、2h、4h取样,用MRS琼脂培养基浇注培养进行平板菌落计数,测定活菌数并计算其存活率。
存活率(%)以该培养液中在取样时的活菌数与在第0h时活菌数之比计算。实验结果如表1所示,结果表明,鼠李糖乳杆菌CCFM1130对人工模拟胃肠液具有较好的耐受能力。
表1 鼠李糖乳杆菌CCFM1130在人工模拟胃肠液中的耐受能力
实施例3:鼠李糖乳杆菌CCFM1130对KunMing小鼠无毒副作用
将鼠李糖乳杆菌CCFM1130菌体重悬于100g/L的脱脂乳溶液中,制成浓度为 4.0×109CFU/mL的菌悬液。取体重24-32g左右的健康雄性KunMing小鼠12只,适应环境一周后,分为CCFM1130组和对照组。CCFM1130组每日给予该浓度菌悬液0.3mL灌胃一次,对照组灌胃相同体积的不含鼠李糖乳杆菌CCFM1130的100g/L的脱脂乳溶液,观察一周,记录死亡和体重情况。
这些试验结果列于表2中。结果表明,喂食浓度1×109CFU/只的鼠李糖乳杆菌CCFM1130 未对小鼠造成明显影响,体重无显著变化,无死亡现象产生。小鼠外观无明显病理症状。
表2 小鼠体重变化及死亡情况
实施例4:鼠李糖乳杆菌CCFM1130能够降低高尿酸血症小鼠的血清尿酸水平
取体重24-32g的健康雄性KunMing小鼠24只,适应性培养1周后,随机分为4组,分别为对照组、高尿酸血症模型组、鼠李糖乳杆菌CCFM1130干预组(CCFM1130)和别嘌呤醇干预组(别嘌呤醇)。除对照组外,其余组每天灌胃500mg/kg BW次黄嘌呤,1h后腹腔注射200mg/kg BW氧嗪酸钾;在氧嗪酸钾处理前1h,对照组和高尿酸血症模型组给予100g/L 脱脂乳,鼠李糖乳杆菌CCFM1130干预组给予1.0×109CFU/只的鼠李糖乳杆菌CCFM1130,别嘌呤醇组给予5mg/kg BW别嘌呤醇。实验分组及处理方法见表3:
表3 实验动物分组情况
实验末期收集小鼠新鲜粪便冻存于-80℃。试验结束时,小鼠禁食不禁水12h,腹腔注射 0.1mL/10g 1%的戊巴比妥钠溶液麻醉后,摘眼球取血并辅以颈椎脱臼法处死。血液样本 3500r/min离心15分钟,取上清,-80℃冻存,用于血液指标分析。肝脏、回肠等组织取出后迅速于预冷的生理盐水中漂洗去血,于液氮中速冻并转移于-80℃冻存,后续制成肝匀浆测定相关指标。血清尿酸水平的测定按照试剂盒方法进行。
鼠李糖乳杆菌CCFM1130对小鼠血清尿酸水平的影响如图2,与高尿酸血症模型小鼠相比,鼠李糖乳杆菌CCFM1130能够将高尿酸血症模型小鼠的血清尿酸水平降低22.78%,并接近对照组,其降低尿酸的作用与药物别嘌呤醇相近,可预防和减少高尿酸血症和痛风的发生。
实施例5:鼠李糖乳杆菌CCFM1130能够降低高尿酸血症小鼠的黄嘌呤氧化酶活性
实验动物分组及处理方法同实施例4,黄嘌呤氧化酶(XOD)的检测按照试剂盒(北京索莱宝)的方法进行。
黄嘌呤氧化酶是嘌呤代谢合成尿酸的关键酶,降尿酸药物别嘌呤醇由于能够抑制黄嘌呤氧化酶的活性使得尿酸合成减少,从而发挥降尿酸作用。如图3所示,与高尿酸血症模型小鼠相比,鼠李糖乳杆菌CCFM1130能够降低31.37%高尿酸血症模型小鼠的肝脏黄嘌呤氧化酶活性和37.63%的血清黄嘌呤氧化酶活性,鼠李糖乳杆菌CCFM1130使得高尿酸血症小鼠的血清和肝脏黄嘌呤氧化酶活力升高趋近正常,从而使小鼠体内尿酸合成减少,有利于高尿酸血症和痛风的预防和治疗。
实施例6:鼠李糖乳杆菌CCFM1130下调血糖水平
实验动物分组及处理方法同实施例4,血糖的检测使用迈瑞BS480生化分析仪按照试剂盒的方法进行。
大量研究表明,糖尿病与高尿酸血症作为代谢性疾病,存在着各种各样的联系。长期的糖尿病引起的肾脏功能下降会使血清尿酸升高,引起高尿酸血症甚至痛风的发生,而高尿酸血症的发生也增加了糖尿病的风险。血糖结果显示(图4),高尿酸血症小鼠血糖浓度达 4.52±0.58mmol/L,而鼠李糖乳杆菌CCFM1130能够使得小鼠的血糖降至正常值 2.42±0.76mmol/L,这说明鼠李糖乳杆菌CCFM1130具备缓解高尿酸血症及糖尿病等代谢性疾病的潜力。
实施例7:鼠李糖乳杆菌CCFM1130下调血清总甘油三酯水平
实验动物分组及处理方法同实施例4,血清总甘油三酯(TG)的检测使用迈瑞BS480生化分析仪按照试剂盒的方法进行。
鼠李糖乳杆菌CCFM1130对高尿酸血症小鼠的血清总甘油三酯的影响如图5,与对照组相比,高尿酸血症小鼠具有较高的血清总甘油三酯浓度,达1.41±0.27mmol/L,鼠李糖乳杆菌 CCFM1130能够使其恢复至正常水平1.03±0.15mmol/L。这说明鼠李糖乳杆菌CCFM1130具备调节脂质代谢,缓解肥胖等疾病的潜力。
实施例8:鼠李糖乳杆菌CCFM1130下调血清总胆固醇水平
实验动物分组及处理方法同实施例4,血清总胆固醇(TC)的检测使用迈瑞BS480生化分析仪按照试剂盒的方法进行。
鼠李糖乳杆菌CCFM1130对高尿酸血症小鼠的血清总胆固醇的影响如图6,与对照组相比,高尿酸血症小鼠具有较高的血清总胆固醇水平,胆固醇含量达2.90±0.37mmol/L,鼠李糖乳杆菌CCFM1130能够使其恢复至与对照组相当的正常水平2.31±0.34mmol/L。这说明鼠李糖乳杆菌CCFM1130具备调节脂质代谢,缓解肥胖等疾病的潜力。
实施例9:鼠李糖乳杆菌CCFM1130降低小鼠的血清碱性磷酸酶(ALP)的活性
实验动物分组及处理方法同实施例4,血清碱性磷酸酶(ALP)的检测使用迈瑞BS480 生化分析仪按照试剂盒的方法进行。
结果显示(图7),与对照组相比,高尿酸血症小鼠血清碱性磷酸酶活性升高至86.5±14.0U/L,而鼠李糖乳杆菌CCFM1130干预能够使得升高的碱性磷酸酶活性降低至51.6±7.5U/L。
实施例10:鼠李糖乳杆菌CCFM1130提高小鼠回肠尿酸转运蛋白ABCG2的mRNA水平
实验动物分组及处理方法同实施例4。回肠ABCG2 mRNA测定:取约20mg回肠组织加入500μL Trizol,冰浴匀浆后采用常规方法提取回肠组织中的RNA。按照反转录试剂盒说明书进行cDNA合成。用荧光染料SYBR Green super mix(Qiagen,Germany)混合样本,PCR 体系为5μL mix、1μL cDNA、1μL正向和1μL反向引物,用ddH2O补至总体积为10μL。在实时荧光定量基因扩增仪器CFX96TM Real-Time System(Bio-Rad,USA)上进行检测,每个样本设立3个平行孔,并以GAPDH为内参,所得结果用2-ΔΔCq的方法进行分析;所用的引物序列见表4。
表4 qPCR引物序列
结果显示(图8),鼠李糖乳杆菌CCFM1130能够显著提高高尿酸血症小鼠的回肠ABCG2 的mRNA水平。回肠ABCG2在肠道尿酸排泄中发挥着重要作用,鼠李糖乳杆菌CCFM1130能够通过提高回肠ABCG2的表达从而促进尿酸排出体外。
对比例1:
具体实施方式同实施例4,区别在于,将鼠李糖乳杆菌CCFM1130替换为鼠李糖乳杆菌FSHMX12(公开于:黄丹.人肠道鼠李糖乳杆菌的比较基因组与部分生理生化特性研究[D]. 江南大学,2019),测定小鼠的血清尿酸指标,结果显示,鼠李糖乳杆菌FSHMX12组小鼠的血清尿酸值为611.9±68.0μmol/L,相比于高尿酸血症模型组(623.0±76.7μmol/L)未发生明显变化。
对比例2
具体实施方式同实施例5,区别在于,将鼠李糖乳杆菌CCFM1130替换为鼠李糖乳杆菌 FSHMX12,测定高尿酸血症小鼠的黄嘌呤氧化酶活性,结果显示鼠李糖乳杆菌FSHMX12组小鼠的血清和肝脏黄嘌呤氧化酶活性分别为7.832±0.988U/L、2.124±0.307U/g prot,相比于高尿酸血症模型组(7.531±1.440U/L、1.718±0.416U/g prot),鼠李糖乳杆菌FSHMX12未对高尿酸血症小鼠的黄嘌呤氧化酶活性起到明显抑制作用,反而使得高尿酸血症小鼠血清和肝脏黄嘌呤氧化酶活性有所升高。
虽然本发明已以较佳实施例公开如上,但其并非用以限定本发明,任何熟悉此技术的人,在不脱离本发明的精神和范围内,都可做各种的改动与修饰,因此本发明的保护范围应该以权利要求书所界定的为准。
Claims (10)
1.鼠李糖乳杆菌(Lactobacillus rhamnosus)CCFM1130,已于2020年7月22日保藏于广东省微生物菌种保藏中心,保藏编号为GDMCC No:61091。
2.含有权利要求1所述鼠李糖乳杆菌CCFM1130的组合物。
3.根据权利要求2所述的组合物,其特征在于,所述组合物为微生物制剂、功能性食品、保健品或药物。
4.根据权利要求2或3所述的组合物,其特征在于,所述组合物中含有所述鼠李糖乳杆菌CCFM1130的活菌株、干菌株或灭活的菌株。
5.一种药物组合物,其特征在于,含有权利要求1所述的鼠李糖乳杆菌CCFM1130和药学上可接受的载体。
6.根据权利要求5所述的药物组合物,其特征在于,所述药学上可接受的载体包括但不限于:填充剂、润湿剂、崩解剂、粘合剂或润滑剂中的一种或多种。
7.根据权利要求6所述的药物组合物,其特征在于,所述填充剂为微晶纤维素、乳糖、甘露醇、淀粉或糊精中的一种或多种;所述润湿剂为乙醇或甘油中的一种或多种;所述崩解剂为羧甲基淀粉钠、交联羧甲基淀粉钠、交联聚维酮或低取代羟丙基纤维素中的一种或多种;所述粘合剂为淀粉糊、糖浆、饴糖、炼蜜或液状葡萄糖中的一种或多种;所述润滑剂为硬脂酸镁、硬脂酸富马酸钠、滑石粉或二氧化硅中的一种或多种。
8.权利要求1所述的鼠李糖乳杆菌CCFM1130在制备缓解高尿酸血症和痛风的功能性菌剂或药物中的应用。
9.根据权利要求8所述的应用,其特征在于,具有如下至少一种功能:
(1)抑制黄嘌呤氧化酶活性;
(2)抑制血糖升高;
(3)调节血清总甘油三酯和/或血清总胆固醇;
(4)抑制血清碱性磷酸酶升高;
(5)促进回肠尿酸转运蛋白ABCG2表达。
10.权利要求1所述的鼠李糖乳杆菌CCFM1130在制备发酵食品中的应用。
Priority Applications (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011486857.0A CN112481172B (zh) | 2020-12-16 | 2020-12-16 | 鼠李糖乳杆菌ccfm1130及其缓解治疗痛风的应用 |
Applications Claiming Priority (1)
Application Number | Priority Date | Filing Date | Title |
---|---|---|---|
CN202011486857.0A CN112481172B (zh) | 2020-12-16 | 2020-12-16 | 鼠李糖乳杆菌ccfm1130及其缓解治疗痛风的应用 |
Publications (2)
Publication Number | Publication Date |
---|---|
CN112481172A CN112481172A (zh) | 2021-03-12 |
CN112481172B true CN112481172B (zh) | 2022-07-05 |
Family
ID=74917244
Family Applications (1)
Application Number | Title | Priority Date | Filing Date |
---|---|---|---|
CN202011486857.0A Active CN112481172B (zh) | 2020-12-16 | 2020-12-16 | 鼠李糖乳杆菌ccfm1130及其缓解治疗痛风的应用 |
Country Status (1)
Country | Link |
---|---|
CN (1) | CN112481172B (zh) |
Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108486007A (zh) * | 2018-03-22 | 2018-09-04 | 嘉兴益诺康生物科技有限公司 | 一种用于降低血尿酸的干酪乳杆菌株、益生菌组合物及其应用 |
CN112263669A (zh) * | 2020-09-21 | 2021-01-26 | 康美华大基因技术有限公司 | 一种用于缓解高尿酸血症的低聚肽中药益生菌复合物 |
CN112458027A (zh) * | 2020-12-16 | 2021-03-09 | 江南大学 | 一株格氏乳杆菌及其缓解和治疗高尿酸血症的用途 |
CN112574917A (zh) * | 2020-12-16 | 2021-03-30 | 江南大学 | 一株缓解高尿酸血症的鼠李糖乳杆菌ccfm1131 |
Family Cites Families (5)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN104887716A (zh) * | 2014-03-07 | 2015-09-09 | 景岳生物科技股份有限公司 | 用于治疗第二型糖尿病的罗伊氏乳杆菌gmnl-89组合物及其用途 |
US9636368B2 (en) * | 2014-08-22 | 2017-05-02 | Food Industry Research And Development Institute | Strain of Lactobacillus rhamnosus and its metabolites for use in inhibiting xanthine oxidase and treating gout |
WO2018204764A1 (en) * | 2017-05-05 | 2018-11-08 | Camp4 Therapeutics Corporation | Identification and targeted modulation of gene signaling networks |
WO2019203827A1 (en) * | 2018-04-19 | 2019-10-24 | Kibow Biotech Inc . | Composition and method for preventing or treating hyperuricemia or gout |
CN111826315B (zh) * | 2020-07-20 | 2022-04-01 | 广东南芯医疗科技有限公司 | 一株鼠李糖乳杆菌nx-2及其在制备降尿酸药物中的应用 |
-
2020
- 2020-12-16 CN CN202011486857.0A patent/CN112481172B/zh active Active
Patent Citations (4)
Publication number | Priority date | Publication date | Assignee | Title |
---|---|---|---|---|
CN108486007A (zh) * | 2018-03-22 | 2018-09-04 | 嘉兴益诺康生物科技有限公司 | 一种用于降低血尿酸的干酪乳杆菌株、益生菌组合物及其应用 |
CN112263669A (zh) * | 2020-09-21 | 2021-01-26 | 康美华大基因技术有限公司 | 一种用于缓解高尿酸血症的低聚肽中药益生菌复合物 |
CN112458027A (zh) * | 2020-12-16 | 2021-03-09 | 江南大学 | 一株格氏乳杆菌及其缓解和治疗高尿酸血症的用途 |
CN112574917A (zh) * | 2020-12-16 | 2021-03-30 | 江南大学 | 一株缓解高尿酸血症的鼠李糖乳杆菌ccfm1131 |
Non-Patent Citations (1)
Title |
---|
食品中嘌呤的降低方法及低嘌呤产品研究进展;刘建林等;《食品研究与开发》;20200120(第02期);全文 * |
Also Published As
Publication number | Publication date |
---|---|
CN112481172A (zh) | 2021-03-12 |
Similar Documents
Publication | Publication Date | Title |
---|---|---|
CN112458027B (zh) | 一株格氏乳杆菌及其缓解和治疗高尿酸血症的用途 | |
CN112574917B (zh) | 一株缓解高尿酸血症的鼠李糖乳杆菌ccfm1131 | |
CN114196581B (zh) | 缓解高尿酸血症的罗伊氏乳杆菌ccfm1132及应用 | |
CN112813003B (zh) | 一株植物乳杆菌及其在制备缓解高血脂症引发的疾病的药物或食品中的应用 | |
CN113604384B (zh) | 一种鼠李糖乳杆菌及其应用 | |
CN111560330B (zh) | 一种具有免疫调节、抗炎和抗宫颈癌作用的干酪乳杆菌及应用 | |
CN114181864B (zh) | 一种鼠李糖乳杆菌hf01及其应用 | |
CN110368406B (zh) | 青春双歧杆菌ccfm1062在制备功能性菌剂、食品和/或药物中的应用 | |
CN109593678B (zh) | 长双歧杆菌yh295及其在制备降低腹型肥胖风险产品中的应用 | |
CN114574390A (zh) | 一株缓解结肠炎的长双歧杆菌婴儿亚种及应用 | |
CN113025526B (zh) | 一株减少结肠病理损伤且具有缓解便秘作用的两歧双歧杆菌 | |
WO2021143621A1 (zh) | Anaerostipes sp B2131菌及其在炎症性肠病中的应用 | |
CN115287240A (zh) | 一株具有高尿酸血症及痛风防治作用的植物乳杆菌及其应用 | |
CN114107088B (zh) | 一种罗伊氏乳杆菌lrsy523及其应用 | |
CN113797232B (zh) | 具有缓解胰岛素抵抗功能的组合物及其应用 | |
CN114854638A (zh) | 一株高效表达腺苷脱氨酶缓解结肠炎副干酪乳杆菌 | |
CN113151070B (zh) | 一株可提高肠道中克里斯藤森菌科相对丰度的发酵乳杆菌 | |
CN110331118B (zh) | 青春双歧杆菌ccfm1061、其发酵食品及菌剂制备方法 | |
CN116555074B (zh) | 一株短乳杆菌jt1及其在制备降血糖药品中的应用 | |
CN112481172B (zh) | 鼠李糖乳杆菌ccfm1130及其缓解治疗痛风的应用 | |
CN113249256B (zh) | 一株缓解雌激素相关代谢紊乱及肥胖的植物乳杆菌及应用 | |
CN116769658A (zh) | 一种凝结芽孢杆菌ap15及其在肠道调节与尿酸、胆固醇代谢的应用 | |
CN114410532B (zh) | 一株降低血浆氧化三甲胺和盲肠三甲胺水平的长双歧杆菌及其应用 | |
CN111700918B (zh) | 一种用于缓解酒精性肠道损伤的药品 | |
CN114410531A (zh) | 降低血浆tmao及缓解、预防动脉粥样硬化的长双歧杆菌ccfm1216及其应用 |
Legal Events
Date | Code | Title | Description |
---|---|---|---|
PB01 | Publication | ||
PB01 | Publication | ||
SE01 | Entry into force of request for substantive examination | ||
SE01 | Entry into force of request for substantive examination | ||
GR01 | Patent grant | ||
GR01 | Patent grant |