微囊化益生菌对小鼠急性溃疡性结肠炎的影响

doi:10.16431/j.cnki.1671-7236.2020.08.026

摘要/Abstract

摘要： 试验旨在观察微囊化布拉迪酵母菌（Saccharomyces boulardii，S.boulardii）与微囊化粪肠球菌（Enterococcus faecium，E.faecalis）对小鼠溃疡性结肠炎（UC）的治疗效果并探讨其作用机制。采用3%葡聚糖硫酸钠（dextran sulfacte sodium，DSS）灌胃法制备小鼠UC动物模型，试验共分为6组，其中饮用DSS的小鼠随机分为5组：粪肠球菌菌粉组、微囊化粪肠球菌组、布拉迪酵母菌菌粉组和微囊化布拉迪酵母菌组给予不同的药物治疗，药物皆以溶液形式灌胃给药，UC模型小鼠（DSS组）每天饮用与药物等体积的生理盐水；正常对照组灌服与药物等体积的生理盐水。观察UC小鼠的症状和组织学变化，ELSIA检测血清中细胞因子（IL-6、IL-10与TNF-α）的含量，Western blotting检测小鼠结肠黏膜紧密连接蛋白（Occludin和Claudin-1）表达量。结果显示，与模型组相比，治疗后各益生菌组小鼠血清中TNF-α和IL-6含量均显著降低（P<0.05），IL-10含量显著升高（P<0.05），微囊化益生菌组小鼠髓过氧化物酶（MPO）活性及结肠损伤组织学评分（粪肠球菌粉组除外）均显著下降（P<0.05），布拉迪酵母菌菌粉组与微囊化布拉迪酵母菌小鼠结肠黏膜内Occludin和Claudin-1表达量均显著升高（P<0.05）。粪肠球菌和布拉迪酵母菌可以缓解UC模型小鼠结肠炎症病变，且经过微囊化后的粪肠球菌和布拉迪酵母菌表现出更好的缓解小鼠急性溃疡性结肠炎病症效果，其机制与粪肠球菌和布拉迪酵母菌可以降低血清中IL-6与TNF-α含量，提高Occludin与Claudin-1的表达量密切相关。

关键词: 乳化凝胶化; 微囊化益生菌; 布拉迪酵母菌; 粪肠球菌; 结肠炎

Abstract: This study was aimed to observe the effects of microencapsulated Saccharomyces boulardii (S.boulardii) and Enterococcus faecium (E.faecalis) on ulcerative colitis (UC) and discuss its possible mechanism in mice.Mice model with UC was established by 3% dextran sulphate sodium (DSS),the test was divided into 6 groups,the model mice were randomized into 5 groups,mice were treated with unencapsulated-free E.faecalis,microencapsulated E.faecalis,unencapsulated-free S.boulardii and microencapsulated S.boulardii solution,respectively,and saline solution in DSS group by oral gavage.UC model mice (DSS group) were fed with the same volume of saline as the drug every day,while the normal control group was fed with the same volume of saline solution.The symptoms and histological changes of UC mice were observed,the pro-inflammatory cytokines including IL-6,IL-10 and TNF-α were measured by ELISA,and the expression of the Occludin and Claudin-1 were measured by Western blotting.The results showed that compared with DSS group,the contents of TNF-α and IL-6 were significantly reduced in probiotics groups (P<0.05),and the content of IL-10 was significantly increased (P<0.05).The myeloperoxidase activities and histoligical scores (except for unencapsulated-free E. faecalis group) were significantly decreased in microencapsulated pribiotics groups after treatment (P<0.05).The expression of Occludin and Claudin-1 were significantly increased inunencapsulated-free S.boulardii and microencapsulated S.boulardii groups(P<0.05).E.faecalis and S.boulardii could reduce the inflammatory lesions in the colon of UC mice,and the microencapsulated pribiotics had a better effect,moreover,the mechanisms of the improvement were possibly related to the decrease of the contents of IL-6 and TNF-α,and the increase of the expression of Occludin and Claudin-1.

Key words: emulsification gelation; microencapsulated probiotics; Saccharomyces boulardii; Enterococcus faecium; ulcerative colitis

中图分类号:

Q939.121

张琳, 綦文涛, 刘延国. 微囊化益生菌对小鼠急性溃疡性结肠炎的影响[J]. 中国畜牧兽医, 2020, 47(8): 2571-2578.

ZHANG Lin, QI Wentao, LIU Yanguo. Effects of Microencapsulated Probiotics on Ulcerative Colitis in Mice[J]. China Animal Husbandry and Veterinary Medicine, 2020, 47(8): 2571-2578.

参考文献

[1] SASAKI M,KLAPPROYH J M.The role of bacteria in the pathogenesis of ulcerative colitis[J].Journal of Signal Transduction,2012,2012:704953.
[2] LEE J S,CHA D S,PARK H J.Survival of freeze-dried Lactobacillus bulgaricus KFRI673 in chitosan-coated calcium alginate microparticles[J].Journal of Agricultural and Food Chemistry,2004,52(24):7300-7305.
[3] IYER C,PHILLIPS M,KAILASAPATHY K.Release studies of Lactobacillus casei strain shirota from chitosan-coated alginate-starch microcapsules in ex vivo porcine gastrointestinal contents[J].Letters in Applied Microbiology,2005,41(6):493-497.
[4] HOESLI C A,RAGHURAM K,KING R L,et al.Pancreatic cell immobilization in alginate beads produced by emulsion and internal gelation[J].Biotechnology and Bioengineering,2011,108(2):424-434.
[5] ALEXAKIS T,BOADI D K,QUONG D,et al.Microencapsulation of DNA within alginate microspheres and crosslinked chitosan membranes for in vivo application[J].Applied Biochemistry and Biotechnology,1995,50(1):93-106.
[6] GUAN H N,CHI D F,YU J,et al.Encapsulated ecdysone by internal gelation of alginate microspheres for controlling its release and photostability[J].Chemical Engineering Journal,2011,168(1):94-101.
[7] 李漫,袁杰利,刘党生.工业生产益生菌微囊化技术的研究进展[J].中国微生态学杂志,2014,26(3):354-357. LI M,YUAN J L,LIU D S.Microencapsulation technology in industrial production of probiotics:Research progress[J].Chinese Journal of Microecology,2014,26(3):354-357.(in Chinese)
[8] YANEZ-FERNANDEZ J,RAMOS-RAMIREZ E G,SALAZAR-MONTOYA J A. Rheological characterization of dispersions and emulsions used in the preparation of microcapsules obtained by interfacial polymerization containing Lactobacillus sp.[J].European Food Research and Technology,2008,226(5):957-966.
[9] 梁新晓,贠婷婷,田科雄,等.内源乳化凝胶化法制备海藻酸钙微胶珠的工艺优化[J].食品科学,2014,35(12):35-40. LIANG X X,YUN T T,TIAN K X,et al.Optimum preparation of calcium alginate microbeads by emulsification/internal gelation[J].Food Science,2014,35(12):35-40.(in Chinese)
[10] COOPER H S,MURTHY S N,SHAH R S,et al.Clinicopathologic study of dextran sulfate sodium experimental murine colitis[J].Laboratory Investigation,1993,69(2):238-249.
[11] FABIA R,AR'RAJAB A,JOHANSSON M L,et al.The effect of exogenous administration of Lactobacillus reuteri R2LC and oat fiber on acetic acid induced colitis in the rat[J].Scandinavian Journal of Gastroenterology,1993,28(2):155-162.
[12] DIELEMAN L A,PALMEN M J,AKOL H,et al.Chronic experimental colitis induced by dextran sulphatesodium (DSS) is characterized by Th1 and Th2 cytokines[J].Clinical and Experimental Immunology,1998,114(3):385-391.
[13] 李晓玲.美沙拉秦不同给药途径治疗活动期溃疡性结肠炎的疗效及对血清炎性因子的影响[J].河北医学,2016,22(10):1595-1597. LI X L.Effects of mesalazine by different administration routs on active ulcerative colitis and the influence on serum inflammatory factors[J].Hebei Medicine,2016,22(10):1595-1597.(in Chinese)
[14] DOTAN I,RACHMILEWITZ D.Probiotics in inflammatory bowel disease:Possible mechanisms of action[J].Current Opinion in Gastroenterology,2005,21(4):426-430.
[15] MADSEN K,CORNISH A,SOPER P,et al.Probiotic bacteria enhance murine and human intestinal epithelial barrier function[J].Gastroenterology,2001,121:580-591.
[16] 王旭霞,赵曙光,刘震雄,等.双歧杆菌对溃疡性结肠炎大鼠肠黏膜屏障功能及细胞因子的影响[J].山西医科大学学报,2011,42(8):621-623. WANG X X,ZHAO S G,LIU Z X,et al.Effects of Bifidobacterium on intestinal mucosal function and cytokines in rats with ulcerative colitis[J].Journal of Shanxi Medical University,2011,42(8):621-623.(in Chinese)
[17] THERESE M C,GREG L P,DAVID P F.Spotlight on VSL#3 probiotic mixture in chronic inflammatory bowel diseases[J].BioDrugs,2007,21(1):61-63.
[18] ROVEDATTI L,KUDO T,BIANCHERI P,et al.Different regulation of interleukin 17 and interferon gamma production in inflammatory bowel disease[J].Gut,2009,58(12):1629-1636.
[19] 韩桂华,遇常红,孙雪丹,等.青春双歧杆菌对溃疡性结肠炎患者血清相关因子的影响[J].中国老年学杂志,2016,36(2):888-889. HAN G H,YU C H,SUN X D,et al.Effect of Bifidobacterium adolescentis on serum related factors in patients with ulcerative colitis[J].Chinese Journal of Gerontology,2016,36(2):888-889.(in Chinese)
[20] MOHAMMAD R B,AZADEH M,PARVIN M,et al.Does cisapride,as a 5HT_4 receptor agonist,aggravate the severity of TNBS-induced colitis in rat[J].Gastroenterology Research and Practice,2012,2012:362536-362543.
[21] CANONICI A,SIRET C,PELLEGRINO E,et al.Saccharomyces boulardii improves intestinal cell restitution through activation of the a2b1 integrin collagen receptor[J].PLoS One,2011,6(3):e18427.
[22] 张晓明,王晓娟,徐洪雨,等.布拉氏酵母菌散对TNBS-大鼠溃疡性结肠炎IL-8、IFN-γ的表达影响[J].胃肠病学和肝病学杂志,2013,22(10):993-996. ZHANG X M,WANG X J,XU H Y,et al.Effect of Saccharomyces boulardii on the level of IL-8 and IFN-γ in TNBS-induced rat colitis[J].Chinese Journal of Gastroenterology and Hepatology,2013,22(10):993-996.(in Chinese)
[23] 马强,张方信,赵丽,等.乳杆菌对溃疡性结肠炎大鼠肠黏膜的影响[J].第四军医大学学报,2007,28(24):2242-2244. MA Q,ZHANG F X,ZHAO L,et al.Effect of Lactobacillus on colonic mucosa in rats with ulcerative colitis[J].Journal of the Fourth Military Medical University,2007,28(24):2242-2244.(in Chinese)
[24] OHIMA T,MIWA H,JOH T.Changes in the expression of claudins in active ulcerative colitis[J].Journal of Gastroenterology and Hepatology,2008,23(2):146-150.
[25] WEBER C R,NALLE S C,TRETIAKOVA M,et al.Claudin-1 and Claudin-2 expression is elevated in inflammatory bowel disease and may contribute to early neoplastic transformation[J].Laboratory Investigation,2008,88(10):1110-1120.
[26] MENNIGEN R,NOLTE K,RIJCKEN E,et al.Probiotic mixture VSL#3 protects the epithelial barrier by maintaining tight junction protein expression and preventing apoptosis in a murine model of colitis[J].AJP:Gastrointestinal and Liver Physiology,2009,296(5):1140-1149.
[27] SETH A,YAN F,POLK D B,et al.Probiotics ameliorate the hydrogen peroxide-induced epithelial barrierdisruption by a PKC- and MAP kinase-dependentmechanism[J].American Journal of Physiology:Gastrointestinal and Liver Physiology,2008,294(4):G1060-G1069.
[28] YEUNG C Y,CHIANG CHIAU J S,CHAN W T,et al.In vitro prevention of Salmonella lipopolysaccharide-induced damages in epithelial barrier function by various Lactobacillus strains[J].Gastroenterology Research and Practice,2013,2013:973209.
[29] 张中伟,秦环龙.乳酸菌对感染肠上皮细胞通透性及紧密连接蛋白表达的影响[J].肠外与肠内营养,2007,14(4):193-196. ZHANG Z W,QIN H L.Effect of Lactobacillus plantarunonlly regulation intestinal epithelial permeability and tight junction response to pathogenic bacteria[J].Parenteral & Enteral Nutrition,2007,14(4):193-196.(in Chinese)
[30] NANDA K N,BALAMURUGAN R,JAYAKANTHAN K,et al.Probiotic administration alters the gut flora and attenuates colitis in mice administered dextran sodium sulfate[J].Journal of Gastroenterology and Hepatology,2008,23(12):1834-1839.