益生菌对炎性肠病小鼠肠屏障功能的影响  

Effect of Probiotics on Intestinal Barrier Function in Mice with Inflammatory Bowel Disease

作　　者：陈红旗[1] 杨俊[1] 周玉坤[1] 张旻 杭晓敏 秦环龙[1] 

机构地区：[1]上海交通大学附属第六人民医院普外科,上海200233 [2]上海交大昂立医药研究院,上海200233

出　　处：《中国普外基础与临床杂志》2011年第12期1242-1247,共6页Chinese Journal of Bases and Clinics In General Surgery

基　　金：黎介寿院士肠道屏障研究专项基金资助项目(项目编号:LJS-2009001)~~

摘　　要：目的研究益生菌植物乳杆菌(LP)对炎症状态下肠屏障的修复机理。方法以白介素-10基因敲除(IL-10-/-)小鼠为炎性肠病模型,采用简单随机法(随机表)将空白对照(WT)小鼠和IL-10-/-小鼠分成4组:WT组、WT+LP组、IL-10-/-组和IL-10-/-+LP组,每组10只。分别灌饲Ringer液或LP溶液,持续4周。每周记录小鼠肠炎的临床表现,实验结束后收集小鼠结肠组织,进行病理评分和形态观察,ELISA法检测结肠组织促炎细胞因子肿瘤坏死因子-α(TNF-α)和干扰素-γ(IFN-γ)浓度,利用Ussing Chamber技术对小鼠结肠通透性及电生理变化进行分析,并用Western blot和免疫荧光分析法对结肠上皮细胞紧密连接蛋白进行定量和定位研究。结果与WT组相比,IL-10-/-组小鼠腹泻、直肠脱垂和体重丧失较明显(P<0.01);TNF-α和IFN-γ浓度明显升高(P<0.01);病理表现为大量炎性细胞浸润,偶见透壁性溃疡和隐窝脓肿形成;超微电镜提示紧密连接结构破坏;紧密连接蛋白(ZO-1、occludin、claudin-1)表达明显降低(P<0.01)且伴有异常分布;结肠上皮细胞甘露醇通透性升高(P<0.001),跨膜电阻值降低(P<0.001)。LP干预4周后,与IL-10-/-组相比,LP能显著改善肠道炎症的临床和病理表现,阻止紧密连接超微结构破坏,促进紧密连接蛋白的表达(P<0.01),降低结肠上皮细胞甘露醇通透性(P<0.001),升高跨膜电阻值(P<0.001),降低了TNF-α和IFN-γ的浓度(P<0.01),减轻了肠道炎症。结论 LP能通过促进肠上皮细胞间紧密连接蛋白的表达修复受损肠屏障功能,从而减轻肠道炎症。Objective To investigate the effect of Lactobacillus plantarum（LP） on the intestinal barrier function under inflammation. Methods Interleukin-10 knockout（IL-10-/-） mice were used as the model of inflammatory bowel disease.IL-10-/-and wild type（WT） mice received the LP or Ringer solutions for 4 weeks.Colitis was assessed by histological score and clinical manifestation was observed.The gut paracellular permeability was measured by Ussing chamber.The concentrations of tumor necrosis factor-α（TNF-α） and interferon-γ（IFN-γ） were detected by the ELISA method. The expressions and distributions of tight junction proteins were determined by Western blot and immunofluorescence,respectively. Results Compared with the WT group,the diarrhea,rectal prolapse,and weight loss were obvious（P〈0.01）,the concentrations of TNF-α and IFN-γ significantly increased（P〈0.01）,the infiltration of numerous inflammatory cells,even transmural ulcers,and crypt abscess were observed,the ultrastructure of tight junction was damaged,the mannitol permeability significantly increased（P〈0.001） and transepithelial resistance（TER） significantly decreased（P〈0.001）,and the expressions of tight junction proteins（ZO-1,occludin,and claudin-1） significantly decreased（P〈0.01） in the IL-10-/-group.Compared with the IL-10-/-group,the clinical and pathological manifestations of colitis significantly improved（P〈0.01）,the ultrastructural damage of tight junction was prevented,the mannitol permeability significantly decreased（P〈0.001） and the TER significantly increased（P〈0.001）,the concentrations of TNF-α and IFN-γ significantly decreased（P〈0.01）,and the expressions of tight junction proteins（ZO-1,occludin,and claudin-1） significantly increased（P〈0.01） in the IL-10-/-＋LP group. Conclusion Treatment with LP ameliorates colonic epithelial barrier dysfunction by promoting the expressions of tight junctional proteins in IL-10-/-mice.

关 键 词：益生菌 植物乳杆菌 肠屏障 紧密连接蛋白 

分 类 号：R574.62[医药卫生—消化系统]