RIP2对大肠杆菌的抗菌作用研究  

Expression of RIP2 enhances clearance of intracellular Escherichia coli in SW480 cells

作　　者：席琼[1] 胡巢凤[1] 张芸[1] 陆大祥[1] 蒋宇[1] 

机构地区：[1]暨南大学医学院病理生理学教研室国家中医药管理局三级科研实验室,广东广州510632

出　　处：《中国病理生理杂志》2011年第5期951-955,共5页Chinese Journal of Pathophysiology

基　　金：广东省自然科学基金资助项目(No.06025159);广东省教育厅自然科学研究项目[No.粤财教(2005)126];暨南大学"211工程"三期预研项目;暨南大学重点实验室基金资助项目

摘　　要：目的:探讨受体相互作用蛋白2(RIP2)对人肠细胞清除大肠杆菌的作用。方法:使用阳离子聚合物JetPeiTM介导人RIP2基因(pEGFP-C2-PIP2)转染人结肠癌SW480细胞株,以转染空质粒及未转染的SW480细胞株作为对照,应用RT-PCR检测外源性RIP2 mRNA水平的变化、Western blotting法检测细胞RIP2蛋白的表达;并利用大肠杆菌ATCC25922感染转染和未转染的SW480细胞24 h,用平板培养菌落计数活菌数。应用p38MAPK通路抑制剂SB203580作用于3组细胞,计数活菌数。结果:与对照组相比,转染RIP2组其mRNA和蛋白表达水平均有明显升高(P<0.01,P<0.05),表明RIP2表达质粒成功转染SW480细胞。转染RIP2细胞能清除胞内大肠杆菌;而阻断p38 MAPK信号通路后,RIP2清除胞内大肠杆菌的作用被阻断。结论:RIP2转染细胞可有效清除胞内大肠杆菌,这种胞内细菌的清除作用可能与p38 MAPK信号通路有关。这些结果提示RIP2在细菌感染性疾病治疗中具有广阔的临床应用前景。AIM： To investigate the clearance effect and mechanism of high expression of receptor-interacting protein 2（RIP2） against Escherichia coli（E.coli） in intestinal cells.METHODS： RIP2 gene（pEGFP-C2-PIP2） was transfected into cell line SW480 by the method of JetPeiTM.Western blotting and RT-PCR analysis were then used to detect RIP2 protein and mRNA expression,respectively.The transfected and non-transfected cells were incubated with E.coli at 37 ℃.At 24 h after initial incubation,the viable intracellular bacteria were quantified by plating appropriate dilution on LB agar plates.The transfected and non-transfected cells were treated with SB203580,an inhibitor of p38 MAPK pathway,and the changes of intracellular bacteria were also quantified.RESULTS： Compared with the cells transfected with pEGFP-C2 and non-transfected cells,the expression of RIP2 at mRNA and protein levels was significantly enhanced in SW480 cells transfected with recombinant plasmid pEGFP-C2-PIP2（P0.01 and P0.05,respectively）.The cells transfected with pEGFP-C2-PIP2 had the ability to clear invading E.coli,and this antibacterial effect was inhibited by a p38 MAPK inhibitor SB203580.CONCLUSION： RIP2 effectively eliminates the invading E.coli,and p38 MAPK pathway plays an important role in the clearance effect of RIP2,indicating that RIP2 may be a potential molecular therapeutic target for infectious disease.

关 键 词：受体相互作用蛋白2 基因转染 抗菌作用 

分 类 号：R392.11[医药卫生—免疫学]