MicroRNA-429靶向抑制occludin及对糖尿病小鼠肠上皮屏障功能的影响  

Effect of targeted inhibition of occludin by microRNA-429 on intestinal epithelial barrier function in diabetic mouse

作　　者：卢锡基 于涛[1] 夏忠胜[1] 单体栋[1] 欧阳慧[1] 许稷豪 陈其奎[1] 

机构地区：[1]中山大学孙逸仙纪念医院消化内科,广东广州510120

出　　处：《中国病理生理杂志》2016年第4期696-700,共5页Chinese Journal of Pathophysiology

基　　金：国家自然科学基金资助项目(No.81370475)

摘　　要：目的:探讨microRNA-429(miR-429)在体内对糖尿病(DM)小鼠肠上皮细胞(IECs)内闭合蛋白(occludin,Ocln)表达及肠上皮屏障功能的影响。方法:构建糖尿病小鼠模型(腹腔内注射链脲佐菌素);通过尾静脉注射antagomiRNA-429抑制DM小鼠IECs内miR-429的表达;real-time PCR检测miR-429和Ocln mRNA的表达变化;Western blotting及免疫组化检测Ocln蛋白表达变化;气相色谱法检测小鼠尿液乳果糖/甘露醇比值;显色基质鲎试剂法检测小鼠血浆LPS浓度。结果:尾静脉注射antagomiRNA-429能显著抑制DM小鼠IECs内miR-429的表达,注射后6 d恢复至正常小鼠IECs表达水平。抑制DM小鼠IECs内miR-429表达后,Ocln表达显著提高(P<0.05),小鼠尿液乳果糖/甘露醇比值及血浆LPS水平均明显下降(P<0.05)。结论:AntagomiRNA-429能有效抑制DM小鼠IECs内miR-429的表达,进而使小鼠IECs内Ocln的表达升高,从而使肠上皮屏障功能部分恢复。AIM： To investigate the effect of microRNA-429（ miR-429） on the expression of occludin（ Ocln）in intestinal epithelial cells（ IECs） and intestinal epithelial barrier function in diabetic mice. METHODS： Diabetes mellitus mouse model was induced by intraperitoneal injection of streptozocin. The expression of miR-429 in IECs of diabetic mice was inhibited by antagomiRNA-429 injected via tail vein. The expression of miR-429 and mRNA expression of Ocln were detected by real-time PCR. The protein expression of Ocln was determined by Western blotting and immunohistochemistry. The urinary lactulose / mannitol ratio was measured by gas chromatography. The plasma LPS concentrations in the mice were measured by chromogenic end-point TAL kit. RESULTS： The results of real-time PCR confirmed that the expression of miR-429 in IECs of diabetic mice was remarkably inhibited by tail-vein administration of antagomiRNA-429,and resumed to similar level of normal mice on the 6th day after the administration. After suppressing the level of miR-429,the expression of Ocln in IECs of diabetic mice increased significantly（ P 0. 05）,while the urinary lactulose / mannitol ratio and the plasma LPS concentration decreased obviously（ P 0. 05）. CONCLUSION： AntagomiRNA-429 effectively suppresses miR-429 expression in IECs of diabetic mice,and then enhances the expression of Ocln and partially resumes the intestinal epithelial barrier function.

关 键 词：糖尿病 MicroRNA-429 闭合蛋白 肠上皮屏障功能 

分 类 号：R363.2[医药卫生—病理学] R587.1[医药卫生—基础医学]