线粒体功能障碍参与糖尿病肾脏疾病足细胞损伤的初步研究

Role of mitochondrial dysfunction in podocyte injury in diabetic kidney disease

目的探讨糖尿病肾脏疾病足细胞是否存在线粒体功能障碍。方法分别从2型糖尿病db/db小鼠及其同窝对照db/m小鼠肾脏中分离足细胞,提取线粒体检测线粒体复合物Ⅰ和Ⅲ功能,提取RNA行实时定量PCR检测足细胞线粒体代谢相关基因的表达;采用免疫组织化学检查检测足细胞标记蛋白WT1的表达。结果 db/db小鼠足细胞线粒体复合物Ⅰ和复合物Ⅲ的活性、线粒体DNA拷贝数均较db/m小鼠低(P均<0.05),db/db小鼠足细胞与线粒体代谢相关的基因SIRT1、PGC-1α、TFAM、NRF1、NRF2以及足细胞标记蛋白nephrin mRNA的表达均较db/m小鼠低(P均<0.05),db/db小鼠足细胞数量较db/m小鼠少(P<0.05)。结论糖尿病小鼠足细胞存在线粒体功能障碍。

Objective To investigate the presence of mitochondrial dysfunction in the podocytes of mouse models with diabetic kidney disease. Methods The podocytes were isolated from the kidney from db/db mice with type 2 diabetes mellitus and db/m control mice. The mitochondria was extracted for detection of the function of mitochondrial complex I and III. RNA extraction was performed. Real-time quantitative PCR was performed to detect the genes related to podocyte mitochondrial metabolism. The expression of podocyte-labeled protein WT1 was measured by immunohistochemical staining. Results In the db/db mice, the activity of podocyte mitochondrial complex I and III , and the copy number of mitochondrial DNA were significantly lower compared with those in the db/m mice （all P 〈 0. 05 ）. The expression levels of SIRT1, PGC-1α, TFAM, NRF1, NRF2 and podocyte-labeled protein nephrin mRNA in the db/db mice were significantly down-regula-ted compared with those in db/m mice （all P 〈0. 05）. The quantity of podocytes in the db/db mice was con-siderably less than that in the db/m mice （ P 〈 0. 05 ）. Conclusion Mitochondrial dysfunction occurs in the podocytes of diabetes mellitus mice.