MiR-135a与糖尿病肾病肾脏纤维化的相关性研究  

Investigate the relationship between miR-135a and renal fibrosis in diabetic nephropathy

作　　者：刘思逸 何凤[1] 陈钦开[1] 魏昕[1] 张莉[1] 杨玉娟 

机构地区：[1]南昌大学第一附属医院肾内科,南昌330006

出　　处：《江西医药》2016年第7期624-629,共6页Jiangxi Medical Journal

基　　金：江西省科技厅自然科学基金资助课题,编号:2014BAB205015

摘　　要：目的探讨miR-135a与糖尿病肾病肾脏纤维化关系及其具体作用机制。方法通过RT-PCR技术测定糖尿病肾病患者血清及肾脏组织中miRNA-135a(miR-135a)的含量。采用荧光素酶报告基因实验验证miRNA-135a的靶基因。构建miR-135a过表达质粒,上调肾小球系膜细胞及肾小管上皮细胞中miR-135a的表达,通过蛋白印记及免疫荧光法检测与肾脏纤维化相关蛋白的表达,运用荧光素报告基因及免疫印记的方法寻找、确定miR-135a。结果从糖尿病肾病患者以及db/db小鼠的血清和肾组织的miR-135a水平来看,miR-135a表达显著上调。我们证实了瞬时受体电位阳离子通道,亚家族C,成员1(Transient receptor potential channel,subfamily C,member 1,TRPC1)是miR-135a的靶基因。TRPC1的过度表达能够逆转miR-135a对促进肾小球系膜细胞增殖以及增加胞外基质蛋白合成的病理效应。在糖尿病患者肾组织中miR-135a的减少使得TRPC1回到正常水平,并减少了纤维连接蛋白和胶原蛋白I在人体内的合成。抑制TRPC1水平可能是miR-135a在糖尿病肾损伤中促肾纤维化的一种机制。结论 miR-135a可通过靶向调节TRPC1参与糖尿病肾病肾脏纤维化的发生,这些发现表明miR-135a在肾脏纤维化中有重要作用,抑制miR-135a的表达可能成为治疗糖尿病肾病的一种有效方法。Objective To investigate the relationship between miR-135 a and renal fibrosis in diabetic nephropathy and the mechanism. Methods The abundance of miRNA-135a（miR-135a） was measured by real-time quantitative PCR in the serum and kidney tissues of patients with diabetic nephropathy. The luciferase assay combined with mutation and immunoblotting was used to screen and verify the bioinformatically predicted miRNAs. Results MiR-135 a was markedly upregulated in serum and renal tissue from patients with diabetic nephropathy,as well from db/dbmice. Furthermore,we identified transient receptor potential cation channel,subfamily C,member 1（TRPC1） as a target of miR-135 a during renal injury. We demonstrated that overexpression of TRPC1 was able to reverse the pathological effects of miR-135 a on promoting proliferation of mesangial cells and increasing synthesis of extracellular matrix proteins. Importantly,knockdown of miR-135 a in diabetic kidneys restored levels of TRPC1 and reduced synthesis of fibronectin and collagen I in vivo. Suppressing TRPC1 levels may be a mechanism whereby miR-135 a promotes renal fibrosis in diabetic kidney injury. Conclusion These findings suggest an important role for miR-135 a in renal fibrosis and inhibition of miR-135 a might be an effective therapy for diabetic nephropathy.

关 键 词：糖尿病肾病 纤维化 microRNA-135a TRPC1 

分 类 号：R587.1[医药卫生—内分泌]