Downregulation of miR-30c promotes renal fibrosis by target CTGF in diabetic nephropathy
Downregulation of miR-30c promotes renal fibrosis by target CTGF in diabetic nephropathy
复制标题
糖尿病肾病中miR-30c的下调通过靶标CTGF促进肾纤维化
DOI：
10.1016/j.jdiacomp.2015.12.011
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发表时间：
2016-04-01
影响因子：
3
通讯作者：
Yang, Jinkui
中科院分区：
文献类型：
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作者：
Wang, Jinyang;Duan, Lijun;Yang, Jinkui
microRNA（miRs）在许多病理过程的发生和发展中起重要作用。然而，miR-30 c在糖尿病肾病（DN）中的作用仍不清楚。本研究旨在探讨miR-30 c是否通过抑制靶向CTGF的表达而参与DN的肾纤维化机制。本研究分别采用原位杂交、RT-PCR、细胞转染、蛋白质印迹和激光共聚焦显微镜等技术。原位杂交结果显示，miR-30 c主要分布于皮质内的肾小球和肾小管上皮细胞，集中于细胞核附近的细胞质中。RT-PCR结果显示，DN组miR-30 c表达明显降低（p < 0.05），与ISH结果一致。荧光素酶报告基因分析表明CTGF是miR-30 c的有效靶点。此外，miR-30 c过表达直接降低CTGF mRNA和蛋白。相反，miR-30 c抑制剂增强CTGF表达。miR-30 c表达与ACR呈负相关（r =-0.870，P = 0.003），与Ccr呈正相关（r = 0.8230，P = 0.01），而与KW/BW、SBP、HbA 1C、HOMR-IR和T-Cho无相关性（p > 0.05）。更重要的是，miR-30 c模拟物显著降低col-IV、FN、GSI、GBM、GA、MRA/CLA和ACR（p < 0.05），相反，轻微但显著增加Ccr（p < 0.05）。总之，我们的研究结果表明，miR-30 c的缺失可能通过抑制靶CTGF表达而参与DN的发病机制;补充miR-30 c可能通过减少DN中的肾纤维化而改善肾脏结构和功能。(C)2016 Elsevier Inc. All rights reserved.
MicroRNAs (miRs) play important roles in initiation and progression of many pathologic processes. However, the role of miR-30c in diabetic nephropathy (DN) remains unclear. This study was to determine whether miR-30c was involved in the mechanism of renal fibrosis by inhibiting target CTGF expression in DN. In this study, In Situ Hybridization(ISH), RT-PCR, cell transfection, western blotting and laser confocal telescope were used, respectively. ISH showed that miR-30c, concentrated in cytoplasmic foci in the proximity of the nucleus, was mainly localized in glomerular and renal tubular epithelial cells within the cortex. RT-PCR showed that miR-30c expression was significantly decreased in DN (p < 0.05), consistent with of the results of ISH. Luciferase reporter gene assays showed that CTGF was a validated target of miR-30c. Furthermore, miR-30c overexpression directly decreased CTGF mRNA and protein. Conversely, miR-30c inhibitor enhanced CTGF expression. Interestingly, miR-30c expression was negatively correlated with ACR (r = -0.870, P = 0.003) and positively correlated with Ccr (r = 0.8230, P = 0.01), whereas it was uncorrelated with KW/BW, SBP, HbA1C, HOMR-IR and T-Cho (p > 0.05). More importantly, miR-30c mimics significantly decreased col-IV, FN, GSI, GBM, GA, MRA/CLA and ACR (p < 0.05) and, in contrast, slightly but significantly increased Ccr (p < 0.05). In conclusion, our results suggested that loss of miR-30c may contribute to the pathogenesis of DN by inhibiting target CTGF expression; replenishing miR-30c may ameliorate renal structure and function by reducing renal fibrosis in DN. (C) 2016 Elsevier Inc. All rights reserved.