Angiotensin II receptor blocker valsartan ameliorates cardiac fibrosis partly by inhibiting miR-21 expression in diabetic nephropathy mice
Angiotensin II receptor blocker valsartan ameliorates cardiac fibrosis partly by inhibiting miR-21 expression in diabetic nephropathy mice
复制标题
血管紧张素 II 受体阻滞剂缬沙坦通过抑制糖尿病肾病小鼠的 miR-21 表达部分改善心脏纤维化
DOI：
10.1016/j.mce.2017.12.005
复制
发表时间：
2018-09-05
影响因子：
4.1
通讯作者：
Wang，Shaocheng
中科院分区：
文献类型：
--
作者：
Wang，Jinyang;Duan，Lijun;Wang，Shaocheng
糖尿病肾病（diabetic nephropathy，DN）是糖尿病的主要并发症之一。miR-21和MMP-9与纤维化疾病密切相关。血管紧张素II受体阻滞剂（ARB）具有心脏保护作用。然而，miR-21是否通过靶向MMP-9参与DN心肌纤维化的机制仍不清楚，ARB部分通过抑制miR-21的表达来减轻心肌纤维化。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。ISH显示，miR-21集中在细胞核附近的细胞质病灶中，主要定位于心脏成纤维细胞中，并且在DN心脏组织中的心肌细胞中处于相对低的水平。RT-PCR结果显示，糖尿病肾病患者心肌组织中miR-21表达明显增强，并伴有col-IV、FN、CVF、PVCA、LVMI、HWI和NT-pro-BNP的增加（p <0. 05）。生物信息学分析和荧光素酶报告基因分析表明，MMP-9是miR-21的有效靶点。此外，细胞转染实验表明，miR-21过表达直接降低MMP-9的表达。有趣的是，心肌组织中的miR-21水平与ACR呈正相关（r =-0.870，P = 0.003），而与SBP、HbA 1C和T-Cho不相关（p > 0.05）。更重要的是，ARB可显著降低心肌组织、心肌成纤维细胞和血清中miR-21的表达。总之，我们的研究结果表明，miR-21可能通过靶向MMP-9参与DN心肌纤维化的发病机制，miR-21可能是ARB治疗DN心肌纤维化的新的可能靶点。
Cardiac fibrosis with diabetic nephropathy (DN) is one of major diabetic complications. miR-21 and MMP-9 were closely associated with fibrosis diseases. Angiotensin II receptor blockers (ARB) have cardioprotective effects. However, it remains unclear whether miR-21 was involved in the mechanism of cardiac fibrosis with DN by target MMP-9 and ARB ameliorates cardiac fibrosis partly by inhibiting miR-21 expression. In this study, In Situ Hybridization(ISH), RT-PCR, cell transfection, western blotting and laser confocal telescope were used, respectively. ISH showed that miR-21, concentrated in cytoplasmic foci in the proximity of the nucleus, was mainly localized in cardiac fibroblasts and at relatively low levels in cardiomyocytes within cardiac tissue with DN. RT-PCR showed that miR-21 expression was significantly enhanced in cardiac tissue with DN, accompanied by the increase of col-IV, FN, CVF, PVCA, LVMI, HWI and NT-pro-BNP (p < 0.05). Bioinformatics analysis and Luciferase reporter gene assays showed that MMP-9 was a validated target of miR-21. Furthermore, cell transfection experiments showed that miR-21 overexpression directly decreased MMP-9 expression. Interestingly, miR-21 levels in cardiac tissue was positively correlated with ACR (r = −0.870, P = 0.003), whereas, uncorrelated with SBP, HbA1C and T-Cho (p > 0.05). More importantly, ARB can significantly decrease miR-21 expression in cardiac tissue, cardiac fibroblasts and serum. Overall, our results suggested that miR-21 may contribute to the pathogenesis of cardiac fibrosis with DN by target MMP-9, and that miR-21 may be a new possible therapeutic target for ARB in cardiac fibrosis with DN.