Down-regulation of miR-186 contributes to podocytes apoptosis in membranous nephropathy
Down-regulation of miR-186 contributes to podocytes apoptosis in membranous nephropathy
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DOI：
10.1016/j.biopha.2015.07.021
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发表时间：
2015-10-01
影响因子：
7.5
通讯作者：
Lu, Guo-yuan
中科院分区：
文献类型：
--
作者：
Sha, Wen-gang;Shen, Lei;Lu, Guo-yuan
背景和目标：足细胞凋亡是膜性肾病发生发展的关键过程，miR-186与足细胞凋亡密切相关。本研究旨在探讨miR-186在膜性肾病（membranous nephropathy，MGN）患者足细胞凋亡中的作用及其机制。实时荧光定量PCR检测AngII诱导的肾组织和足细胞中miR-186的表达。Caspase-3活性检测足细胞凋亡。通过蛋白质印迹法定量TLR 4和P2 x 7蛋白表达。结果：MGN患者肾组织中miR-186的表达明显下调，而TLR 4和P2 x 7的表达明显上调。在体外实验中，TLR 4 siRNA增加了miR-186的表达，miR-186抑制剂增加了AngII暴露的足细胞中P2 x 7的mRNA和蛋白表达。此外，miR-186抑制剂可上调caspase-3的裂解水平。结论：miR-186模拟物可下调Ang Ⅱ诱导的足细胞凋亡的TUNEL阳性细胞数和caspase-3活性。(C)2015年由Elsevier Masson SAS出版。
Background and aim: Podocytes apoptosis is the key process in the development of membranous nephropathy and miR-186 is reported to be related with cell apoptosis. Here we investigated the expression of miR-186 in membranous nephropathy (MGN) patients and the mechanism underlying the podocytes apoptosis.Methods: Thirty patients with MGN and 30 healthy people were included in this study. The expression of miR-186 was detected in renal tissue and podocyte cells exposed to AngII by real-time PCR. Caspase-3 activity was used to evaluate podocytes apoptosis. TLR4 and P2 x 7 protein expression was quantified by western blotting. miR-186 inhibitor and miR-186 mimic were transfected into cells to investigate the mechanism underlying miR-186 in podocytes apoptosis.Results: In MGN patients, the level of miR-186 was significantly down-regulated as well as the protein expression of TLR4 and P2 x 7 was up-regulated in renal tissue. In vitro experiments, TLR4 siRNA increased the expression of miR-186 and miR-186 inhibitor elevated the mRNA and protein expression of P2 x 7 in podocytes exposed to AngII. In addition, the level of cleaved-caspase-3 was up-regulated by miR-186 inhibitor. The TUNEL-positive cells and caspase-3 activity of podocytes induced by AngII were down-regulated by miR-186 mimic.Conclusions: We revealed that TLR4 is involved in the regulation of miR-186 expression, and the antiapoptotic effect of miR-186 on podocytes is correlated with P2 x 7 regulation. (C) 2015 Published by Elsevier Masson SAS.