Downregulation of the miR-30 family microRNAs contributes to endoplasmic reticulum stress in cardiac muscle and vascular smooth muscle cells
Downregulation of the miR-30 family microRNAs contributes to endoplasmic reticulum stress in cardiac muscle and vascular smooth muscle cells
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DOI：
10.1016/j.ijcard.2014.02.007
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发表时间：
2014-04-15
影响因子：
3.5
通讯作者：
Zhang, Jianjun
中科院分区：
文献类型：
--
作者：
Chen, Ming;Ma, Guiling;Zhang, Jianjun
背景资料：内质网（ER）应激是一种常见的亚细胞应激反应，其核心是葡萄糖调节蛋白78（GRP 78）的表达增加。然而，GRP 78上调的机制仍然知之甚少。方法：采用H2 O2诱导新生大鼠心室肌细胞（NRVC）和大鼠主动脉平滑肌细胞（RAVSMCs）的内质网应激模型。分子生物学技术用于定量蛋白质和mRNA水平。采用荧光素酶报告基因分析法研究miRNA的靶向性。结果：H2 O2诱导心血管NRVC和RAVSMCs内质网应激后，miR-30家族的miR-30 a、B、c、d和e均下调，同时沿着GRP 78、切割的ATF 6、CHOP和切割的caspase-12上调。证实GRP 78是miR-30的靶基因。通过antimiR-30人工敲低miR-30引发了表型ER应激，其中GRP 78/ATF 6/CHOP/caspase-12显著上调和细胞死亡，而miR-30替代减轻了ER应激。通过siRNA敲低CHOP可调控miR-30家族的所有成员，而通过其诱骗物螯合C/EBP转录因子可下调miR-30 miRNA.Conclusions：总体而言，miR-30家族miRNAs的下调有助于ER应激和心血管系统中GRP 78的相关上调。miR-30的参与在ER应激信号通路中产生正反馈环。MiR-30替代可能是缓解与ER应激相关的疾病的可行方法。(C)2014爱思唯尔爱尔兰有限公司版权所有。
Background: Endoplasmic reticulum (ER) stress is a common subcellular response to stresses and central to ER stress is increased expression of glucose-regulated protein 78 (GRP78). However, the mechanisms for GRP78 upregulation remained poorly understood. Our study goal was to shed light on this issue.Methods: H2O2 was used to create cellular models of ER stress in neonatal rat ventricular cells (NRVCs) and rat aorta vascular smooth muscle cells (RAVSMCs). Molecular Biology techniques were used to quantify protein and mRNA levels. Luciferase reporter gene assay was employed to investigate miRNA targeting. MTT assay and ELISA were used to detect cell death.Results: MiRNAs belonging to the miR-30 family including miR-30a, b, c, d and e were all downregulated in ER stress induced by H2O2 in cardiovascular cells NRVCs and RAVSMCs, along with the upregulation of GRP78, cleaved ATF6, CHOP, and cleaved caspase-12. GRP78 was confirmed to be a target gene for miR-30. Artificial knockdown of miR-30 by antimiR-30 triggered the phenotypic ER stress with significant GRP78/ATF6/CHOP/caspase-12 upregulations and cell death, while miR-30 replacement mitigated ER stress. Knockdown of CHOP by siRNA regulated all members of the miR-30 family whereas sequestration of C/EBP transcription factor by its decoy downregulated miR-30 miRNAs.Conclusions: Collectively, downregulation of the miR-30 family miRNAs contributes to the ER stress and the associated upregulation of GRP78 in the cardiovascular system. The participation of miR-30 creates a positive feedback loop in the ER stress signaling pathway. MiR-30 replacement may be a viable approach for alleviating disorders associated with ER stress. (C) 2014 Elsevier Ireland Ltd. All rights reserved.