微小RNA-26a通过靶向瞬时受体电位阳离子通道蛋白3抑制动脉粥样硬化进展。
miR-26a inhibits atherosclerosis progression by targeting TRPC3.
作者信息
Feng Min, Xu Daqian, Wang Lirui
机构信息
Department of Intensive Care Unit, The First Affiliated Hospital of Zhengzhou University, No. 1 of Jian She East Road, Zhengzhou, 450052 China.
出版信息
Cell Biosci. 2018 Jan 19;8:4. doi: 10.1186/s13578-018-0203-9. eCollection 2018.
BACKGROUND
Atherosclerosis, a chronic multi-factorial vascular disease, has become a predominant cause of a variety of cardiovascular disorders. miR-26a was previously reported to be involved in atherosclerosis progression. However, the underlying mechanism of miR-26a in atherosclerosis remains to be further explained.
METHODS
High-fat diet (HFD)-fed apolipoprotein E (apoE) mice and oxidized low-density lipoprotein (ox-LDL)-stimulated human aortic endothelial cells (HAECs) were established as in vivo and in vitro models of atherosclerosis. RT-qPCR and western blot analysis were performed to measure the expression of miR-26a and transient receptor potential canonical 3 (TRPC3), respectively. Binding between miR-26a and TRPC3 was predicted with bioinformatics software and verified using a dual luciferase reporter assay. The effects of miR-26a on the lipid accumulation, atherosclerotic lesion, and inflammatory response in HFD-fed apoE mice were investigated by a colorimetric enzymatic assay system, hematoxylin-eosin and oil-Red-O staining, and ELISA, respectively. Additionally, the effects of miR-26a or combined with TRPC3 on cell viability, apoptosis and the nuclear factor-kappa B (NF-κB) pathway in ox-LDL-stimulated HAECs were evaluated by MTT assay, TUNEL assay, and western blot, respectively.
RESULTS
miR-26a was downregulated in HFD-fed apoE mice and ox-LDL-stimulated HAECs. miR-26a overexpression inhibited the pathogenesis of atherosclerosis by attenuating hyperlipidemia, atherosclerotic lesion and suppressing inflammatory response in HFD-fed apoE mice. Moreover, miR-26a overexpression suppressed inflammatory response and the NF-κB pathway, promoted cell viability and inhibited apoptosis in ox-LDL-stimulated HAECs. Additionally, TRPC3 was demonstrated to be a direct target of miR-26a. Enforced expression of TRPC3 reversed the effects of miR-26a on cell viability, apoptosis, and the NF-κB pathway in ox-LDL-treated HAECs.
CONCLUSIONS
miR-26a alleviated the development of atherosclerosis by regulating TRPC3, providing a potential target for atherosclerosis treatment.
背景
动脉粥样硬化是一种慢性多因素血管疾病，已成为多种心血管疾病的主要病因。先前有报道称miR-26a参与动脉粥样硬化的进展。然而，miR-26a在动脉粥样硬化中的潜在机制仍有待进一步阐明。
方法
将高脂饮食（HFD）喂养的载脂蛋白E（apoE）小鼠和氧化型低密度脂蛋白（ox-LDL）刺激的人主动脉内皮细胞（HAECs）分别作为动脉粥样硬化的体内和体外模型。分别采用RT-qPCR和蛋白质印迹分析来检测miR-26a和瞬时受体电位阳离子通道亚家族C成员3（TRPC3）的表达。使用生物信息学软件预测miR-26a与TRPC3之间的结合，并通过双荧光素酶报告基因检测进行验证。分别通过比色酶法检测系统、苏木精-伊红染色和油红O染色以及ELISA，研究miR-26a对HFD喂养的apoE小鼠脂质蓄积、动脉粥样硬化病变和炎症反应的影响。此外，分别通过MTT法、TUNEL法和蛋白质印迹，评估miR-26a或与TRPC3联合对ox-LDL刺激的HAECs细胞活力、凋亡和核因子κB（NF-κB）通路的影响。
结果
在HFD喂养的apoE小鼠和ox-LDL刺激的HAECs中，miR-26a表达下调。miR-26a过表达通过减轻HFD喂养的apoE小鼠的高脂血症、动脉粥样硬化病变并抑制炎症反应，抑制动脉粥样硬化的发病机制。此外，miR-26a过表达抑制了ox-LDL刺激的HAECs中的炎症反应和NF-κB通路，促进细胞活力并抑制细胞凋亡。此外，TRPC3被证明是miR-26a的直接靶点。TRPC3的过表达逆转了miR-26a对ox-LDL处理的HAECs细胞活力、凋亡和NF-κB通路的影响。
结论
miR-26a通过调节TRPC3减轻动脉粥样硬化的发展，为动脉粥样硬化治疗提供了一个潜在靶点。
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