MicroRNA-217 suppresses homocysteine-induced proliferation and migration of vascular smooth muscle cells via N-methyl-D-aspartic acid receptor inhibition
MicroRNA-217 suppresses homocysteine-induced proliferation and migration of vascular smooth muscle cells via N-methyl-D-aspartic acid receptor inhibition
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DOI：
10.1111/1440-1681.12611
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发表时间：
2016-10-01
影响因子：
2.9
通讯作者：
Wang, Shanling
中科院分区：
文献类型：
--
作者：
Duan, Hongyan;Li, Yongqiang;Wang, Shanling
高同型半胱氨酸已成为动脉粥样硬化的重要危险因素，并可刺激血管平滑肌细胞（VSMCs）的增殖和迁移。N-甲基-D-天冬氨酸受体（NMDAR）是同型半胱氨酸的受体，介导同型半胱氨酸对血管平滑肌细胞的作用。生物信息学分析显示NMDAR是microRNA-217（miR-217）的潜在靶点，其在癌症肿瘤发生和颈动脉斑块进展中发挥多种功能。在这项研究中，我们试图研究miR-217在同型半胱氨酸暴露下VSMCs表型转变中的作用，并阐明其对动脉粥样硬化斑块形成的影响。不同浓度的同型半胱氨酸（0- 8 × 10（-4）mol/L）作用24 h后，HA-VSMCs和大鼠主动脉VSMCs中miR-217的表达无明显变化。有趣的是，同型半胱氨酸以剂量依赖性方式降低NMDAR mRNA和蛋白的表达。转染miR-217模拟物显著抑制同型半胱氨酸处理的VSMCs增殖和迁移，而转染miR-217抑制剂促进VSMCs迁移。此外，miR-217模拟物下调而miR-217抑制剂上调NMDAR蛋白表达，但不上调NMDAR mRNA表达。通过荧光素酶报告基因分析，我们发现miR-217可以直接与NMDAR的3-UTR结合。miR-217模拟转染也释放了由同型半胱氨酸诱导的cAMP-反应元件结合蛋白（CREB）-PGC-1信号转导的抑制。此外，通过AdPGC-1感染恢复PGC-1表达通过NADPH氧化酶（NOX 1）的降解和活性氧（ROS）的减少显著抑制VSMCs增殖。总之，我们的研究确定了miR-217在调节VSMCs增殖和迁移中的作用，这可能成为动脉粥样硬化治疗的靶点。
Hyperhomocysteine has become a critical risk for atherosclerosis and can stimulate proliferation and migration of vascular smooth muscle cells (VSMCs). N-methyl-D-aspartic acid receptor (NMDAR) is a receptor of homocysteine and mediates the effects of homocysteine on VSMCs. Bioinformatics analysis has shown NMDAR is a potential target of microRNA-217 (miR-217), which exerts multiple functions in cancer tumorigenesis and carotid plaque progression. In this study, we sought to investigate the role of miR-217 in VSMCs phenotype transition under homocysteine exposure and elucidate its effect on atherosclerotic plaque formation. After treating with several doses of homocysteine (0-8x10(-4)mol/L) for 24hours, the expression of miR-217 in HA-VSMCs and rat aortic VSMCs was not altered. Intriguingly, the expression of NMDAR mRNA and protein was reduced by homocysteine in a dose-dependent manner. Transfection of miR-217 mimic significantly inhibited the proliferation and migration of VSMCs with homocysteine treatment, while transfection of miR-217 inhibitor promoted VSMCs migration. Moreover, miR-217 mimic down-regulated while miR-217 inhibitor up-regulated NMDAR protein expression but not NMDAR mRNA expression. Through luciferase reporter assay, we showed that miR-217 could directly bind to the 3-UTR of NMDAR. MiR-217 mimic transfection also released the inhibition of cAMP-response element-binding protein (CREB)-PGC-1 signalling induced by homocysteine. Additionally, restoration of PGC-1 expression via AdPGC-1 infection markedly suppressed VSMCs proliferation through the degradation of NADPH oxidase (NOX1) and reduction of reactive oxygen species (ROS). Collectively, our study identified the role of miR-217 in regulating VSMCs proliferation and migration, which might serve as a target for atherosclerosis therapy.