MicroRNA-1298调控GJA1影响下肢动脉硬化闭塞平滑肌细胞行为研究

项目名称： MicroRNA-1298调控GJA1影响下肢动脉硬化闭塞平滑肌细胞行为研究

项目编号： No.81200231

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学一处

项目作者： 王冕

作者单位： 中山大学

项目金额： 24万元

中文摘要： 动脉平滑肌细胞（ASMCs）增殖和迁移失控是导致下肢动脉硬化闭塞（ASO）的主要原因，microRNA在调控ASMCs行为变化中扮演重要角色，但是ASO中表达失调microRNA的功能仍不清楚。我们发现miR-1298在ASO中表达下调明显，升高miR-1298水平显著抑制ASMCs增殖，然而miR-1298对ASMCs迁移、凋亡等的影响以及其中的具体调控机制仍然未知，进一步分析发现可能与转录后水平调控GJA1表达有关。本研究拟采用microRNA转染、双荧光素酶报告基因、慢病毒表达、siRNA、原位杂交-免疫荧光双染等方法，旨在获得miR-1298通过GJA1调控ASMCs行为变化的可靠证据，评价和证实其在ASO发病中的作用。本项目将揭示ASO发病中miR-1298/GJA1调控ASMCs生物学行为变化的新机制，为确立miR-1298/GJA1作为干预ASO的新靶点提供更充分的科学依据。

中文关键词： 微小RNA；动脉平滑肌细胞；缝隙连接蛋白；下肢动脉硬化闭塞；DNA甲基化

英文摘要： Excessive artery smooth muscle cell (ASMCs) proliferation and migration play pivotal steps in the genesis and process of arteriosclerosis obliterans (ASO). MicroRNA is recognized as an important governor of ASMCs behavior by regulating target gene expression at post-transcriptional level. Recently, we reported the microRNA expression profile in ASO samples using microRNA real-time PCR array screening. Among the most dysregulated microRNAs, miR-1298 was found downregulated in ASO media dramatically. Further introducing miR-1298 into ASMCs using miR-1298 mimic transfection significantly attenuated mitogens triggered ASMCs proliferation. It indicates that loss of miR-1298 expression might be related with ASO. However, the exact role and underlying mechanisms of miR-1298 regulating ASMCs behavior in ASO remains unknown. Other than microRNAs, disordered intercellular communication between vascular cells is also an important initiator to promote ASMCs proliferation and migration. Gap junction is the predominant intercellular communication manner between ASMCs, mainly consists of Gap Junction protein Alpha-1 (GJA1). GJA1 upregulation was found in stenotic artery disease, and deletion of GJA1 was proved to be efficient in preventing artery restenosis in animal models. Our data also showed increased GJA1 protein express

英文关键词： microRNA；artery smooth muscle cell；gap junction protein alpha 1；arteriosclerosis obliterans；DNA methylation