GSK3β调控内质网应激介导的血管平滑肌细胞增殖异常的机制

项目名称： GSK3β调控内质网应激介导的血管平滑肌细胞增殖异常的机制

项目编号： No.81201344

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学四处

项目作者： 楚元奎

作者单位： 宁夏医科大学

项目金额： 23万元

中文摘要： 动脉粥样硬化（AS）是糖尿病患者致死致残的主要原因。血管平滑肌细胞的异常增殖是AS发生、发展的重要步骤。内质网应激与AS发生发展过程中血管平滑肌细胞的增殖异常密切相关。糖原合成酶激酶 3β（GSK3β）在细胞增殖调控和内质网介导的细胞凋亡途径中均发挥着重要的作用。本研究以GSK3β基因与内质网应激之间的关系为切入点，研究糖尿病AS发生发展过程中血管平滑肌细胞增殖异常的分子机制。首先建立糖尿病AS小鼠模型，设立GSK3抑制剂组；随后利用基因芯片法分析处理后不同实验组小鼠内质网应激相关基因的表达变化，筛选出候选基因；通过建立血管平滑肌细胞的体外内质网应激细胞模型，结合GSK3β基因过表达和干扰载体的转染，以及免疫共沉淀、启动子荧光素酶检测系统等相关分子生物学检测方法明确GSK3β基因的具体调控机制，以期为研究GSK3β和内质网应激在糖尿病AS形成过程中的作用机制，寻找新的作用靶点提供理论依据。

中文关键词： 糖尿病动脉粥样硬化；GSK3β抑制剂；血管平滑肌细胞；细胞增殖调控；上皮间质转化

英文摘要： Diabetes is an independent risk factor for atherosclerosis-the second cause of death in the world. Understanding the molecular and cellular mechanisms by which diabetes mellitus promotes atherosclerosis(AS) provides a promising hypothesis and multiple new targets for potential therapeutic intervention in the treatment of diabetes mellitus and accelerated AS. The goal of this study is to identify a novel mechanism involving GSK3β and endoplasmic reticulum stress pathways as a potential link between hyperglycaemia and AS. The diabetic AS model group, GSK3β inhibitor treated group and the normal control group were established. Biochemical and slice staining analysis were taken to detect the effect of GSK3β inhibitor in the development of diabetic AS and the proliferation activity of vascular smooth muscle cells(VSMCs). The gene chip is used to screen the target genes of GSK3β. To identify the specific regulation mechanism between GSK3β and ERS pathways, we construct the overexpression and RNAi recombinant expression vector with GSK3β. Then, the transfected cells and normal cells were treated with Tunicamycin to induce ERS cell model, and the sample were detected by immunoprecipitation, luciferase reporter system, Real time-PCR and western blot to determine the target site of GSK3β in endoplasmic reticulum stress p

英文关键词： diabetic atherosclerosis；GSK3β inhibitor；vascular smooth muscle cells；regulation of cell proliferation；epithelial-mesenchymal transition