基于ERK1/2通路MicroRNAs调控探讨泽泻汤对AS VSMC迁移增殖的机制研究

项目名称： 基于ERK1/2通路MicroRNAs调控探讨泽泻汤对AS VSMC迁移增殖的机制研究

项目编号： No.81473744

项目类型： 面上项目

立项/批准年度： 2015

项目学科： 医药、卫生

项目作者： 薛偕华

作者单位： 福建中医药大学

项目金额： 72万元

中文摘要： 动脉粥样硬化（AS）严重危害人类健康，与脂质沉积和炎性反应相关，巨噬细胞和血管平滑肌细胞（VSMC）在其中起着重要作用。泽泻汤出自《金匮要略》，在临床上广泛用于治疗高脂血症、AS等，本项目前期研究提示泽泻汤能改善巨噬细胞源性泡沫细胞的脂质沉积，抑制炎性反应，但是泽泻汤对AS VSMC的作用及其机制尚未见报道。近来研究表明ERK1/2通路的MicroRNAs（MiRNA-143、145、 29b、 31）与VSMC迁移增殖密切相关。本项目通过泽泻汤含药血清干预Ox-LDL负荷VSMC的AS细胞模型，检测VSMC表型标志蛋白、迁移增殖、MMPs/TIMPs、EMMPRIN、细胞周期等指标，探讨泽泻汤对VSMC迁移增殖的作用，并基于ERK1/2通路MicroRNAs调控深入探索泽泻汤抗AS的作用机制；同时通过扩增泽泻汤靶基因启动子序列，构建启动子-荧光素酶报告基因，验证泽泻汤抗AS的作用靶点。

中文关键词： 动脉粥样硬化；血管平滑肌细胞；泽泻汤；MicroRNAs；迁移增殖

英文摘要： Atherosclerosis(AS) has the great dangerous effects on human health. Lipid deposition, inflammation, and the abnormal secretion of matrix metalloproteinases (MMPs) are closely related with AS. The macrophages and vascular smooth muscle cells (VSMC) play an indispensable role in the process. Our previous studies showed that Alisma Decoction could attenuate lipid deposition and inhibit inflamation in macrophages-derived foam cells. However, the impact of Alisma Decoction on VSMC has not been reported until now.Several studies show that MicroRNAs(MiRNA-143 145 29b 31)related with ERK1/2 singal are associated with VSMC migrant and proliferation. This project utilizes Alisma Decoction to treat Ox-LDL loaded VSMC model. Then we detect the phenotypic transformation marker proteins SM22α and Osteopontin expression levels, cell migrant and proliferation, MMPs/TIMPs, cell cycles and other indicators, aiming to clarify the effect of Alisma Decoction on the changes of VSMC biological behavior, such as phenotype transformation, migration and proliferation . At the same time we explore its effects on anti-AS on MiRNAs levels and its correlation with ERK1/2 pathway, in order to elucidate the mechanisms of Alisma Decoction blocking atherosclerosis.Furthermore we construct the promoter-luciferase by amplification of Alisa Decoction target gene promoter sequences to clarify the Alisa Decoction anti-AS function.

英文关键词： atherosclerosis;vascular smooth muscle cells;Alisma Decoction;MicroRNAs;migration and proliferration