miR-124调节TNFα介导平滑肌细胞胶原合成酶P4Hα1表达机制及对斑块易损性的作用

项目名称： miR-124调节TNFα介导平滑肌细胞胶原合成酶P4Hα1表达机制及对斑块易损性的作用

项目编号： No.81470559

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： 张梅

作者单位： 山东大学

项目金额： 68万元

中文摘要： 斑块炎症、胶原纤维含量与斑块易损性有关，P4Hα1是胶原合成过程的关键酶。炎症因子对胶原合成和斑块稳定性研究甚少，且机制不明确。我们研究发现在易损斑块中miR-124表达显著升高，P4Hα1表达降低；TNFα抑制P4Hα1的表达，P4Hα1过表达可增加胶原含量和斑块稳定性，但其作用机制尚需研究。为此我们提出假说：TNFα经NF-Y/FOXP1增加miR-124表达，miR-124可通过降低P4Hα1蛋白表达，降低斑块胶原蛋白含量而增加斑块易损性。本课题将从体内外研究验证这一假说，利用RT-PCR、Western blot、慢病毒载体转染、荧光素酶报告基因、融合蛋白、EMSA、CHIP及miRNA等技术，从分子、细胞、组织以及动物整体水平等探讨miR-124在炎症影响胶原合成和斑块稳定性过程中的作用和分子机制。本研究将从胶原合成角度揭示易损斑块发生机制，为稳定斑块的干预策略提供新的思路。

中文关键词： 易损斑块；miR-124；胶原合成

英文摘要： Collagen plays an important role in plaque stabilization. Inflammation is one of the main modulators of collagen metabolism. However, studies of inflammation-induced plaque destabilization were mainly focused on collagen degradation. Little is known about the role the other part of collagen metabolism, collagen synthesis, during the process. P4Hα1 is a key enzyme during synthesis of collagens. Our preliminary experiments showed that in destabilized plaques, P4Hα1, together with Collagen I and III, was significantly suppressed, whereas miR-124 was obviously upregulated. We preconceived that miR-124 suppress P4Hα1 and subsequently Collagen I and III to destabilize plaques with the presence of inflammation factors, such as TNFα. We will investigate the mechanisms involved in miR-124-mediated modulation of collagen synthesis and plaque stability by TNFα both in vitro and in vivo.RT-RCR,Western Blot，transfection of lenti-virus, luciferase reporter gene assay, EMSA, CHIP, miRNA detection and analysis techniques will be applied. The study will include molecular and cellular experiments, organic experiments and animal experiments. We hope that our research will contribute to the understanding of the role of collagen synthesis in plaque destabilization and provide new insight into prevention and treatment of vulnerable plaques and acute cardiovascular events.

英文关键词： vulnerable plaque;miR-124;collagen synthesis