同型半胱氨酸经ERK通路上调ETB受体表达促血管平滑肌细胞增殖机制

项目名称： 同型半胱氨酸经ERK通路上调ETB受体表达促血管平滑肌细胞增殖机制

项目编号： No.81500350

项目类型： 青年科学基金项目

立项/批准年度： 2016

项目学科： 医药、卫生

项目作者： 陈玉龙

作者单位： 西安医学院

项目金额： 18万元

中文摘要： 内皮素B（ETB）受体促血管平滑肌细胞（VSMC）增殖作用是促进动脉粥样硬化（As）发生发展的重要机制之一。预实验结果显示，同型半胱氨酸（Hcy）上调VSMC中ETB受体表达与亚硝基化ERK水平减少有关，但机制不明。本研究拟利用VSMC和离体血管模型，明确Hcy对VSMC中亚硝基化ERK、磷酸化ERK、ETB受体表达水平及细胞增殖的影响；利用磷酸化ERK、ETB受体抑制剂或过表达亚硝基化ERK模拟物的腺病毒处理模型，分别阻断ERK磷酸化和ETB受体表达，或恢复亚硝基化ERK水平，探讨Hcy通过亚硝基化ERK/磷酸化ERK/ETB受体途径促VSMC增殖的机制；检测ERK下游NF-κB通路关键蛋白磷酸化水平以及NF-κB的活性，进一步探讨ERK调控ETB受体的下游作用机制。利用动物模型，对体外实验结果进行在体验证。这将为阐明Hcy致As的发病机制和探讨As防治提供新的实验和理论依据。

中文关键词： 动脉粥样硬化；同型半胱氨酸；内皮素B型受体；细胞外信号调节激酶；亚硝基化

英文摘要： Vascular smooth muscle cell (VSMC) proliferation induced by endothelin B (ETB) receptor could promote development of atherosclerosis. Our data showed that homocysteine(Hcy) up-regulated expression of ETB receptor, accompany with reduction of S-nitrosylated ERK in VSMC. Relationship between them is unclear. We will determine effect of Hcy on levels of S-nitrosylated ERK, phosphorylated ERK, ETB receptor, and cell proliferation in VSMC and vessel culture model, respectively. In order to understand the role of S-nitrosylated ERK/ phosphorylated ERK/ETB receptor pathway, we will examine the effects of phosphorylated ERK inhibitor, ETB receptor or adenovirus overexpressing S-nitrosylation-mimetic ERK protein on VSMC proliferation induced by Hcy. We will determine levels of phosphorylation and activity of NF-κB to investigate mechanism of ERK regulating ETB receptor in VSMC and vessel culture model treated by Hcy. Finally, we will further confirm the results of in vitro experiments using animal models. This study will provide new experimental and theoretical basis for clarifying mechanism of atherosclerosis induced by Hcy, and investigating prevention of atherosclerosis.

英文关键词： Atherosclerosis;Homocysteine;Endohelin B type receptor;Extracellular signal-regulated kinase;S-nitrosylation