Tbx18参与血管钙化的作用及机制研究

项目名称： Tbx18参与血管钙化的作用及机制研究

项目编号： No.81470488

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： 颜建云

作者单位： 南方医科大学

项目金额： 73万元

中文摘要： 血管钙化常见于心血管疾病，糖尿病和慢性肾病患者，是心血管疾病发生率和死亡率增加的独立危险因素，严重威胁人类健康。Tbx18是调节心血管系统发育的重要转录因子。既往研究发现Tbx18阳性细胞能分化成血管平滑肌细胞。我们的前期实验也表明Tbx18在钙化的血管平滑肌细胞中表达明显增高，且抑制Tbx18能下调血管钙化相关的重要转录因子Runx2和Sox9的表达。由此我们假设：Tbx18可能调节Runx2和Sox9参与血管钙化。本项目拟采用体内外相结合的血管钙化模型，运用腺病毒过表达，RNA干扰及基因敲除的方法，对Tbx18参与血管钙化的作用进行研究；应用RNA-Seq的方法，筛选Tbx18调控的下游基因，然后用染色质免疫共沉淀结合PCR，凝胶迁移，荧光素酶报告基因分析的方法研究转录因子Tbx18与下游基因的相互作用关系，阐明Tbx18调节血管钙化的分子机制，为血管钙化的防治提供新的理论依据和思路。

中文关键词： 动脉粥样硬化；血管平滑肌细胞；血管钙化；骨相关蛋白；Tbx18转录因子

英文摘要： Vascular calcification is very common in a number of diseases including cardiovascular diseases, diabetes, and chronic kidney diseases. It is an independent risk factor for cardiovascular mortality and morbidity and heavily threatens human health. Tbx18 is an important transcription factor regulating cardiovascular development. Previous studies have shown that Tbx18 positive cells give rise to vascular smooth muscle cells during heart development. Our preliminary data also show that Tbx18 is highly expressed in calcified vascular smooth muscle cells, and knock-down of Tbx18 results in down-regulation of vascular calcification-related transcription factors including Runx2 and Sox9. Based on these findings, we hypothesize that Tbx18 may regulate vascular calcification through activation of Runx2 and Sox9. In vitro and in vivo model of vascular calcification will be used in this study. To investigate the role of Tbx18 in vascular calcification, we will use adenovirus encoding Tbx18 and short interfering RNA against Tbx18 to transfect vascular smooth muscle cells. We will also utilize gene knock-out technology in mice to study the function of Tbx18 in vascular calcification. Additionally, we plan to use RNA-Seq to identify potential downstream target genes of Tbx18. Interaction between Tbx18 transcription factor and downstream target genes will be studied by Chip-PCR, electrophoretic mobility shift assay and luciferase reporter analysis. Elucidation of mechanism underlying vascular calcification will provide a novel theoretical basis and treatment for vascular calcification.

英文关键词： atherosclerosis;vascular smooth muscle cells;vascular calcification;bone-related proteins;Tbx18 transcription factor