TRPCs介导PMVECs表型转变在肺纤维化中的作用及机制研究

项目名称： TRPCs介导PMVECs表型转变在肺纤维化中的作用及机制研究

项目编号： No.81500050

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： 颜林枫

作者单位： 中国人民解放军第四军医大学

项目金额： 17万元

中文摘要： 肌成纤维细胞（MFb）过度分泌细胞外基质是特发性肺纤维化的病理基础，而转化生长因子β1（TGF-β1）和结缔组织生长因子（CTGF）是促进MFb形成的关键细胞因子。我们研究发现，博莱霉素（BLM）诱导的肺纤维化大鼠肺微血管内皮细胞（PMVECs）持续高表达TGF-β1和CTGF，细胞呈增殖表型且[Ca2+]i过载，但原因不明。瞬时受体电位通道C型（TRPCs）是内皮细胞表面重要的钙离子通道，其介导的 [Ca2+]i增加参与调节内皮细胞增殖和分化，但肺纤维化时PMVECs表型改变是否与TRPCs活性调节有关及其作用机制尚不清楚。本课题拟以TRPCs介导的钙代谢异常为切入点，采用免疫组化、蛋白印迹、激光共聚焦及药物干预等方法，从细胞及活体研究BLM作用后PMVECs表型转变与TRPCs的相关性，探讨PMVECs表型转变在肺纤维化中的作用及其机制，为确定肺纤维化早期干预的靶点提供实验和理论基础。

中文关键词： 肺纤维化；肺微血管内皮细胞；钙离子；瞬时受体电位通道C型；表型

英文摘要： The pathological basis of idiopathic pulmonary fibrosis is the accumulation of large amounts of extracellular matrix produced by Myofibroblasts(MFbs) in interstitial lung. The appearance of MFbs is mainly induced by TGF-β1 and CTGF. Our previous research demonstrated that TGF-β1 and CTGF were persistent highly expressed in pulmonary microvascular endothelial cells (PMVECs) of lung fibrosis induced by BLM, which were proliferative phenotype and overloaded with [Ca2+]i, but for unknown reasons. Transient receptor potential channels canonical(TRPCs) are important Ca2+ channels, which are involved in the regulation of endothelial cells’ proliferation and differentiation by mediating increased [Ca2+]i. However, it is unknown whether PMVECs’ phenotypic change in lung fibrosis is related to the activity of TRPCs. In present study, regarding abnormal calcium metabolism mediated by TRPCs as breakthrough point, we aimed to study the relationship between TRPCs and PMVECs' phenotypic change using the methods of immunohistochemistry, western blotting, confocal laser scanning and drug intervention, and to explore the role of PMVECs' phenotypic change and its mechanism in lung fibrosis, in order to provide experimental and theoretical basis in determining targets for the early intervention of lung fibrosis.

英文关键词： pulmonary fibrosis;pulmonary microvascular endothelial cells;calcium;Transient receptor potential channels canonical;phenotype