钙蛋白酶介导线粒体ROS生成在血管重构过程中的作用及机理

The involvement of calpain and mitochondrial reactive oxygen species (m-ROS) in the progression of pathophysiologies of vascular remodeling has been recognised, however, the role and mechanisms of calpain mediating m-ROS generation in regulating vascular remodeling has never been reported. Our preliminary data have shown that Ang-Ⅱ induced the activation of calpain which correlated with the increase of m-ROS generation in vascular smooth muscle cell（VSMC）.Furthermore, incubation with calpain repressed mitochondrial ATP synthase and induced m-ROS generation.Since m-ROS has been nicely shown to induce vascular remodeling, we hypothesize that calpain mediates m-ROS generation contributing to the pathophysiological progress of vascular remodeling. Accordingly, the present project is proposed to investigate the role of calpain in vascular remodeling and to explore the underlying mechanisms. Specially, we will induce vascular remodeling by Ang-Ⅱ on Calpastatin transgenic mice and study how calapin mediates m-ROS generation leading to vascular remodeling. In cultured VSMC, we will confirm the in vivo findings. The completion of present study will help understand the molecular mechanisms by which calpain mediates m-ROS generation and provide novel therapeutic targets for new drug design to treat the vascular remodeling.

钙蛋白酶（Calpain）和线粒体活性氧（m-ROS）均参与血管重构的病理生理过程，然而在血管重构过程中关于二者之间调控关系的的机制研究却未见报道。我们的前期研究发现Ang-Ⅱ刺激后血管平滑肌细胞内Calpain发生激活并且与m-ROS生成密切相关，Calpain与分离的线粒体共孵育能够抑制线粒体ATP合成酶的表达、促进m-ROS生成，据此推测调控m-ROS生成是Calpain参与血管重构过程的主要机制之一。本课题拟利用Calpastatin转基因小鼠构建血管重构模型，观察Calpain对m-ROS的调控作用，进一步在细胞水平揭示Calpain通过调节ATP合成酶进而干预m-ROS生成的分子机制。本课题通过阐明Calpain与m-ROS生成的关系，以拓展对血管重构发生发展的分子调控机制的认识，为有效干预和治疗血管重构的药物研发提供新的思路。

钙蛋白酶（Calpain）和线粒体活性氧（m-ROS）均参与血管重构的病理生理过程，然而在血管重构过程中关于二者之间调控关系的的机制研究却未见报道。我们的前期研究发现Ang-Ⅱ刺激后血管平滑肌细胞内Calpain发生激活并且与m-ROS生成密切相关，Calpain与分离的线粒体共孵育能够抑制线粒体ATP合成酶的表达、促进m-ROS生成，据此推测调控m-ROS生成是Calpain参与血管重构过程的主要机制之一。本课题利用Calpastatin转基因小鼠构建血管重构模型，观察Calpain对m-ROS的调控作用，进一步在细胞水平揭示Calpain通过调节ATP合成酶进而干预m-ROS生成的分子机制。本课题通过阐明Calpain与m-ROS生成的关系，以拓展对血管重构发生发展的分子调控机制的认识，为有效干预和治疗血管重构的药物研发提供新的思路。另外，我们探讨了Apelin-13在脂多糖（Lipopolysaccharide, LPS）诱导的血管内皮细胞凋亡中的作用及其可能的机制，以及Apelin对高糖诱导的原代人脐静脉内皮细胞（Human Umbilical Vein Endothelial Cells，HUVECs）中ROS生成和糖尿病小鼠内皮依赖性血管功能障碍的影响。研究结果发现Apelin能够抑制高糖诱导的血管内皮细胞活性氧生成和细胞凋亡、上调NO的含量、减轻糖尿病小鼠血管内皮功能障碍，证实Apelin/APJ系统通过拮抗LPS诱导的细胞内ROS水平升高抑制HUVECs凋亡，发挥保护作用。