甲羟戊酸途径的干预对自发性高血压大鼠内皮功能的影响

项目名称： 甲羟戊酸途径的干预对自发性高血压大鼠内皮功能的影响

项目编号： No.81200191

项目类型： 青年科学基金项目

立项/批准年度： 2013

项目学科： 医学一处

项目作者： 韩杰

作者单位： 浙江大学

项目金额： 23万元

中文摘要： 本项目拟在前期发现自发性高血压大鼠（SHR）心血管组织中甲羟戊酸途径的非固醇类分子途径关键酶的表达上调的基础上，进一步观察甲羟戊酸途径的干扰是否可以逆转失衡的NO-ROS途径从而改善SHR受损的内皮功能。为明确其机制，拟开展下述研究：（1）在体SHR动物实验和体外内皮细胞、平滑肌细胞实验中采用化学抑制剂或shRNA抑制上调的FDPS，FNTA, FNTB，GGTase-I等酶, 进一步观察血清/细胞NO、ROS水平；血管/细胞NOS、NADPH氧化酶、抗氧化酶的活性及各亚基mRNA与蛋白表达、磷酸化水平；（2）观察血管/细胞甲羟戊酸途径下游活性分子小G蛋白RhoA、Rac1等相关蛋白表达和活性；（3）离体灌流实验检测血管功能。以进一步明确小G蛋白在高血压内皮功能损伤过程中的作用及甲羟戊酸途径影响内皮功能的上、下游分子信号转导机制。

中文关键词： 甲羟戊酸途径；活性氧；烟酰胺腺嘌呤二核苷酸磷酸氧化酶；小G蛋白Rac1；内皮功能失调

英文摘要： In previous studies, we found that the expression of key enzymes in the mevalonate pathway was significantly unregulated in spontaneously hypertensive rat (SHR). The purpose of this study is to determine the role of small G protein, such as RhoA and Rac1, in endothelial dysfunction and whether mevalonate pathway inhibition improves the endothelial function in SHR, through reversing the imbalance of NO-ROS pathway. To investigate the underlying mechanisms, we carry out the following studies. SHR rats or endothelial cells, smooth muscle cells in vitro were treated with chemical inhibitors or shRNA of over-expressioned key enzymes including FDPS，FNTA, FNTB，GGTase-I. Then the production of NO and ROS, activity of NOS, NADPH oxidase and antioxidant enzyme were determined by fluorescence. The mRNA level, protein expression and phosphorylation level of NOS, NADPH oxidase and antioxidant enzyme were determined by Real-time PCR and western blot, respectively. The protein expression and activity of RhoA / Rac1 in downstream of mevalonate pathway were determined by western blot and fluorescence. The vascular function was detected by isolated perfused assay. Our study may open the field for novel therapeutic indications and give many possible therapeutic targets beyond arterial hypertension in the future.

英文关键词： mevalonate pathway；reactive oxygen species；NADPH oxidase；Rac1；endothelial dysfunction