RhoA对AngⅡ诱导的血管外膜成纤维细胞增殖的影响  

Effect of RhoA on proliferation of adventitial fibroblasts induced by angiotensinⅡ

作　　者：楚玉峰[1] 陈闻东[1] 朱鼎良[1] 高平进[1] 

机构地区：[1]上海交通大学医学院瑞金医院上海市高血压研究所血管生物学重点实验室基因组学国家重点实验室,上海200025

出　　处：《上海交通大学学报（医学版）》2007年第10期1181-1184,共4页Journal of Shanghai Jiao tong University:Medical Science

基　　金：国家重点基础研究发展规划项目("九七三"项目)(2006B503804);上海市科委基金(05JC14038);上海市教委基金(06BZ003)~~

摘　　要：目的构建dominant negative N19RhoA重组腺病毒并感染血管外膜成纤维细胞(AF),观察RhoA对血管紧张素Ⅱ(AngⅡ)诱导的AF增殖的影响。方法运用pAdEasy-1腺病毒载体系统,细菌同源重组法构建含目的基因N19RhoA的腺病毒重组质粒pAd-CMV-N19RhoA-eGFP,并感染体外培养的大鼠AF,然后用1×10-7mol/L的AngⅡ处理24 h。Rho pull-down分析法测定RhoA活性;BrdU ELISA法检测AF的增殖情况。以同时构建的pAd-CMV-eGFP作为对照。结果1×10-7mol/L AngⅡ处理感染pAd-CMV-N19 RhoA-eGFP的AF 30 min后,其GTP-RhoA的表达明显升高,提示AngⅡ可激活RhoA信号分子;感染pAd-CMV-N19 RhoA-eGFP的AF,AngⅡ诱导的BrdU掺入明显受到抑制。结论RhoA信号分子对AngⅡ诱导的AF增殖具有调节作用。Objective To construct dominant-negative N19RhoA recombinant adenovirus with which adventitial fibroblasts （AFs） are infected, and to investigate the effect of RhoA on the proliferation of AFs induced by angiotensin Ⅱ （ Ang Ⅱ ）. Methods Dominantnegative N19RhoA adenovirus recombinant plasmid pAd-CMV-N19RhoA-eGFP was constructed by using pAdEasy-1 adenovirus vector system. The rat AFs cuhured in vitro were infected, and treated with Ang Ⅱ of 1×10^-7 mol/L for 24 h. RhoA activity was determined by Rho pull-down analysis, and AF proliferation by BrdU ELISA. The pAd-CMV-eGFP constructed at the same time was served as control. Results Thirty min after treatment with 1 × 10^-7 mol/L Ang Ⅱ , the expression of RhoA of pAd-CMV-N19RhoA-eGFP-infected AF was significantly increased, indicating that RhoA signaling molecules might be activated by Ang Ⅱ. The BrdU incorporation of AFs infected by pAd-CMV-N19RhoA-eGFP induced by Ang Ⅱ was significantly inhibited. Conclusion RhoA signaling molecules may be involved in AF proliferation induced by Ang Ⅱ.

关 键 词：血管紧张素Ⅱ RHOA 血管外膜成纤维细胞 增殖 腺病毒 

分 类 号：Q78[生物学—分子生物学]