RhoA/Rho激酶通路在血管紧张素Ⅱ诱导大鼠血管外膜成纤维细胞表达纤维连接蛋白中的作用  

The role of RhoA/Rho kinase pathway on fibronectin expression in rat vascular adventitial fibroblasts induced by angiotensin Ⅱ

作　　者：楚玉峰[1] 陈闻东[2] 周海燕[3] 蒋进皎[1] 何晓鹏[1] 赵鹏[1] 王春亭[1] 任宏生[1] 

机构地区：[1]山东大学附属省立医院MICU,济南山东250021 [2]上海交通大学医学院附属瑞金医院,上海市血管生物学重点实验室 [3]山东省肿瘤医院肿瘤内科

出　　处：《中华高血压杂志》2015年第8期743-748,共6页Chinese Journal of Hypertension

基　　金：国家自然科学基金(81200238;81200176);山东省优秀中青年科学家科研奖励基金项目(BS2011YY043;BS2011YY052);山东省自然科学基金(ZR2013HM062);山东省科技发展计划(2014GSF118018)

摘　　要：目的观察血管紧张素Ⅱ(AngⅡ)对大鼠血管外膜成纤维细胞(VAF)纤维连接蛋白(FN)表达的影响,并探讨RhoA/Rho激酶通路在AngⅡ诱导FN表达中的相关机制。方法分别以不同时间(0、6、12、24、48h)和不同剂量(0、10-8、10-7、10-6、10-5 mol/L)AngⅡ刺激体外培养的SD大鼠VAF,应用实时荧光定量逆转录聚合酶链反应及Western blot检测FN的表达变化,并分别应用表达目的基因N19RhoA的重组腺病毒和Rho激酶抑制剂Y27632进行干预,观察其对AngⅡ诱导FN表达的影响。结果 AngⅡ促进大鼠VAF表达FN。应用10-7 mol/L AngⅡ刺激VAF 24h,FN mRNA表达和蛋白表达增高最为明显(均P<0.05)。与AngⅡ刺激组比较,表达目的基因N19RhoA的重组腺病毒和Y27632可明显抑制AngⅡ诱导的FN mRNA(N19RhoA组0.78±0.11,Y27632组0.72±0.08比AngⅡ刺激组1.82±0.12,均P<0.05)和蛋白表达(N19RhoA组1.56±0.12,Y27632组1.43±0.12比AngⅡ刺激组3.89±0.32,均P<0.05)。结论 AngⅡ促进大鼠VAF表达FN,RhoA/Rho激酶通路参与了该过程的调控。Objective To investigate the effects of angiotensin Ⅱ （Ang Ⅱ ） on fibronectin （FN） expression in rat vascular adventitial fibroblasts （VAF） and the role of RhoA/Rho kinase pathway in mediating FN expression. Methods The VAF of SD rats were exposed to different concentrations （0, 10^-8, 10^-7, 10^-6, 10^-5 tool/L） of Ang Ⅱ for various time （0, 6, 12, 24, 48 h） in vitro. Inhibition of RhoA and Rho kinase was achieved by infec- tion with N19RhoA recombinant adenovirus or treated with Y27632. The effects of N19RhoA recombinant adeno- virus and Y27632 on FN expression were determined by real-time quantitative reverse transcription polymerase chain reaction and Western blot. Results Ang Ⅱ induced FN expression in VAF. The mRNA and protein levels of FN in VAF rose significantly after exposure to Ang Ⅱ at 10^-7 mol/L for 24 h （all P〈0.05）. The N19RhoA recombi- nant adenovirus and Y27632 markedly attenuated the upregulation of FN induced by Ang Ⅱ （mRNA： 0.78±0.11 in N19RhoA cells, 0.72±0.08 in Y27632 cells vs 1.82±0.12 in Ang Ⅱ cells; protein： 1.56±0.12 in N19RhoA cells, 1.43±0.12 in Y27632 cells vs 3.89±0.32 in Ang Ⅱ cells, all P〈0.05）. Conclusion FN was upregulated in VAF induced by Ang Ⅱ , and the RhoA/Rho kinase pathway was involved in FN expression mediated by Ang Ⅱ.

关 键 词：血管紧张素Ⅱ 血管外膜成纤维细胞 RhoA/Rho激酶通路 纤维连接蛋白 

分 类 号：R54[医药卫生—心血管疾病]