血管紧张素Ⅱ通过ERK1/2通路促进血管内皮细胞钙化  

The mechanism of angiotensin Ⅱ regulates the calcification in HUVECs through ERK1/2 pathway

作　　者：程治源 叶挺 凌秋洋 吴婷[2] 马涛 王倩[1] 宗刚军[1,2] Cheng Zhiyuan;Ye Ting;Ling Qiuyang(Dept of Cardiology, Wuxi Clinical Hospital, Anhui Medical University, Wuxi 214044;Dept of Cardiology ,101 Hospital of PLA, Wuxi 214044)

机构地区：[1]安徽医科大学无锡临床学院心血管内科,无锡214044 [2]解放军第101医院心血管内科,无锡214044

出　　处：《安徽医科大学学报》2018年第4期542-546,共5页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81371657);无锡市卫生计生委重大科研项目(编号:z201608)

摘　　要：目的探讨血管紧张素II通过ERK1/2通路促进血管钙化的病理生理学机制。方法体外培养HUVECs 5 d后,用不同浓度梯度血管紧张素Ⅱ刺激内皮细胞,筛选出诱导血管内皮细胞钙化的最适浓度。用血管紧张素Ⅱ刺激内皮细胞不同时间点(0、15、30、45、60 min),检测细胞外信号调节激酶1/2(ERK1/2)通路磷酸化水平。用血管紧张素Ⅱ诱导5 d的钙化内皮细胞分为4组:对照组(用5%胎牛血清的ECM培养液)、血管紧张素Ⅱ组(100 nmol/L AngⅡ)、氯沙坦组(100 nmol/L AngⅡ+1μmol/L Losartan)、U0126组(100 nmol/L AngⅡ+U01261μmol/L)。通过Western blot、ELISA检测骨形态发生蛋白-2、4(BMP2、BMP4)钙化因子表达来探讨血管紧张素Ⅱ对血管内皮钙化的影响及其信号通路。结果血管紧张素Ⅱ增加人脐静脉内皮细胞(HUVECs)的BMP2、BMP4钙化因子表达(P<0.05);而血管紧张素Ⅱ1型受体阻断剂和ERK1/2通路抑制剂U0126可下调血管紧张素Ⅱ对血管内皮BMP2、BMP4的表达(P<0.05)。结论血管紧张素Ⅱ可诱导HUVECs钙化,其途径是通过ERK1/2通路促进血管内皮细胞钙化。Objective To investigate the pathophysiological mechanism of angiotensin II promoting vascular calcification through ERK1/2 pathway. Methods After cultured for 5 days in vitro,endothelial cells were stimulated with different concentrations of angiotensin Ⅱ,and the optimal concentration of angiotensin Ⅱ for inducing calcification of vascular endothelial cells was screened out. The cells were randomly divided into four groups and treated with angiotensin Ⅱ（ Ang Ⅱ） according to the following： Control,Ang Ⅱ（ 100 nmol/L）,Ang Ⅱ ＋ U0126（ AngⅡ 100 nmol/L ＋ U0126 1 μmol/L）,Ang Ⅱ ＋ Losartan（ Ang Ⅱ 100 nmol/L ＋ Losartan 1μmol/L）. The expression of BMP2 and BMP4 was detected by Western blot and ELISA to explore the effect of angiotensin Ⅱ on vascular endothelial calcification and its signal pathway. Results Ang Ⅱ treatment of HUVECs led to significant up– regulation of BMP2 and BMP4 expression and activation. Neither the BMP2 or BMP4 proteins,nor the secreted proteins were detectabled in the control group or Losartan treated group or U0126 treated group; however,there was a significant increase in Ang Ⅱ-treated group（ P〈 0. 05）. The BMP2 and BMP4 proteins were not expressed in the control or Losartan treated groups or others,whereas in Ang Ⅱ – treated cells both BMP2 and BMP4 proteins expression were enhanced（ P 〈0. 05）. The effect of Ang Ⅱ could be inhibited by the addition of Losartan（ P〈 0. 05）. Conclusion Ang Ⅱ promotes calcification of HUVECs through ERK1/2 pthway,which lead to pathological vascular calcification.

关 键 词：血管紧张素Ⅱ 人脐静脉内皮细胞 钙化 ERK1/2通路 

分 类 号：R561.2[医药卫生—呼吸系统]