姜黄素和氯沙坦对血管紧张素Ⅱ诱导的内皮间质细胞转化的作用及机制研究  

The effects and mechanisms of curcumin and losartan on angiotensin Ⅱ-induced endothelial-mesenchymal transition

作　　者：胡丐锋 黄裕立[1] 黄伟俊[1] 黎文生[1] 麦林琳[1] 童辉煜 李美君[1] 程宏基 申常造 胡允兆[1] 

机构地区：[1]南方医科大学附属顺德第一人民医院心内科,广东佛山528300

出　　处：《广东医学》2017年第6期821-824,共4页Guangdong Medical Journal

基　　金：国家自然科学基金青年科学基金资助项目(编号:81600239)

摘　　要：目的研究姜黄素和氯沙坦对血管紧张素(Ang)Ⅱ诱导的内皮间质细胞转化(End MT)的作用及机制。方法培养人脐静脉内皮细胞(HUVECs)然后分为4组:对照组(不加刺激药物的正常培养组);AngⅡ组(加入刺激浓度为0.1μmol/L或1μmol/L的AngⅡ);姜黄素+AngⅡ组(先加入12.5μmol/L的姜黄素预孵1 h,再加入1μmol/L的AngⅡ);氯沙坦+AngⅡ组(先加入10μmol/L的氯沙坦孵育2 h,再加入1μmol/L的AngⅡ);刺激24 h后行划痕试验检测各组细胞迁移能力改变和CCK-8检测细胞活力;刺激5 d后,显微镜观察各组细胞形态变化和Western blot检测各组血管内皮钙黏蛋白(VE-cadherin)、α-平滑肌肌动蛋白(α-SMA)、转化生长因子β_1(TGF-β_1)蛋白表达量。结果 HUVECs经AngⅡ刺激后形态改变,长梭形样细胞明显增多;姜黄素和氯沙坦可维持内皮细胞铺路石样形态。AngⅡ呈浓度依赖性地降低HUVECs存活率(P<0.01);各组存活率均>50%(均P<0.05)。相比对照组,AngⅡ促进HUVECs的迁移(P<0.05);相比AngⅡ组,加入姜黄素和氯沙坦可抑制HUVECs的迁移(均P<0.05)。AngⅡ可诱导EndMT,AngⅡ刺激5 d后VE-cadherin表达减弱(P=0.003),而α-SMA和TGF-β_1的表达都明显增强(均P<0.01),姜黄素和氯沙坦使VE-cadherin表达明显增加,而α-SMA和TGF-β_1的表达均明显下调。结论姜黄素和氯沙坦通过下调TGF-β_1表达抑制AngⅡ诱导的EndMT。Objective To investigate the effects of curcumin （Cur） on angiotensin - Ⅱ （Ang Ⅱ ） - induced en- dothelial - mesenchymal transition （EndMT） and the mechanisms underlying the anti - fibrotic effect of Cur and losartan. Methods Primary human umbilical vein endothelial cells （HUVECs） were cultured and divided into four groups, normal control group, Ang Ⅱ group, Cur plus Ang Ⅱ group, and losartan plus Ang Ⅱ group. The morphological changes were recorded by microscope. Cell viability was measured by CCK8 assay; the migration ability was tested by Scratch -Wound assay; and Western blot method was used to test the expression of VE - cadherin, ~ - SMA, and TGF - 61. Results Cellular morphology was significantly different after Ang I1 treatment, showing more spindle - shaped fibroblast pheno- types. However, Cur and losartan blocked the effect of Ang Ⅲ treatment, and the cell morphology was similar to control cells. CCK8 showed that Ang Ⅱ inhibited HUVECs viability in a dose - dependent manner. The viabilities of each group were greater than 50% （ P 〈 0.05 ）. Scratch - wound assay suggested that Ang Ⅱ significantly promoted the migration ability （P =0. 044 for0. 1 μmol/L Ang Ⅲ group, P 〈0. 01 for μmol/L Ang]] group）, which were reversed by Cur and losartan （ both P 〈 0. 05 ）. Pretreatment of HUVECs with Cur and losartan inhibited Ang Ⅱ - induced expression of α - SMA and TGF - β1, and increased the expression of VE - cadherin. And there was no significant difference in the effects of Cur and losartan on α - SMA, TGF - β1, or VE - cadherin （P 〉 0. 05 ）. Conclusion Cur and losartan inhibit Ang Ⅱ - induced aberrant EndMT by down - regulating TGF - β1 expression.

关 键 词：姜黄素 氯沙坦 内皮间质细胞转化 血管紧张素Ⅱ 

分 类 号：R285.5[医药卫生—中药学]