ACE2小干扰RNA对平滑肌细胞AT1受体蛋白表达及其下游信号通路的影响  

Expression of AT1 receptor protein and its downstream signaling pathways in vascular smooth muscle cells after ACE2 siRNA interference

作　　者：龚晶婧[1] 李荣通 卢卓强[1] 许昌声 王华军 晋学庆[3] GONG Jing-jing;LI Rong-tong;LU Zhuo-qiang;XU Chang-sheng;WANG Hua-jun;JIN Xue-qing(Dept of Cardiology,the First Affiliated Hospital,Fujian Medical University,Fuzhou 350004,China;The Hypertension Research Division,Fuzhou 350004,China;Cadre Ward,the First Affiliated Hospital of Fujian Medical University,Fuzhou 350004,China)

机构地区：[1]福建医科大学附属第一医院心内科,福州350004 [2]福建省高血压研究所,福州350004 [3]福建医科大学附属第一医院干部病房,福州350004

出　　处：《中国药理学通报》2020年第3期354-359,共6页Chinese Pharmacological Bulletin

基　　金：福建医科大学教授学术发展基金(No JS14023);福建医科大学科研发展基金项目苗圃基金(No 2014MP026)。

摘　　要：目的研究ACE2 siRNA干预SD大鼠平滑肌细胞(vascular smooth muscle cells,VSMCs)后,探讨ACE2对血管紧张素Ⅱ1型受体(AngⅡtype 1 receptor,ATlR)蛋白表达、ERK1/2、STAT3蛋白磷酸化水平的影响。方法(1)用脂质体法将含有ACE2基因的质粒DNA及si-ACE2转染到各对应干预组VSMCs;(2)Western blot检测各组细胞ACE2、AT1R蛋白表达水平以及p-ERK1/2、p-STAT3蛋白的磷酸化水平。结果(1)对比空白对照组、GFP组、脂质体组,si-ACE2组、AngⅡ组ACE2蛋白表达明显降低,而AT1R蛋白表达和p-ERK1/2、p-STAT3蛋白磷酸化水平明显增加(P<0.05);(2)分别对比AngⅡ组和AngⅡ+ACE2组,AngⅡ+si-ACE2组和AngⅡ+ACE2+si-ACE2相应的ACE2蛋白表达均降低,而AT1R蛋白表达、p-ERK1/2、p-STAT3蛋白磷酸化水平均增加(P<0.05)。结论ACE2 siRNA和AngⅡ均能抑制ACE2蛋白的表达,且两者对ACE2起协同抑制作用;ACE2蛋白表达的减少能上调AT1受体蛋白表达,同时提高其下游信号通路ERK1/2、STAT3蛋白磷酸化水平。Aim To investigate the effect of angiotensinⅡtype 1 receptor(AT1R)expression and the level of extracelluar signal-regulated kinase(ERK1/2)and signal transducer and activator of transcription 3(STAT3)phosphorylation with the less expression of ACE2 protein after ACE2 siRNA interference in vascu-lar smooth muscle cells(VSMCs).Methods In cor-responding intervention groups the SD rat vascular smooth muscle cells(VSMCs)were transfected by pm-ACE2 and si-ACE2 through lipid method.ACE2,AT1R protein expression,p-ERK1/2 and p-STAT3 protein phosphorylation level were detected by Western blot.Results Western blot showed that the amount of ACE2 protein in si-ACE2 group and AngⅡgroup was less than that in control group,lipofectamine 2000 group and GFP group(P<0.05).In contrast,the AT1R protein expression and the level of p-ERK1/2,p-STAT3 phosphorylation in si-ACE2 group and AngⅡgroup were much higher than those in control group,li-pofectamine 2000 group and GFP group(P<0.05).Compared to AngⅡgroup,the AngⅡ+si-ACE2 group ACE2 protein expression was also significantly lower(P<0.05),and the AT1R protein expression and the level of p-ERK1/2,p-STAT3 phosphorylation were much higher(P<0.05).Conclusions The expres-sion of ACE2 gene in VSMCs could be significantly in-hibited by siRNA and AngⅡ.Besides,si-ACE2 and AngⅡcan synergistically inhibit the expression of ACE2 protein.The decrease of ACE2 protein expres-sion can uP-regulate AT1 receptor expression and in-crease the phosphorylation level of p-ERK1/2 and p-STAT3 proteins in its downstream signaling pathway.

关 键 词：血管紧张素转换酶2 血管紧张素Ⅱ 血管紧张素Ⅱ1型受体蛋白表达 细胞外信号调节激酶1/2蛋白 信号转导和转录激活因子3 小干扰RNA 

分 类 号：R-332[医药卫生] R322.74