内皮素-1调控SOCC/TGF-β参与房颤大鼠发生心房纤维化  

Endothelin-1 regulates SOCC/TGF-βand involved in atrial fibrosis in rats with atrial fibrillation

作　　者：贾卓然 代曼玉 梁士楚 吴健 薛杨诚 张定欣 沈兵 赵韧[1] Jia Zhuoran;Dai Manyu;Liang Shichu;Wu Jian;Xue Yangcheng;Zhang Dingxin;Shen Bing;Zhao Ren(Dept of Cardiology,The First Affiliated Hospital of Anhui Medical University,Hefei 230022;Dept of Cardiology,West China Hospital,Sichuan University,Chengdu 610041;Dept of Physiology,School of Basic Medical Science,Anhui Medical University,Hefei 230032)

机构地区：[1]安徽医科大学第一附属医院心内科,合肥230032 [2]四川大学华西医院心脏内科,成都610041 [3]安徽医科大学基础医学院生理学教研室,合肥230032

出　　处：《安徽医科大学学报》2024年第3期429-435,共7页Acta Universitatis Medicinalis Anhui

基　　金：国家自然科学基金(编号:81970446)。

摘　　要：目的探讨内皮素-1(ET-1)对心房颤动(AF)大鼠心房纤维化的作用和机制。方法14只成年雄性SD大鼠随机分配为对照(NC)组、房颤(AF)组;采用连续1周每日尾静脉注射1次氯化钙-乙酰胆碱混合液0.1 ml/100 g的方法建立AF大鼠模型,NC组同等方式注射等量生理盐水;各组均在第1天及第8天记录窦性或AF心电图,超声心动图监测心房大小和心功能;采用HE染色及Masson染色观察心房组织纤维化情况;采用Western blot法检测心房组织中内皮素-1(ET-1)、Ⅰ型胶原(COL-Ⅰ)、转化生长因子(TGF)-β、钙库操纵性钙通道(SOCC)蛋白Orai1和基质相互作用分子1(STIM1)的表达;培养小鼠心房肌细胞HL-1细胞,以梯度浓度的ET-1处理24 h后,采用Western blot法观察HL-1细胞ET-1/SOCC/TGF-β信号通路中TGF-β、Orai1及STIM1蛋白表达变化;利用siRNA转染方法敲低HL-1细胞中Orai1表达,以合适浓度的ET-1处理细胞24 h,Western blot检测HL-1细胞中TGF-β蛋白的表达情况。结果与NC组相比,超声心动图显示AF大鼠心脏左房内径(LAD)增加(P<0.05);HE和Masson染色结果显示AF组大鼠心房组织纤维化(P<0.05),Western blot检测结果提示AF组心房组织中ET-1、Orai1、STIM1、TGF-β、COL-Ⅰ蛋白表达较NC组增加(P<0.05)。ET-1处理HL-1细胞后,HL-1细胞Orai1、STIM1、TGF-β蛋白表达增多(P<0.05)。敲低HL-1细胞中Orai1表达后,ET-1的处理不再使TGF-β的表达上调。结论AF大鼠心房组织ET-1表达增多,并通过ET-1/SOCC/TGF-β信号通路促进心房纤维化。Objective To investigate the effect and mechanism of endothelin-1(ET-1)on atrial fibrosis in Atrial fibrillation(AF)rats.Methods Fourteen adult male SD rats were randomly divided into normal control(NC)group and Atrial fibrillation(AF)group.The rat model of Atrial fibrillation was established by injecting 0.1 ml/100g CaCl 2-Ach mixture into the tail vein once a day for one week.The control group was injected with the same dose of normal saline.An electrocardiogram of normal or atrial fibrillation was recorded on the first day and the eighth day in each group,and echocardiography was used to monitor atrial size and cardiac function.The fibrosis of atrial was observed using Masson and HE staining.The expression of endothelin-1(ET-1),collagen-I(Col-I),transforming growth factor-β(TGF-β)and the store operated calcium channel(SOCC)protein Orai1,stromal interaction molecule 1(STIM1)in atrial tissue were detected by Western blot.HL-1 cells were cultured and treated with gradient concentration of ET-1 for 24 hours.Western blot was used to observe changes in the expression of TGF-β,Orai1 and STIM1 proteins in ET-1/SOCC/TGF-β signaling pathway of HL-1 cells.Small interfering RNA(siRNA)transfection method was used to knock down the expression of Orai1 in HL-1 cells,then the cells were treated with appropriate concentrations of ET-1 for 24 hours,and the expression of TGF-β protein in HL-1 cells was detected by Western blot.Results Compared with the control group,echocardiography showed a significant increase in left atrial diameter(LAD)of the heart in atrial fibrillation rats(P<0.05).The HE and Masson staining results showed significant fibrosis in the myocardial tissue of AF group rats(P<0.05),and the Western blot results indicated the expression of ET-1,Orai1,STIM1,TGF-βand COL-Ⅰin the myocardial tissue of AF group significantly increased compared to the NC group(P<0.05).After ET-1 treatment of HL-1 cells,the protein expression of Orai1,STIM1and TGF-β increased(P<0.05),while knocking down Orai1 in HL-1 cells,ET-1

关 键 词：内皮素-1 心房颤动 心房纤维化 Orai1 基质相互作用分子1 TGF-Β HL-1细胞 

分 类 号：R541.7[医药卫生—心血管疾病]