硫化氢下调Yes相关蛋白1和含PDZ结合基序的转录共激活因子抑制大鼠心肌梗死心肌细胞凋亡及改善心肌纤维化的机制研究  

Hydrogen sulfide inhibits apoptosis of cardiomyocytes and ameliorates myocardial fibrosis in rats after myocardial infarction through downregulating Yes-related protein 1 and a transcriptional co-activator with a PDZ-binding motif

作　　者：汪刘洋 张爱民 赵俊雄 聂连桂 刘盛权 肖婷 杨军[1] Wang Liuyang;Zhang Aimin;Zhao Junxiong;Nie Liangui;Liu Shengquan;Xiao Ting;Yang Jun(Department of Cardiology,The First Affiliated Hospital,University of South China,Hengyang 42100l,China;Department of Cardiology,Longhua District Central Hospital,Shenzhen 518000,China)

机构地区：[1]南华大学第一附属医院心内科,衡阳421001 [2]深圳市龙华区中心医院心内科,深圳518000

出　　处：《中华老年医学杂志》2023年第8期964-969,共6页Chinese Journal of Geriatrics

基　　金：国家自然科学基金(81870230);湖南省自然科学基金优秀青年科学基金项目(2021JJ40499);深圳市龙华区医疗卫生机构区级科研项目(2020035)。

摘　　要：目的探讨外源性硫化氢(H,S)对心肌梗死(心梗)后大鼠心肌纤维化和心肌细胞凋亡的影响及其机制。方法43只SD大鼠按随机数字表法分为4组:对照组12只、心梗组13只、硫化氢组6只和心梗十硫化氢组12只。采用异丙肾上腺素腹腔注射法(50mg/kg,1次/d,共2d)建立急性心梗大鼠模型,末次给药后48h记录心电图及肌钙蛋白变化确定造模情况。造模成功后,心梗组和心梗十硫化氢组大鼠每日腹腔注射硫氢化钠(56μmol/kg,1次/d,持续6周)。6周后比较各组大鼠心脏彩色超声观察心功能变化情况,Masson染色观察各组大鼠心肌胶原容积分数,TUNEL染色检测各组大鼠心肌细胞凋亡率,酶联免疫吸附(Westernblot)法检测Yes相关蛋白1(YAP1)、含有PDZ结合基序的转录共激活因子(TAZ)、哺乳动物STE20样激酶2(MST2)、Bcl-2相关X蛋白(Bax)、胱天蛋白酶3(Caspase-3)、基质金属蛋白酶3(MMP3)/基质金属蛋白酶抑制剂2(TIMP2)比值、B细胞淋巴瘤因子(Bcl-2)等蛋白在各组大鼠心肌组织的表达变化情况。结果与对照组比较,心梗组大鼠心肌胶原容积分数增加(P<0.05),心肌细胞凋亡率升高(P<0.05),心肌组织YAP1、TAZ、MST2、Bax、Caspase-3蛋白表达及MMP3/TIMP2比值增加(均P<0.05),而Bcl-2蛋白表达降低(P<0.05);心梗十硫化氢组较心梗组大鼠心肌组织中胶原容积分数减少(P<0.05),心肌细胞凋亡率降低(P<0.05)心肌组织YAP1(2.406±0.024比2.830±0.063)、TAZ(0.964±0.090比1.329±0.018)、MST2(0.780±0.082比1.788±0.097)、Bax(1.500±0.008比0.613±0.003)、Caspase-3(0.620±0.024比0.780±0.012)蛋白表达及MMP3/TIMP2比值降低(均P<0.05),而Bcl-2蛋白表达升高(P<0.05)。结论硫化氢可改善心梗后心肌纤维化,该作用与抑制YAP1/TAZ信号通路表达和减少心肌细胞凋亡有关。Objective To investigate the effects of exogenous hydrogen sulfide on myocardial fibrosis and apoptosis in rats after myocardial infarction and the underlying mechanisms.Methods Forty-three Sprague Dawley(SD)rats were divided into 4 groups according to the random number table method:a control group(n=12),a myocardial infarction group(MI group,n=13),an hydrogen sulfide(H,S)group(n=6)and an MI+H,S group(n=12).The rat model of acute myocardial infarction was established by intraperitoneal injections of isoproterenol(50 mg/kg,once a day,for 2 days).Electrocardiogram and troponin changes were recorded 48 h after the last drug administration to determine whether the rat model was successfully constructed.After successful establishment of the model,rats in the MI group and the MI+H2S group were intraperitoneally injected with sodium hydrosulfide(56μmol/kg,once a day,for 6 weeks).6 weeks later,echocardiogram and Masson's trichrome staining were performed to assess changes in cardiac function and collagen volume fraction in each group.Terminal deoxynucleotidyl transferase(TdT)dUTP nick end labeling(TUNEL)was used to detect the myocardial apoptosis rate in each group,and Western-blot was used to detect protein expression of Yes-related protein 1(YAP1),WW domain containing transcriptional regulatorl(TAZ),mammalian Ste20-like kinase 2(MST2),Bcl-2-associated X protein(Bax),cysteine protease 3(caspase-3),the ratio of matrix metalloproteinase 3(MMP3)/matrix metalloproteinase inhibitor 2(TIMP2),and B-cell lymphoma factor(Bcl-2).Results Compared with the control group,myocardial collagen volume fraction was increased(P<0.05),the myocardial cell apoptosis rate was increased(P<0.05),and myocardial YAP1,TAZ,MST2,Bax,caspase-3 protein expression and MMP3/TIMP2 ratio were increased in the MI group(all P<0.05),while the expression of Bcl-2 protein was decreased(P<0.05).Compared with the MI group,collagen volume fraction and the cardiomyocyte apoptosis rate were significantly decreased in the MI+H2S group(P<0.05).Also,protein expressio

关 键 词：硫化氢 心肌梗死 纤维化 细胞凋亡 YAP1/TAZ信号通路 

分 类 号：R542.22[医药卫生—心血管疾病] R-332[医药卫生—内科学]