CircFoxo3调节miR-130a-5p/TEAD1轴对心力衰竭大鼠心肌细胞凋亡影响的实验研究  

An experimental study on the effect of CircFoxo3 regulating miR-130a-5p/TEAD1 axis on myocardial cell apoptosis in heart failure rats

作　　者：曹彦花 乐金海 陈新宇[1] 刘建和[1] 刘越美[1] 李俊 李苏[1] CAO Yanhua;LE Jinhai;CHEN Xinyu;LIU Jianhe;LIU Yuemei;LI Jun;LI Su(Department of Cardiovascular Medicine,the First Hospital of Hunan University of Chinese Medicine,Changsha 410000,China;Department of Critical Care Medicine,the First Hospital of Hunan University of Chinese Medicine,Changsha 410000,China;Department of Orthopaedics and Traumatology,Changsha Hospital of Traditional Chinese Medicine,Hunan 410000,China)

机构地区：[1]湖南中医药大学第一附属医院心血管内科,长沙410000 [2]湖南中医药大学第一附属医院重症医学科,长沙410000 [3]长沙市中医医院骨伤科,湖南410000

出　　处：《中国临床新医学》2024年第1期65-73,共9页CHINESE JOURNAL OF NEW CLINICAL MEDICINE

基　　金：国家重大疑难疾病慢性心力衰竭中西医临床协作试点项目。

摘　　要：目的探讨环状RNA叉头框蛋白O3(CircFoxo3)调节miR-130a-5p/TEA域转录因子1(TEAD1)轴对心力衰竭(HF)大鼠心肌细胞凋亡的影响。方法将SD大鼠分为对照组、HF组、si-NC组、si-CircFoxo3组、agomir NC组、miR-130a-5p agomir组、si-CircFoxo3+antagomir NC组、si-CircFoxo3+miR-130a-5p antagomir组,每组18只。除对照组外,其他组大鼠均通过腹腔注射盐酸阿霉素的方法构建HF大鼠模型。建模成功后进行药物处理,每3 d给药一次,持续3周。应用反转录实时荧光定量聚合酶链式反应(qRT-PCR)法检测心肌组织中CircFoxo3、miR-130a-5p表达水平。应用彩色多普勒超声仪检测大鼠左室收缩末期内径(LVESD)、左室舒张末期内径(LVEDD)、左室射血分数(LVEF)水平。采用酶联免疫吸附试验(ELISA)法检测大鼠血清人基质裂解素2(ST_(2))、N端脑钠肽前体(NT-pro BNP)水平。应用HE染色、Masson染色检测大鼠心肌组织病理变化和心肌纤维化情况。通过TUNEL染色检测心肌细胞凋亡水平。应用Western blot法检测大鼠心肌组织TEA域转录因子1(TEAD1)、B淋巴细胞瘤-2(Bcl-2)、Bcl-2相关X蛋白(Bax)、裂解的天冬氨酸特异性半胱氨酸蛋白酶-3(cleaved caspase-3)蛋白表达水平。通过双荧光素酶报告基因实验验证CircFoxo3与miR-130a-5p、miR-130a-5p与TEAD1的关系。结果与对照组比较,HF组大鼠心肌组织病理损伤、心肌纤维化严重,LVESD、LVEDD水平上升,心肌组织CircFoxo3及TEAD1、Bax、cleaved caspase-3蛋白表达水平升高,血清ST_(2)、NT-pro BNP水平升高,心肌细胞凋亡率升高,LVEF、miR-130a-5p和Bcl-2蛋白水平降低,差异有统计学意义(P<0.05)。与HF组、si-NC组比较,si-CircFoxo3组大鼠心肌组织病理损伤减轻,LVESD、LVEDD水平降低,心肌组织CircFoxo3及TEAD1、Bax、cleaved caspase-3蛋白表达水平降低,血清ST_(2)、NT-pro BNP水平降低,心肌细胞凋亡率降低,LVEF、miR-130a-5p及Bcl-2蛋白水平升高,差异有统计学意义(P<0.05)。与HF组�Objective To explore the effect of circular RNA forkhead box protein O3(CircFoxo3)regulating micro ribonucleic acid-130a-5p(miR-130a-5p)/TEA domain transcription factor 1(TEAD1)axis on myocardial cell apoptosis in heart failure(HF)rats.Methods Sprague-Dawley(SD)rats were divided into control group,HF group,si-NC group,si-CircFoxo3 group,agomir NC group,miR-130a-5p agomir group,si-CircFoxo3+antagomir NC group and si-CircFoxo3+miR-130a-5p antagomir group,with 18 rats in each group.Except for the rats in the control group,the rats in the other groups were injected with doxorubicin hydrochloride intraperitoneally to construct HF rat models.After successful modeling,medication was administered once every 3 days for 3 weeks.Quantitative reverse transcription polymerase chain reaction(qRT-PCR)was applied to detect the expression levels of CircFoxo3 and miR-130a-5p in the myocardial tissues.Color Doppler ultrasound was applied to detect the levels of left ventricular end systolic diameter(LVESD),left ventricular end diastolic diameter(LVEDD),and left ventricular ejection fraction(LVEF)in the rats.Enzyme-linked immunosorbent assay(ELISA)method was applied to detect the levels of human stromelysin-2(ST_(2))and N-terminal pro-brain natriuretic peptide(NT-pro BNP)in the serum of the rats.Haematoxylin-eosin(HE)staining and Masson staining were used to detect the pathological changes and the myocardial fibrosis in the myocardial tissues of the rats.TUNEL staining was applied to detect the level of cardiomyocyte apoptosis.Western blot method was applied to detect the expression levels of TEAD1,B cell lymphoma-2(Bcl-2),Bcl-2-associated X protein(Bax),and cleaved aspartate-specific cysteine protease-3(cleaved caspase-3)in the myocardial tissues of the rats.Double luciferase reporter gene experiment was applied to verify the relationship between CircFoxo3 and miR-130a-5p,and between miR-130a-5p and TEAD1.Results Compared with those in the control group,the rats in the HF group had pathological injury of myocardial tissues and seve

关 键 词：环状RNA叉头框蛋白O3 miR-130a-5p TEA域转录因子1 心力衰竭 凋亡 

分 类 号：R541[医药卫生—心血管疾病]