DNA加双氧酶TET1在高压诱导心肌纤维化中的作用研究  

Role of DNA dioxygenase TET1 in cardiac fibrosis induced by high pressure

作　　者：李路安 吴清蕊 李倩 饶芳 邝素娟 杨慧 张黔桓[2] 张斌 LI Lu-an;WU Qing-rui;LI Qian;RAO Fang;KUANG Su-juan;YANG Hui;ZHANG Qian-huan;ZHANG Bin(School of Medicine,South China University of Technology,Guangzhou 510006,China;Dept of Cardiology,Guangdong Institute of Cardiovascular Diseases,Guangzhou 510080,China;Key Laboratory of Clinical Pharmacology,Medical Research Dept,Guangdong People's Hospital(Guangdong Academy of Medical Sciences),Guangzhou 510080,China)

机构地区：[1]华南理工大学医学院,广东广州510006 [2]广东省心血管病研究所心血管内科,广东广州510080 [3]广东省人民医院(广东省医学科学院)医学研究部临床药理重点实验室,广东广州510080

出　　处：《中国药理学通报》2021年第7期922-928,共7页Chinese Pharmacological Bulletin

基　　金：国家自然科学基金资助项目(No.81470505)。

摘　　要：目的探讨TET1在高压诱导心肌纤维化中的表达及作用机制。方法选取Wistar大鼠和自发性高血压大鼠(SHR),Western blot检测心室肌组织中TET1、TGF-β、COL-1和COL-3表达,HE及Masson染色检测心肌病理学改变。选取原代SD乳大鼠心肌成纤维细胞(NRCFs),分别给予0、120及180 mmHg的压力干预24 h,诱导心肌纤维化模型;免疫荧光检测5-hmC的变化;qRT-PCR检测TGF-β启动子区域5-hmC与5-mC的变化;Western blot检测TET1、TGF-β、COL-1和COL-3的表达。结果SHR较Wistar大鼠血压升高,心肌胶原纤维增多,TET1、TGF-β、COL-1和COL-3表达明显增加。与0 mmHg相比,120 mmHg和180 mmHg促进NRCFs中TET1、5-hmC、TGF-β、COL-1和COL-3的增加;180 mmHg压力下敲低TET1明显缓解了5-hmC、TGF-β、COL-1和COL-3的增加,并降低了TGF-β启动子区域5-hmC的水平、且增加其5-mC和5-hmC的总水平。结论高压负荷诱导的心肌纤维化,其机制可能与TET1促进TGF-β启动子去甲基化并促使TGF-β表达增加有关。Aim To investigate the mechanism of TET1 in cardiac fibrosis induced by high pressure.Methods Wistar rats and spontaneous hypertension rats(SHR)were selected to detected the expression of TET1,TGF-β,COL-1 and COL-3 in myocardium by Western blot;HE and Masson staining were used to detect myocardial pathological changes.Neonatal rat cardiac fibroblasts(NRCFs)were isolated from the ventricles of neonatal Sprague-Dawley rats and stimulated by 0 mmHg,120 mmHg and 180 mmHg high pressure.Immunofluorescence was used to detect the changes of 5-hmC in the NRCFs.The changes of 5-hmC and 5-mC in TGF-βpromoter region were detected by qRT-PCR.The expressions of TET1,TGF-β,COL-1 and COL-3 were detected by Western blot.Results Compared with Wistar rats,SHR showed increased blood pressure,increased fibrous collagen in ventricular tissues,and significantly increased expressions of TET1,TGF-β,COL-1 and COL-3.Compared with the 0 mmHg group,120 mmHg and 180 mmHg group significantly induced the increase of TET1,5-hmC,TGF-β,COL-1 and COL-3.TET1 knockdown significantly reduced the increase of 5-hmC,TGF-β,COL-1 and COL-3 under 180 mmHg pressure.Besides,knockdown TET1 significantly reduced the level of 5-hmC and increased the level of 5-mC and 5-hmC in the TGF-βpromoter region.Conclusions High pressure induced cardiac fibrosis is associated with the promotion of TGF-βpromoter demethylation and the increased of TGF-βexpression by TET1.

关 键 词：TET1 TGF-Β 5-hmC 高压 心肌纤维化 去甲基化 

分 类 号：R-332[医药卫生] R322.11