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长链非编码RNA HIF1A-AS1对大鼠心肌缺血再灌注损伤的调控作用
张冠鑫1,丛滨海2,张加俊1,王崇1,袁扬1,王国坤1,韩林1,徐志云1*
0
(1. 第二军医大学长海医院胸心外科, 上海 200433;
2. 第二军医大学基础医学部生理学教研室, 上海 200433
*通信作者)
摘要:
目的 研究长链非编码RNA HIF1A-AS1对大鼠缺血再灌注损伤的调控作用,并初步探索其机制。方法 构建大鼠心肌缺血再灌注损伤和心肌细胞缺氧/复氧损伤模型,利用实时定量PCR检测HIF1A-AS1的表达。采用siRNA抑制心肌细胞HIF1A-AS1的表达并制备缺氧/复氧损伤模型,用MTT法检测心肌细胞生长活力,ELISA法检测培养液中乳酸脱氢酶的水平,蛋白质印迹法检测自噬相关蛋白Beclin-1的表达变化。结果 HIF1A-AS1在缺血再灌注大鼠心肌组织和缺氧/复氧损伤心肌细胞中表达上调。抑制HIF1A-AS1表达可保护心肌细胞,逆转缺氧/复氧刺激导致的心肌细胞生长活力降低、乳酸脱氢酶分泌水平增高、自噬标志蛋白Beclin-1表达增高现象。结论 抑制长链非编码RNA HIF1A-AS1,可能通过抑制心肌细胞的过度自噬抵抗缺血再灌注诱导的心肌损伤。
关键词:  长链非编码RNA  HIF1A-AS1  自噬  心脏肌细胞  心肌再灌注损伤
DOI:10.3724/SP.J.1008.2015.00131
投稿时间:2014-12-18修订日期:2015-01-12
基金项目:上海市自然科学基金 (15ZR1413400).
Regulatory effects of long non-coding RNA HIF1A-AS1 on ischemic myocardial reperfusion injury in rats
ZHANG Guan-xin1,CONG Bin-hai2,ZHANG Jia-jun1,WANG Chong1,YUAN Yang1,WANG Guo-kun1,HAN Lin1,XU Zhi-yun1*
(1. Department of Cardiothoracic Surgery, Changhai Hospital, Second Military Medical University, Shanghai 200433, China;
2. Department of Physiology, College of Basic Medical Sciences, Second Military Medical University, Shanghai 200433, China
*Corresponding author)
Abstract:
Objective To study the regulatory effects of long non-coding RNA HIF1A-AS1 on the myocardial ischemia reperfusion (I/R) injury and the related mechanism. Methods Myocardial I/R injury model was established with SD rats, and hypoxia reoxygenation (H/R) model was established with rat cardiac myocytes. si-HIF1A-AS1 was used to inhibit HIF1A-AS1 expression in the cardiac myoctyes. Then the mRNA expression of HIF1A-AS1 was detected by real-time PCR, the growth vitality of cardiac myocytes was investigated by MTT assay, the concentration of lactate dehydrogenase (LDH) in the culture media was detected by ELISA, and the autophagy-associated protein Beclin-1 expression was observed by Western blotting analysis. Results HIF1A-AS1 expression was increased in cardiac muscle of rat I/R model and rat cardiac myocytes of H/R model. Inhibition of HIF1A-AS1 by siRNA protected the cardiomyocytes against H/R injuries, reversing the decreased growth vitality of cardiac myoctyes, increased LDH level in the culture media, and increased expression of autophagy-related protein Beclin-1 induced by H/R stimulation. Conclusion Inhibition of long non-coding RNA HIF1A-AS1 might play a protective role in I/R injury of cardiac myoctyes by inhibiting the excessive autophagy of cardiomyocytes.
Key words:  long non-coding RNA  HIF1A-AS1  autophagy  cardiac myocytes  myocardial reperfusion injury