心肌缺血再灌注(ischemia/reperfusion, I/R)损伤是体外循环术后常见的心脏损害，如何减轻I/R对心脏的损伤一直是医学界研究的热点。自噬是细胞内普遍存在的一种降解机制，对维持细胞稳态有重要意义。研究表明心肌细胞自噬在I/R状态下激活并参与心肌损伤，但其功能与机制尚未阐明。长链非编码RNA作为一种新的转录本，调节各种病理生理过程。我们前期发现HIF1A-AS1在I/R损伤的心肌组织中表达异常，抑制其水平降低缺/复氧诱导的心肌细胞损伤，并且对自噬标记蛋白beclin 1的表达具有调节作用。基于此提出假设：长链非编码RNA HIF1A-AS1通过调节心肌细胞自噬水平参与心肌I/R损伤。我们拟利用基因干预技术、电镜技术和基因芯片等研究HIF1A-AS1调控自噬的机制，阐明心肌I/R损伤中HIF1A-AS1及自噬的功能。研究成果将为缺血性心脏病的治疗及预后提供新的方法和研究方向。

Myocardial ischemia reperfusion (I/R) injury is common for cardiac damage after cardiopulmonary bypass operation, and how to reduce I/R damage of heart is always a hotspot in medical research. Autophagy is the universal mechanism of degradation, playing an important role on homeostasis maintain. Recent research had showed that myocardial autophagy activated under I/R state, and participate in myocardial injury, but the function and the mechanism has not been clarified. As a new transcripts, long noncoding RNA was proved to regulate various pathological and physiological process. Our previous study had found the abnormal expression of HIF1A-AS1 in I/R damage cardiac tissue, and inhibition of HIF1A-AS1 reduce the myocardial cell injury induced by hypoxia/reoxygenation, which might by regulating the autophagy marker protein Beclin 1 expression. Therefore, hypothesis is proposed: The long non-coding RNA HIF1A-AS1 regulated autophagy of cardiomyocytes in myocardial ischemia reperfusion injury. We intended to study the mechanism of HIF1A-AS regulating autophagy by gene intervention technique, electron microscopy and gene chip assay, and to clarify the function of HIF1A-AS1 and autophagy in myocardial I/R injury. The research findings would provide a new research method and direction for the treatment and prognosis of ischemic heart diseases.

心肌缺血再灌注(ischemia/reperfusion, I/R)损伤是体外循环术后常见的心脏损害，如何减轻I/R对心脏的损伤一直是医学界研究的热点。自噬是细胞内普遍存在的一种降解机制，对维持细胞稳态有重要意义。研究表明心肌细胞自噬在I/R状态下激活并参与心肌损伤，但其功能与机制尚未阐明。长链非编码RNA作为一种新的转录本，调节各种病理生理过程。本项目主要通过构建心肌细胞缺/复氧损伤模型，检测HIF1A-AS1与细胞自噬标记蛋白的表达水平，明确HIF1A-AS1与心肌细胞自噬以及损伤的相关性。同时通过心肌细胞中抑制HIF1A-AS1水平，检测了缺/复氧诱导后，细胞自噬活力的变化，心肌细胞损伤的变化，明确HIF1A-AS1对心肌细胞自噬的调控作用。定量PCR结果显示HIF1A-AS1在心肌细胞缺/复氧刺激后表达明显升高，western blot结果显示细胞自噬标志蛋白Beclin 1的表达也明显升高，HIF1A-AS1水平与自噬成正相关。心肌细胞中抑制HIF1A-AS1后，经缺/复氧刺激，Beclin 1的表达明显低于对照组，自噬活性显著降低，培养液中LDH含量明显降低。结果说明抑制HIF1A-AS1对心肌细胞缺复氧损伤有保护作用，将为缺血性心脏病的治疗及预后提供新的方法和研究方向。