HDACs-GSK3β信号通路在2型糖尿病削弱阿片类药物后处理心肌缺血再灌注损伤保护作用中的机制研究

Opioid postconditioning has a protective effect to nondiabetic myocardial ischemia- reperfusion injury (IRI), but our study showed that this effect was not so apparent during Cardiopulmonary bypass(CPB) in the cardiovascular patients with type 2 diabetes (T2D). Although limited number of basic researches revealed that the T2D myocardia might regulate insulin resistance through the HDACs-GSK3β signaling pathway, thereby put the myocardial protective effect under control, The mechanism of which still need further elucidation. In our study we would take sufentanil as an example to investigate the change in the opoid postconditioning effect on myocardia and the mechanism of which in diabetic rats with cardiovascular disease (T2D-CVD). That would include: 1.Using pharmacological methods to interfere in the HDACs-GSK3β signaling pathway, and observe the induced myocardial protection; 2. Taking advantage of a combination of a morphological method of immunofluorescence / immunoelectron microscopy and the molecular biology technique of DNA Co-Immunoprecipitation, to reveal the molecular and cellular mechanisms of myocardial IRI through opioid receptor / HDACs - GSK3β signaling pathway. The destination is to find better myocardial protection targets, so as to lay theoretical foundation for developing new myocardial protective drugs, and for overcoming the difficult problem of proventing and treating myocardial IRI underlying the co-mobidity of T2D with CVD.

阿片类药物后处理对于非糖尿病心肌缺血/再灌注损伤具有保护效应。本课题组临床研究表明，体外循环心脏手术中阿片类药物后处理心肌保护作用对于合并2型糖尿病(T2D)的心血管疾病(CVD)患者并不明显。有限的基础研究提示，T2D心肌可能通过HDACs-GSK3β途径调节胰岛素抵抗、控制内源性阿片类物质与受体的表达。本项目拟以舒芬太尼为代表，研究T2D-CVD大鼠阿片类药物后处理的心肌保护作用及机制：①采用药理学手段干预HDACs-GSK3β信号途径，观察其心肌保护效果；②结合免疫荧光/免疫电镜等形态学手段、染色质免疫共沉淀等分子生物学手段，揭示阿片类物质及其受体/HDACs-GSK3β信号途径调控T2D-CVD心肌缺血-再灌注损伤的细胞与分子机制。期望发现更为理想的心肌保护作用靶点，为研发心肌保护新药、解决T2D-CVD共病条件下心肌缺血-再灌注损伤之临床防治难题提供理论依据。

本项目基于既往研究发现，阿片类药物后处理对2型糖尿病(T2DM)大鼠心肌缺血再灌注损伤的保护作用并不明显，结合相关预实验结果，提出“T2DM心肌可能通过组蛋白乙酰化-GSK3β信号通路调节胰岛素抵抗、控制心肌保护效应”；以舒芬太尼为代表，研究基于HDACs-GSK3β信号途径介导的T2DM大鼠削弱阿片类药物后处理心肌保护作用的机制：①采用药理学手段干预HDACs-GSK3β信号途径，观察其心肌保护效果；②干预阿片类物质及其受体，观察其对HDACs-GSK3β信号分子表达的影响。发现HDACs的差异性表达可通过乙酰化与去乙酰化途径调节GSK3β的磷酸化修饰，调控T2D的心肌缺血-再灌注损伤、影响舒芬太尼后处理的心肌保护作用；但鉴于心肌细胞内乙酰化修饰作用(组蛋白与非组蛋白机制)的广泛性与复杂性，其确切的表观遗传修饰机理仍有待于进一步研究。该研究初步探讨了HDACs-GSK3β信号途径调控T2D-CVD心肌缺血-再灌注损伤的细胞与分子机制；为阐明T2D削弱阿片类药物心肌保护作用的确切机理提供了一定的理论依据。.受本项目资助，发表学术论文2篇，培养研究生6名(博士生1人、硕士生5人，毕业2人）；衍生出省自然科学基金等课题2项。