基于自噬研究穴位埋线减缓再灌注心肌无复流现象的机制

According to documents report that cell autophagy take part in resisting inflam、cardiovascular and cerebrovascular diseases 、 tumor and so on, protecting myocardium function. Our prophase research finds that after myocardial ischemia reperfusion the expression of RyR2 and SERCA that with ERS all be depressed. And through acupuncture up-regulate both of them to lessen ischemia reperfusion injury and to contract area of no-reflow. The mechanism hasn't been clarity. Accordingly, we presume that acupuncture maybe through inducing autophagy to protect ischemia myocardium and relieve area of no-reflow. The project choose Zucker Diabetic Fatty Rats and will duplicate the model of myocardial ischemia reperfusion injury, use molecular biology and electron microscope and so on; confirm the form of autophagome of Zucker Diabetic Fatty Rats' myocardial infarction reperfusion by catgut implantation at acupoint, expression of autophagy marker protein as well as influence of no-reflow. Some methods such as percutaneous coronary intervention to cure acute myocardial infarction will cause myocardial reperfusion injury and show no-reflow phenomenon frequently. So, from new view angle, the topic will command to interpret the mechanism of catgut implantation at acupoint protect cardiac muscle and relieve area of no-reflow. And provide the method of innovative non-drug therapy for preventive treatment of no-reflow, clear and definite a function target of protect myocardium by therapy of catgut implantation at acupoint.

细胞自噬参与对抗炎症、心脑血管疾病、肿瘤等，具有心肌保护作用。我们前期研究发现心肌缺血再灌注后与内质网应激相关的RyR2及SERCA均下降，且针灸可通过上调二者减轻缺血再灌注损伤并缩小无复流区域面积，但其机制尚不清晰。我们推测针刺可能通过内质网应激诱导自噬而发挥其心肌保护和减缓无复流现象的作用。本项目选择ZDF大鼠通过复制心肌缺血再灌注模型，采用分子生物学、电镜等技术，旨在明确穴位埋线对ZDF大鼠心肌梗死再灌注自噬的发生，其标志性蛋白的表达及无复流的影响。故本项目将从新的视角阐明穴位埋线保护心肌，减少无复流发生的机制，为防治无复流提供创新性非药物治疗方法、并明确埋线疗法心肌保护疗效的作用靶点。

自噬是把“双刃剑”，适度自噬有利于细胞的存活。细胞自噬参与心肌缺血再灌注全过程，且具有保护心肌的作用，但信号转导途径尚不清晰。申请人前期研究发现，针灸可以上调与内质网应激相关的RyR2及SERCA的表达，减轻再灌注损伤及缩小无复流区域面积。本项目在前期工作基础上，通过western blot、PCR等寻找在内质网应激、凋亡及自噬之间的互作靶蛋白，结合分子生物学、蛋白质生物化学、细胞生物学、电镜等方法和技术，分析研究穴位埋线参与调控保护缺血再灌注损伤心肌的作用，深入探讨靶蛋白在信号转导过程中的分子机制。此研究为进一步阐明穴位埋线保护再灌注损伤心肌，减轻无复流现象的机制，为防治无复流提供创新性非药物治疗方法提供理论依据。