GSK-3β对mPTP的调控在活性成分保护HPM辐射致大鼠心脏损伤中的作用研究

Previous studies have shown that microwave radiation can cause multiple organ damages in human body, and the heart is one of the organs that are most susceptible to the microwave-induced injury. Recently, a great deal of attention has been paid on the research of high power microwave-induced injury and its mechanism both at home and abroad. However, there is not effective drugs that could protest against HPM-induced injury yet. The present study was based on our studies previously carried out. First of all, animal models of the microwave-induced myocardial injury have been established and received mediation, and the changes of the cardiac structure and function have been observed; Secondly, the changes of the important moleculars indluding GSK - 3 beta, p-GSK-3β, VDAC, and p-VDAC in GSK-3β/mPTP pathway have been detected ; Finally, an in vitro model of microwave radiation induced myocardial cell damage has been established and received mediation, to explore the pharmacological mechanism of the modulation of mPTP by GSK-3β in heart protection. This study was to explore the protection of active ingredients against HPM-induced myocardial injury in rats, and to explore the regulatory mechanism based on the modulation of mPTP by GSK-3β, and then to develop effective drugs against HPM-induced myocardial injury and to explore the effective targets, and to provide important experimental basis on the development of drugs protecting against microwave-induced injury.

研究表明，微波辐射可造成人体多器官损伤，心脏是其致伤的敏感器官之一。近年来，对HPM辐射损伤及其机制的研究在国内外受到普遍重视，但有效的防护药物尚缺乏。为此，本研究在既往研究基础上。首先，建立微波辐射心脏损伤动物模型并给予药物治疗，观察心脏结构及功能变化；其次，检测GSK-3β/mPTP通路中GSK-3β、p-GSK-3β、VDAC、及p-VDAC等重要分子的变化情况；最后，建立微波辐射致体外心肌细胞损伤模型并给予药物治疗，探讨GSK-3β调控mPTP的心脏保护作用的药理机制。本研究拟在探讨抗辐灵及其活性成分对HPM辐射致大鼠心脏损伤保护作用的基础上，以GSK-3β对mPTP的调控为切入点，寻找活性成分对HPM辐射致心脏损伤保护作用的调控机制，寻找微波辐射致心脏损伤有效的防治药物及有效作用靶点，为微波损伤防治药物的研发提供重要的实验依据。

摘要: 目的 探讨GSK-3β对mPTP的调控在活性成分保护HPM辐射致大鼠心脏损伤中的作用研究。 方法 采用30mW/cm2的微波辐射H9c2心肌细胞，检测ROS含量和心肌酶活性，筛选黄芪总苷、赤芍总苷和丹参总酮中药效最佳活性；30mW/cm2的HPM辐射Wistar大鼠，给予三个剂量黄芪总苷治疗，通过对大鼠心电图、血清中心肌酶及Ca2+、心脏组织学结构及超微结构观察，明确黄芪总苷对HPM辐射致心脏损伤的保护作用和最佳有效剂量；建立细胞损伤模型，黄芪总苷，辐射前给予GSK-3β特异性的抑制剂进行干预，检测GSK-3β调控mPTP相关蛋白的表达、mPTP开放及对线粒体功能的影响。探索黄芪总苷对HPM辐射所致大鼠心脏损伤可能的保护机制，寻找黄芪总苷保护HPM辐射心脏损伤有效作用靶点。结果 30mW/cm2微波辐射可引起H9c2心肌细胞CK、LDH活性及ROS含量升高，黄芪总苷、赤芍总苷及丹参总酮对上述指标改变的改善效果良好，其中黄芪总苷改善效果最佳，最佳有效剂量为3mg/l；30mW/cm2微波辐射可致大鼠心脏功能损伤，心脏组织结构及超微结构损伤，80mg/kg/d黄芪总苷药物治疗改善效果最佳；30mW/cm2HPM辐射致细胞活力降低，mPTP开放、膜电位降低，GSK-3β磷酸化及VDAC蛋白表达降低，细胞中ROS含量升高及ATP含量减少，线粒体Ca2+含量降低。1-Azakenpaullone抑制剂组及黄芪总苷药物组可抑制上述各指标的改变。结论：（1）微波辐射引起H9c2心肌细胞ROS含量及心肌酶活性升高，3mg/l黄芪总苷改善效果最佳（2）80mg/kg/d黄芪总苷对微波辐射所致的大鼠心脏功能紊乱和组织结构损伤具有良好的保护作用（3）微波辐射可致H9c2心肌细胞活力降低，线粒体功能异常，GSK-3β、VDAC蛋白表达异常，黄芪总苷改善效果佳；（4）GSK-3β是HPM辐射心肌细胞损伤重要靶分子，黄芪总苷对HPM致心肌细胞损伤的保护机制是通过磷酸化GSK-3β，抑制其活性、促进VDAC蛋白表达，从而抑制mPTP的开放、改善线粒体功能来实现。