基于ERK1/2/GATA4通路对“气虚血瘀”致心肌缺血再灌注损伤的作用及益气活血中药干预机制研究

心肌缺血再灌注损伤(MIRI),严重危险人类健康.“气虚血瘀,脉络受阻”是MIRI的重要病机。我们前期研究发现益气活血中药通冠胶囊能改善心肌缺血再灌注损伤，但其确切机制尚未阐明。课题组前期研究发现, ERK1/2/GATA4通路在再灌注损伤中起到重要作用。我们推测“气虚血瘀”，过度激活ERK1/2/GATA4通路；通冠胶囊通过抑制ERK1/2/GATA4通路，减少心肌细胞凋亡，从而改善MIRI。为证实该假说，本研究以18月龄老年小鼠冠脉紧松的方法构建气虚血瘀型MIRI模型，通过心肌切片染色明确心肌梗死，心脏超声明确心功能，Tunnel法检测心肌细胞凋亡，免疫印迹检测ERK1/2/GATA4蛋白表达，EMSA、Chip检测GATA4与凋亡基因Bcl-2启动子结合情况，并且观察通冠胶囊对气虚血瘀型MIRI的作用机制。本研究的开展为通冠胶囊治疗MIRI的临床应用提供新的作用靶点和试验依据。

Myocardial ischemia and reperfusion injury (MIRI), is a serious risk factor of human health. "Qi deficiency and blood stasis, the context is blocked" is an important pathogenesis of MIRI. Our previous study showed that Yiqi Huoxue drug,Tongguan capsule can reduce MIRI, but the exact mechanism has not been elucidated. In the previous studies, the ERK1/2/GATA4 pathway played an important role in reperfusion injury. We hypothesized that deficiency of Qi and blood stasis, excessive activation of ERK1/2/GATA4 pathway; Tongguan Capsule can inhibit the pathway of ERK1/2/GATA4, reducing myocardial cell apoptosis, so as to improve MIRI. . To prove the hypothesis, we are going to construct the MIRI model of Qi deficiency and blood stasis type by tightening and loosening the coronary artery of 18 months old mice, observing infarction size by Evans blue/TTC staining, calculating heart function by echocardiography, watching myocardial cell apoptosis by tunnel method, ensuring ERK1/2/GATA4 protein expression by Western blotting, detecting the combination of GATA4 with Bcl-2 promoter binding site, and revealing the mechanisms of Tongguan Capsule on MIRI model of Qi deficiency and blood stasis type. This study was undertaken to provide targets and experimental basis for new clinical application of Tongguan Capsule in the treatment of MIRI.