吡格列酮对缺血再灌注乳鼠心肌细胞凋亡的作用

Effect of pioglitazone on rat cardiomyocytes apoptosis induced by ischemia reperfusion injury

目的：研究吡格列酮对离体缺血再灌注损伤心肌细胞凋亡的影响及机制,为吡格列酮治疗心肌缺血再灌注损伤提供理论基础和实验依据.方法：体外培养Wistar乳鼠心室肌细胞,建立心肌细胞缺氧复氧模型.应用免疫荧光显色检测心肌细胞Caspase 3表达变化,免疫印迹法检测各组总Akt、 p-Akt蛋白的表达变化.结果：吡格列酮使离体培养的缺血再灌注损伤心肌细胞Caspase 3阳性细胞减少,p-Akt活性增加,这一作用可被Akt抑制剂Ly294002阻断.结论：吡格列酮激活PI3K/Akt通路,减轻离体缺血再灌注心肌细胞凋亡.

Objective.. To study the effect of pioglitazone on in vitro eardiomyocytes apoptosis induced by ischemia reperfusion injury and to provide rationale basis and experiment evidence for the treatment of ischemia reperfusion injury of myocardial cells. Methods： Newborn Wistar rats hearts were isolated and cardiomyocytes were separated by trypsin digestion. Hypoxia reoxygenation injury was established by 3 hours of hypoxia and subsequently 3 hours of reoxygenation. The cardiomyocytes were cultivated in vitro and alloted to 4 groups： a normal control group, an ischemia/reperfusion injury group, pioglitazone preconditioning group, and Ly294002 （inhibitor of Akt） group. Expressions of Caspase 3 were detected by immunofluorescence technique. Western blotting was used to analyse the expression of total Akt and phosphorylated Akt in all groups. Re- sults： The expression of Caspase 3 was decreased and the activity of Akt was increased on the contrary. There were statistically significant differences. Those changes were inhibited by Ly294002. Conclusion： Pioglitazone may protect cardiomyocytes from ischemia/reperfusion-induced apoptosis through PI3K/Akt signaling pathway.