吡格列酮对缺氧复氧大鼠心肌细胞凋亡及低氧诱导因子1α的影响

The Effect of Pioglitazone on the Hypoxia-reoxygenation Induced Cardiac Myocytes Apoptosis and the Expression of HIF-1α

目的观察吡格列酮对缺氧复氧大鼠心肌细胞凋亡的保护作用,并探讨吡格列酮对低氧诱导因子-1α(HIF-1α)表达的影响。方法原代培养的SD乳鼠心肌细胞随机分为4组:溶媒组、缺氧复氧+溶媒组、缺氧复氧+吡格列酮0.1μM组、缺氧复氧+吡格列酮1μM组,建立缺氧复氧模型,利用Annexin-v与PI双染法及流式细胞仪检测心肌细胞凋亡,RT-PCR及Western blot方法检测HIF-1α mRNA及蛋白表达的变化。结果缺氧复氧后,溶媒组心肌细胞发生明显凋亡,吡格列酮以剂量依赖方式减少心肌细胞凋亡(P<0.05);缺氧复氧组HIF-1α表达上调,吡格列酮各组促进其进一步上调(P<0.05),与剂量无关。结论吡格列酮对缺氧复氧大鼠心肌细胞凋亡具有保护作用,其机制可能与上调HIF-1α表达有关。

Objective To investigate the effect of pioglitazone on the hypoxia - reoxygenation induced cardiac myocytes apoptosis and the expression of HIF -1α. Methods Cultured cardiomyocytes of neonatal Sprague - Dawley rats were divided into four groups： vehicle, hypoxia/reoxygenation ＋ vehicle, hypoxia/reoxygenation ＋ pioglitazone 0.1μM, hypoxia/ reoxygenation ＋ pioglitazone 1μM. We measured the apoptosis rate of cardiomyocytes after hypoxia/reoxygenation. RT - PCR and Western blot analysis were performed to detect expressions of HIF -1α mRNA and protein expression. Results Apoptosis index in hypoxia/reoxygenation ＋ vehicle group significandy increased than in the vehicle group. Pioglitazone decreased apoptosis index in dose - dependent manner （ P 〈 0. 05 ）. Hypoxia/reoxygenation induced upregulation of HIF - Ict both in mRNA and protein level. Pioglitazone further increased the expression of HIF -1α （P 〈0. 05 ）. Conclusion Pioglitazone can protect the cardiomyocyte from I/R injury by reducing cardiomyocyte apoptosis. The protective effect is likely to occur by regulation of HIF -1α expression.