基于PI3K/Akt/GSK-3β信号通路探讨稳心丹对MIRI大鼠心肌保护作用的机制研究

Action Mechanism of Wenxin Dan on Myocardial Protection in MIRI Rats Based on PI3K/Akt/GSK-3βSignaling Pathway·

目的:探讨稳心丹对心肌缺血再灌注损伤大鼠心肌保护作用及机制研究。方法:将SPF级SD大鼠,随机分为假手术组、模型组、稳心丹组、LY294002组,每组6只。采取左冠状动脉前降支先结扎30 min,再灌注60 min。酶联免疫法检测血清中CK、LDH含量;TTC染色法计算大鼠心肌梗死面积;TUNEL法检测细胞凋亡指数;Western blot法检测p-Akt、p-GSK-3β(Ser9)蛋白表达。结果:与假手术组比较,模型组CK与LDH含量显著增高、细胞凋亡指数明显升高、p-Akt及p-GSK-3β(Ser9)水平明显降低(P<0.01)、心肌梗死面积增大(P<0.05);与模型组比较,稳心丹组CK与LDH含量显著降低、心肌梗死面积明显减少、细胞凋亡指数明显降低、p-Akt及p-GSK-3β(Ser9)蛋白水平明显升高(P<0.01);与稳心丹组比较,LY294002组CK与LDH含量明显升高、心肌梗死面积明显增多、细胞凋亡指数明显升高、p-Akt及p-GSK-3β(Ser9)蛋白水平明显降低(P<0.01)。结论:稳心丹对心肌具有保护作用,其机制与激活PI3K/Akt/GSK-3β信号通路、抑制心肌细胞凋亡、减轻心肌组织损伤有关。

Objective:To explore the protective effect and action mechanism of Wenxin Dan on myocardial ischemia-reperfusion injury in rats.Methods:The SD rats were randomly divided into the sham-operation group,the model group,Wenxin Dan group and LY294002 group,with 6 rats in each group.The left anterior descending coronary artery was ligated for 30 minutes,then reperfusion was carried out for 60 minutes.ELISA method was used to to detect serum levels of CK and LDH;TTC staining method was used to calculate the area of myocardial infarction in rats;TUNEL method was applied to detect the apoptosis index;Western blot method was applied to detect protein expressions of p-Akt and p-GSK-3β(Ser9).Results:Compared to the sham operation group,the levels of CK and LDH were significantly increased,also apoptosis index was significantly increased,but the protein expressions of p-Akt and p-GSK-3β(Ser9)were significantly decreased in the model group(P<0.01);the area of myocardial infarction significantly increased in the model group(P<0.05).Compared to the model group,the contents of CK and LDH were significantly decreased,the area of myocardial infarction was significantly reduced,the apoptosis index was significantly decreased,and the protein expressions of p-Akt and p-GSK-3β(Ser9)were significantly increased in Wenxin Dan group(P<0.01).Compared to Wenxin Dan group,the levels of CK and LDH were significantly increased,the area of myocardial infarction was significantly increased,also the apoptosis index was significantly increased,but the protein expressions of p-Akt and p-GSK-3β(Ser9)were significantly reduced in LY294002 group(P<0.01).Conclusion:Wenxin Dan make a protective effect on the myocardium,and its mechanism is related to the inhibition of myocardial cell apoptosis and reduction of myocardial tissue damage by activating the PI3 K/Akt/GSK-3βsignal pathway.