蛋白激酶D1对心肌梗死大鼠心肌组织炎症和凋亡的影响

Effect of protein kinase D1 on myocardial inflammation and apoptosis in rats with myocardial infarction

目的探讨蛋白激酶D1(PKD1)对心肌梗死后心肌组织的炎症和凋亡的影响,并分析其分子机制。方法45只♂Wistar大鼠随机分为假手术(Sham)组、模型组和PKD1组,每组15只。模型组和PKD1组结扎左前降支冠状动脉复制心肌梗死模型,Sham组仅手术而不结扎血管。评估血流动力学参数,HE染色法分析心肌组织形态学变化,TUNEL、免疫组化和免疫印迹法分析心肌细胞凋亡变化,免疫印迹和实时定量PCR(qPCR)法分析心肌组织炎症反应调控蛋白和炎症因子表达变化。结果与模型组相比,PKD1可改善心梗大鼠的血流动力学指标,减轻心肌梗死引起的组织损伤,抑制心肌细胞凋亡,上调Bcl-2表达,下调caspase-3、Bax、TLR4、TN-C、NF-κBp50和NF-κBp65蛋白的表达,同时下调IL-1、NF-κBp50、NF-κBp65mRNA的表达,差异均有统计学意义(P<0.01)。结论PKD1可能有对抗心肌梗死损伤引发的炎症和细胞凋亡作用。

Aim To investigate the effect of protein kinase D1(PKD1) on myocardial inflammation and apoptosis after myocardial infarction and to analyze its molecular mechanism.Methods Forty-five male Wistar rats were randomly divided into three groups:sham-operated group,model group and PKD1 group.Rat models of myocardial infarction were reproduced by classical left coronary artery ligation in model group and PKD1 group,while rats in sham group were operated without ligation of coronary artery.The parameters of hemodynamics in rats were assessed,the morphological changes of myocardial tissue were analyzed by HE staining,the apoptotic changes of myocardial cells were analyzed by TUNEL staining,immunohistochemical staining and Western blot,while the changes of myocardial inflammation were analyzed by immunoblotting and real-time quantitative PCR.Results Compared with model group,PKD1 could improve the hemodynamic indexes in rats with myocardial infarction,reduce the injury caused by myocardial infarction,inhibit apoptosis in myocardial tissue,up-regulate the protein expression of Bcl-2,down-regulate the protein expression of caspase-3,Bax,TLR4,TN-C,NF-κB p50,NF-κB p65 and decrease the mRNA expression of IL-1,NF-κB p50 and NF-κB p65,and all the differences were statistically significant( P <0.01).Conclusion PKD1 might have the biological function of inhibiory effect on inflammation and apoptosis induced by myocardial infarction injury.