吡格列酮对血管内皮细胞及JNK信号通路的影响

The effects of pioglitazone on vascular endothelial cell and c-Jun N-terminal kinase signalling pathway

目的探讨吡格列酮(PIO)对血管内皮细胞的保护作用及其可能机制。方法用四唑盐比色法(MTT法)检测PIO对高糖损伤血管内皮细胞的最佳保护浓度;按不同干预条件,将血管内皮细胞分为7组:正常培养组[A组,葡萄糖(Glu)5.5 mmol/L],高糖培养组(B组,Glu 37.5 mmol/L),C、D、E组分别为不同浓度PIO+高糖培养,F、G组为不同浓度SP600125+PIO+高糖培养。Western blot法检测各组c-Jun氨基末端激酶(JNK)、磷酸化c-Jun氨基末端激酶(p-JNK)蛋白表达。结果 PIO在1×10-8~1×10-4mmol/L时对高糖损伤的血管内皮细胞有保护作用;与B组相比,C^E组p-JNK表达降低,差异有统计学意义(P<0.05)。加用JNK抑制剂SP600125后,p-JNK表达进一步下降,差异有统计学意义(P<0.05)。结论 PIO可提高高糖损伤的血管内皮细胞存活率,降低JNK及p-JNK蛋白表达,其机制可能是PIO具有抑制JNK信号通路活性的作用。

Objective To investigate the protective effect and possible mechanism of pioglitazone（PIO） on vascular endothelial cell.Methods The optimum concentration of PIO played a protective role on cells which was assessed by MTT assay.The vascular endothelial cells were randomly divided into 7 groups,normal culture group [group A,glucose（Glu） 5.5mmol/L],cultured in high glucose group（group B,Glu 37.5 mmol/L）,the cells in group C,D and E were cultured in PIO（with different concentration）＋ high glucose,and group F and G were cultured in SP600125（with different concentration）＋PIO＋high glucose.The protein expression of c-Jun N-terminal kinase（JNK） and JNK phosphorylation（p-JNK） in every group were detected by Western blot.Results PIO had a protective effect on high glucose injured vascular endothelial cell at the concentrations of 1×10-8~1×10-4 mmol/L.Compared to group B,the level of p-JNK in group C^E decreased obviously,there was significant difference（P0.05）.Compared to the PIO group,JNK inhibitor SP600125 could decline the level of p-JNK further,the difference was significant（P0.05）.Conclusion PIO can improve the survival rate of vascular endothelial cell with high glucose damage,decrease the expression of JNK and p-JNK.The effect of PIO on vascular endothelial cell can be enhanced by the JNK inhibitor SP600125,and the protective effect of PIO may be achieved by affecting the JNK pathway.