【摘要】 目的：探讨尼莫地平对人脑血管外膜成纤维细胞活力、凋亡的影响及其对胰岛素受体底物-1(IRS-1)的调控机制。方法：体外培养人脑血管外膜成纤维细胞，分为NC组、IFN-α组和共同处理组，其中共同处理组分别采用不同浓度（50、100、200μg/ml）的尼莫地平与IFN-α共同处理人脑血管外膜成纤维细胞，分别记作IFN-α+Nimo 50μg/ml组、IFN-α+Nimo 100μg/ml组、IFN-α+Nimo 200μg/ml组；MTT检测细胞活力；流式细胞术检测细胞凋亡率；qRT-PCR与Western blot分别检测IRS-1表达；分别将pcDNA、pcDNA-IRS-1转染至细胞，分别将si-con、si-IRS-1转染至细胞随后采用尼莫地平处理细胞，采用上述方法检测细胞活力及凋亡率。结果：NC组、IFN-α组、IFN-α+Nimo 50μg/ml组、IFN-α+Nimo 100μg/ml组、IFN-α+Nimo 200μg/ml组72 h时细胞活力分别为1.16±0.09、0.71±0.07、0.88±0.06、0.96±0.08、1.03±0.11；细胞凋亡率分别为（3.51±0.32）%、（21.84±2.23）%、（14.39±1.15）%、（9.18±1.03）%、（8.37±0.94）%;IRS-1蛋白表达水平分别为0.67±0.06、0.19±0.03、0.33±0.04、0.47±0.05、0.59±0.06,NC组、IFN-α组、IFN-α+Nimo 50μg/ml组、IFN-α+Nimo 100μg/ml组、IFN-α+Nimo 200μg/ml组上述指标比较差异有统计学意义（P<0.05）。IFN-α+pcDNA组与IFN-α+pcDNA-IRS-1组72 h时细胞活力分别为0.76±0.06、1.04±0.07；细胞凋亡率分别为（20.75±2.07）%、（11.35±1.43）%,IFN-α+pcDNA组与IFN-α+pcDNA-IRS-1组上述指标比较差异有统计学意义（P<0.05）。IFN-α+Nimo+si-con组与IFN-α+Nimo+si-IRS-1组72 h时细胞活力分别为1.03±0.09、0.79±0.06；细胞凋亡率分别为（10.75±1.08）%、（15.36±1.25）%,IFN-α+Nimo+si-con组与IFN-α+Nimo+si-IRS-1组上述指标比较差异有统计学意义（P<0.05）。结论：尼莫地平可能通过上调IRS-1表达从而影响人脑血管外膜成纤维细胞增殖及凋亡。更多还原

【Abstract】 Objective:To investigate the effects of nimodipine on the cell viability and apoptosis of human cerebral vascular adventitial fibroblasts and its regulatory mechanism on IRS-1.Methods:Human cerebrovascular adventitial fibroblasts were cultured in viro,divided to NC group,IFN-α group and co-treatment group, and co-treatment group treated with nimodipine at different concentrations(50,100,200 μg/ml)and IFN-α,which were recorded as IFN-α+Nimo 50 μg/ml group,IFN-α+Nimo 100 μg/ml group,IFN-α+Nimo 200 μg/ml group. MTT measures was used to detect cell viability. Flow cytometry was used to detect the apoptotic rate. qRT-PCR and Western blot were used to detect the expression of IRS-1. pcDNA and pcDNA-IRS-1 were transfected into cells,respectively.si-con and si-IRS-1 were transfected into cells respectively and then treated cells with nimodipine,the above methods were used to detect cell viability and apoptosis rates.Results:In the NC group,IFN-α group,IFN-α+Nimo 50 μg/ml group,IFN-α+Nimo 100 μg/ml group,and IFN-α+Nimo 200 μg/ml group,the cell viability at 72 h were 1.16±0.09,0.71±0.07,0.88±0.06,0.96±0.08,1.03±0.11;the apoptosis rates were(3.51±0.32)%,(21.84±2.23)%,(14.39±1.15)%,(9.18±1.03)%,(8.37±0.94)%;IRS-1 protein expression levels were 0.67±0.06,0.19±0.03,0.33±0.04,0.47±0.05,0.59±0.06,NC group,IFN-α group,IFN-α+Nimo 50 μg/ml group,IFN-α+Nimo 100 μg/ml group,IFN-α+Nimo 200 μg/ml group had statistically significant differences in the above indicators(P<0.05). In IFN-α+pcDNA group and IFN-α+pcDNA-IRS-1 group,cell viability at 72 h were 0.76±0.06,1.04±0.07,and the apoptosis rates were(20.75±2.07)%,(11.35±1.43)%,there was a statistically significant difference in the above indexes between the IFN-α+pcDNA group and IFN-α+pcDNA-IRS-1 group(P<0.05). The cell viability at 72 h in IFN-α+Nimo+si-con group and the IFN-α+Nimo+si-IRS-1 group were 1.03±0.09 and 0.79±0.06,respectively;the apoptosis rates were(10.75±1.08)%,(15.36±1.25)%,IFN-α+Nimo+si-con group and the IFN-α+Nimo+si-IRS-1 group were statistically different(P<0.05).Conclusion:Nimodipine may affect the proliferation and apoptosis of human cerebral vascular adventitial fibroblasts by up-regulating the expression of IRS-1.更多还原