尼莫地平调控IRS-1对人脑血管外膜成纤维细胞活力和凋亡的影响  

Effects of nimodipine on IRS-1 on cell viability and apoptosis of human cerebral vascular adventitial fibroblasts

作　　者：袁岗[1] 肖宗宇(指导)[1] 李文辉[1] 曹立新[1] 惠超杰 YUAN Gang;XIAO Zongyu;LI Wenhui;CAO Lixin;HUI Chaojie(Department of Neurosurgery,Affiliated Hospital of Qinghai University,Xining 810001,China)

机构地区：[1]青海大学附属医院神经外科,西宁810001

出　　处：《中国免疫学杂志》2022年第10期1171-1176,共6页Chinese Journal of Immunology

基　　金：青海省自然科学基金青年项目(2015-ZJ-943Q);青海大学附属医院中青年科研基金重点项目(ASRF-ZD-01)。

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。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

关 键 词：尼莫地平 IRS-1 人脑血管外膜成纤维细胞 细胞活力 凋亡 

分 类 号：R363[医药卫生—病理学]