Study of angiotensin-Ⅱactivating NF-κB in human pulmonary microvascular endothelial cells through classical pathway / 中华急诊医学杂志

ABSTRACT

Objective To investigate the activation of nuclear factor-κB(NF-κB),which was stimulated by angiotensin-Ⅱ(AngⅡ)through classical pathway in human pulmonary microvascular endothehal cells(HPMEC).Method The experiment was divided into two groupsin Ang Ⅱ group,HPMECS were incubated with 10-6mol/L AngⅡ for 0,0.5,1,2 and 4 hours,respectively;in losartan group,HPMEC was pretreated with 10-6mol/L losartan(inhibitor of AngⅡ type 1 receptor)for 1 hour,and then stimulated with 10-6mol/L AngⅡ for 2 hours,and the nucleax protein and the cell plasma protein were prepared by lysis and centrifugation.Electrophoretic mobility shift assay(EMSA)was used to detect the NF-κB DNA binding activity.The inhibitor of κBa(IκBα)was detected by Western blotting.The data were expressed as(x±s)and analyzed with one way analysis of variance.A P value less than 0.05 indicated significant difference.Results Compared with the activity of NF-κB at 0 h (100.0±25.1)after AngⅡstimalation,the activity increased significantly at 0.5 hour(144.5±16.1,P＜0.05),and reached peak value at 2 hours(270.1±27.2,P＜0.05).The concentration of IκBα at 0 hours was 44.4%±2.1%,decreased markedly at 0.5 hours(38.9%±3.6%,P＜0.05),and to the lowest level at 2hours(32.6%±2.3%,P＜0.05).The activity of NF-κB(115.4±10.7)and the concentration of IκBα(43.6%±3.7%)in losartan group had ilo significant difference with AngⅡ group at 0 h(P＞0.05).The activity of NF-κB and the concentration of IκBα in losartan group had significant difference with AngⅡ group at 2hours.Conclusions NF-κB can be activated through classical pathway,which stimulated by AngⅡ in HPMEC.