NF-κB/IκB在二苯乙烯苷抑制过氧化氢诱导内皮细胞凋亡中的表达变化

摘要

目的 探讨二苯乙烯苷(TSG)对过氧化氢(H2O2)诱导人脐静脉内皮细胞凋亡及对核因子κB(NF-κB)、NF-κB抑制因子(IκB)基因表达的影响.方法 体外培养人脐静脉内皮细胞,实验分为对照组、H2O2组、TSG组,采用Hoechst 33258染色观察细胞凋亡形态,MTT法检测细胞增殖率,流式细胞术检测细胞凋亡率,RT-PCR检测NF-κB与IκB mRNA表达,Western blot检测NF-κB p65、IκB蛋白表达.结果 与对照组比较,H2O2组凋亡细胞数增多,细胞凋亡率增加,细胞增殖降低;NF-κB mRNA和蛋白表达增加,IκB mRNA和蛋白表达降低,差异均有显著性(P<0.01).经TSG预处理后,细胞的增殖率较H2O2组增加,细胞凋亡率减少;NF-κB mRNA和蛋白表达减少,IκB mRNA和蛋白表达增加(P<0.01).结论 二苯乙烯苷能抑制H2O2诱导的人脐静脉内皮细胞凋亡,其作用机制可能与调节NF-κB/ IκB表达有关.%Aim To investigate the effect of 2,3 .5 ,4'tetrahydroxystilbene-2-0- β-D-glucoside ( TSG ) on human umbilical vein endothelial cells ( HUVECs ) apoptosis induced by hydrogen peroxide( H2O2 ) and the expressions of nuclear factor kappaB ( NF-κB ) , inhibitor of NF-κB ( IκB ). Methods HUVECs were divided into three groups: control group, H202 group and TSG group. The morphology of apoptosis was detected via Hoechst33258 stain. The proliferation and apoptosis rates of HUVECs were detected by MTT and Flow Cytometry respectively. The gene expressions of NF-κB,IκB were assessed by RT-PCR and Western blot respectively. Results The proliferation rate of HUVECs decreased significantly compared with control group,while the apoptosis rate increased obviously after treated with H2O2. The proliferation rate of TSG pretreatment group increased significantly while the apoptosis rate decreased obviously compared with H2O2 group.Compared with H2O2 group, the expressions of NF-κB mRNA and protein declined, and the levels of IκB mRNA and protein increased significantly after pretreated with TSG ( P ＜ 0. 01 ). Conclusion TSG can inhibit the apoptosis of HUVECs induced by H2O2, and the molecular mechanism may be associated with regulating the expression of NF-κB/IκB.