骨膜蛋白siRNA转染抑制ox-LDL诱导的人主动脉内皮细胞系损伤

目的探讨骨膜蛋白(Postn)对氧化型低密度脂蛋白(ox-LDL)诱导的人主动脉内皮细胞系(HAECs)损伤的影响及其作用机制。方法将HAECs随机分为4组:对照组、ox-LDL组、Postn siRNA组和negative siRNA组。RT-q PCR检测mRNA水平;Western blot检测蛋白表达;MTT检测细胞增殖;流式细胞仪检测细胞凋亡;EMSA检测NF-κB的DNA结合能力。结果与对照组相比,ox-LDL组Postn的mRNA和蛋白水平显著提高(P0.05);细胞增殖能力降低(P0.05);细胞凋亡率升高(P0.05);VCAM1、ICAM1、E-selectin、IL-1β、IL-6、TNF-α、p65和p-IκB-α的蛋白表达水平显著上调(P0.05),且NF-κB的DNA结合能力提高(P0.05),Postn siRNA转染能够逆转以上结果。结论 Postn siRNA转染能够抑制ox-LDL诱导的内皮细胞损伤,这可能与抑制NF-κB信号通路有关。

Periostin siRNA transfection inhibits ox-LDL-induced injury in human aortic endothelial cell line

Objective To investigate the effects of periostin (Postn) on oxidized low density lipoprotein (ox-LDL)-induced injury in human artery endothelial cells (HAECs) and its underlying mechanisms.Methods The HAECs were randomly divided into 4 groups:control group,ox-LDL group,Postn siRNA group and negative siRNA group.The mRNA and protein expression was analyzed by RT-qPCR and Westem blot respectively.Cell proliferation was tested by MTT.Cell apoptosis was determined by flow cytometry.NF-κB DNA binding ability was measured by EMSA.Results Compared with control group,the mRNA and protein levels of Postn were increased significantly (P＜0.05);the ability of cell proliferation was reduced (P＜0.05);the cell apoptosis rate was increased (P＜ 0.05);the protein expression levels of VCAM1,ICAM1,E-selectin,IL-1β,IL-6,TNF-α,p65 and p-IκB-α were significantly up-regulated (P＜0.05),and the NF-κB DNA binding ability was markedly increased (P＜ 0.05) in ox-LDL group,which were all reversed in Postn siRNA group.Conclusions Postn siRNA transfection may reduce ox-LDL-induced endothelial cell injury,which may be related with the inhibition of NF-κB signaling pathway.