梓醇对NF-κB活化减轻ox-LDL诱导的内皮细胞损伤的影响

Effect of Catalpol on NF-κB Activation and Reducing Endothelial Cells Injury Induced by ox-LDL

目的:观察梓醇对氧化低密度脂蛋白(ox-LDL)诱导EA.hy926细胞炎症因子的影响,并从核因子-κB(NF-κB)的活化角度探讨其可能的作用机制。方法:将常规培养的EA.hy926细胞随机分为空白组,梓醇组(0.5 mmol·L-1),ox-LDL组(100 mg·L-1),梓醇高、低剂量组(0.5,0.05 mmol·L-1),各药物组先孵育24 h,空白组与ox-LDL组给予空白配养基孵育24 h,孵育后加入100 mg·L-1ox-LDL继续培养24 h。逆转录-聚合酶链反应(RT-PCR)和蛋白免疫印迹(Western blot)检测细胞中肿瘤坏死因子-α(TNF-α),血管细胞黏附分子-1(VCAM-1)mRNA和蛋白的表达;噻唑蓝(MTT)法检测细胞活性;Hoechst细胞核染色检测细胞凋亡比率;Western blot和酶联免疫吸附测定(ELISA)检测NF-κB的活化。结果:与空白组比较,梓醇保护组细胞损伤明显减轻,TNF-α,VCAM-1 mRNA及蛋白表达均显著降低,NF-κB活化明显受抑制,且呈剂量依赖性(P<0.05)。结论:梓醇能够有效减轻ox-LDL诱导EA.hy926细胞炎症反应,保护EA.hy926细胞,其机制可能与其抑制NF-κB活化有关。

Objective： To observe the effect of catalpol on oxidized low density lipoprotein（ ox-LDL）induced inflammatory cytokine in EA. hy926 cells and to explore its possible mechanism from the prospective of nuclear factor-κB（ NF-κB） activation. Method： Humane endothelial cell line EA. hy926 was cultured and randomly divided into blank group,catalpol group（ 0. 5 mmol·L- 1）,ox-LDL group（ 100 mg·L- 1）,catalpol high-dose group（ 0. 5 mmol·L- 1）,and low-dose group（ 0. 05 mmol·L- 1）. The mRNA and protein expression levels of tumor necrosis factor α（ TNF-α） and vascuolar cell adhesion molecule-1（ VCAM-1） in EA. hy926 cells were detected by RT-PCR and Western blot respectively. Viability and apoptosis ratio of EA. hy926 cells were detected by MTT and Hoechst staining respectively. NF-κB activation was detected by Western blot and ELISA.Result： As compared with the blank group,EA. hy926 cells were injured very slightly in catalpol protection group,and the mRNA and protein expression levels of TNF-α and VCAM-1 were significantly reduced and NF-κB activation was inhibited in a dose-dependent manner（ P 0. 05）. Therefore,catalpol can effectively reduce ox-LDL-induced inflammatory response in EA. hy926 cells, which may be related to inhibition of NF-κB activation.