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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 the inflammatory cytokines induced by ox-LDL in EA.hy926 cells and to explore its possible mechanism from the activation of nuclear factor-κB (NF-κB). Methods: The normal cultured EA.hy926 cells were randomly divided into blank group, catalpol group (0.5 mmol·L -1), ox-LDL group (100 mg · L -1), catalpol high and low dose group , 0.05 mmol·L-1). The drug groups were incubated for 24 h, and the blank group and the ox-LDL group were incubated with blank medium for 24 h. After incubation, 100 mg · L -1 ox-LDL was incubated for 24 h. The expressions of tumor necrosis factor-α (TNF-α) and vascular cell adhesion molecule-1 (VCAM-1) mRNA and protein were detected by reverse transcription-polymerase chain reaction (RT-PCR) and Western blotting Cell viability was detected by MTT assay. Apoptosis rate was detected by nuclear staining of Hoechst cells. The activation of NF-κB was detected by Western blot and enzyme-linked immunosorbent assay (ELISA). Results: Compared with the blank group, the cell injury of the catalpol protective group was significantly reduced, and the mRNA and protein expressions of TNF-α and VCAM-1 were significantly decreased. The activation of NF-κB was significantly inhibited in a dose-dependent manner (P <0.05). Conclusion Catalpol can effectively reduce the inflammatory response induced by ox-LDL in EA.hy926 cells and protect EA.hy926 cells, which may be related to the inhibition of NF-κB activation.