论文部分内容阅读
背景他汀类药物有独立于调脂作用之外的抗炎作用。近年研究表明Toll-样受体4(TLR4)参与了动脉粥样硬化的形成和发展。目的观察阿托伐他汀对脂多糖(LPS)诱导的内皮细胞TLR4及其下游分子表达的影响,以探讨他汀类药物抗炎作用的分子机制。方法采用阿托伐他汀(1及10μmol/L)或核转录因子(NF)κB抑制剂咖啡酸苯乙酯(CAPE)预孵育人脐静脉内皮细胞(HUVEC)30min后,应用LPS(1mg/L)作用24h。逆转录聚合酶链反应(RT-PCR)方法检测TLR4、细胞间黏附分子1(ICAM-1)和E选择素mRNA表达水平;采用流式细胞术检测TLR4蛋白表达水平;采用蛋白质印迹技术检测核蛋白NF-κBp65表达的变化。结果与LPS组比较,阿托伐他汀1μmol/L组减轻LPS介导的TLR4表达增加[TLR4mRNA:(1.24±0.21)比LPS组(1.82±0.27),P<0.05;TLR4阳性细胞数(50.1±4.7)%比LPS组(69.5±7.8)%,P<0.05],阿托伐他汀减轻LPS介导的ICAM-1和E选择素的表达增加。阿托伐他汀抑制LPS介导的NF-κBp65活化(50.4±10.1比LPS组72.3±12.5,P<0.05),10μmol/L阿托伐他汀较1μmol/L作用更明显;CAPE(20mg/L)也明显抑制了LPS介导的TLR4及ICAM-1和E选择素表达上调。结论阿托伐他汀抑制TLR4/NF-κB及其下游分子表达是他汀类药物抗炎作用机制之一。
Background Statins have an anti-inflammatory effect independent of lipid-lowering effects. In recent years, studies have shown that Toll-like receptor 4 (TLR4) is involved in the formation and development of atherosclerosis. Objective To observe the effect of atorvastatin on the expression of TLR4 and its downstream molecules induced by lipopolysaccharide (LPS) in endothelial cells to explore the molecular mechanism of anti-inflammatory effects of statins. Methods Human umbilical vein endothelial cells (HUVECs) were pre-incubated with atorvastatin (1 and 10 μmol / L) or NF-κB inhibitor caffeic acid phenethyl ester (CAPE) for 30 min. ) Effect 24h. The expression of TLR4, ICAM-1 and E-selectin mRNA were detected by reverse transcription polymerase chain reaction (RT-PCR). The expression of TLR4 protein was detected by flow cytometry. The protein expression of TLR4 was detected by Western blotting Changes of protein NF-κBp65 expression. Results Compared with LPS group, 1μmol / L of atorvastatin could reduce the LPS-induced increase of TLR4 expression (1.24 ± 0.21 vs 1.82 ± 0.27, P <0.05), the number of TLR4 positive cells (50.1 ± 4.7%) compared with LPS group (69.5 ± 7.8)%, P <0.05]. Atorvastatin attenuated LPS-induced increase of ICAM-1 and E-selectin expression. Atorvastatin inhibited LPS-mediated activation of NF-κBp65 (50.4 ± 10.1 vs 72.3 ± 12.5, P <0.05), and atorvastatin at 10 μmol / L had a more pronounced effect than at 1 μmol / L. CAPE (20 mg / Also significantly inhibited LPS-mediated TLR4 and ICAM-1 and E-selectin expression upregulation. Conclusions Atorvastatin inhibits the expression of TLR4 / NF-κB and its downstream molecules is one of the anti-inflammatory mechanisms of statins.