氟伐他汀抑制抗β_2GPI/β_2GPI复合物诱导的THP-1细胞活化

Inhibitory effects of fluvastatin on activation of THP-1 cells induced by anti-β_2GPI/β_2GPI complex

为了探讨氟伐他汀对抗β2GPI/β2GPI复合物诱导THP-1细胞活化的干预作用及机制,采用脂多糖(LPS,500μg·L-1)、抗β2GPI/β2GPI复合物(100 mg·L-1)等活化单核细胞株THP-1并用氟伐他汀(0、1、5、10及50 mg·L-1)对细胞进行预处理。采用实时定量PCR(RT-qPCR)检测细胞组织因子(TF)及肿瘤坏死因子-α(TNF-α)mRNA表达,发色底物法检测TF活性,ELISA方法测定TNF-α蛋白的表达,蛋白印迹检测细胞NF-κB抑制蛋白(IκB-α)和磷酸化NF-κB p65的蛋白水平变化。结果显示,抗β2GPI/β2GPI复合物(100 mg·L-1)能诱导THP-1细胞活化以及TF和TNF-α表达上调。氟伐他汀(50 mg·L-1)能抑制复合物诱导的THP-1细胞TF(mRNA和活性)及TNF-α(mRNA和蛋白)的表达(P<0.05),上调IκB-α表达及抑制NF-κB p65磷酸化。结果表明,氟伐他汀通过干预信号分子IκB-α和磷酸化NF-κB p65的表达,抑制抗β2GPI/β2GPI复合物诱导的细胞TF及TNF-α的表达,抑制THP-1细胞活化。

This study is to explore the interventional effects of fluvastatin on anti-β2GPI/β2GPI-induced activation in THP-1 mononuclear cells. In vitro, human mononuclear cells THP-1 were treated with fluvastatin, LPS and anti-β2GPI/β2GPI, then the TF expression on THP-1 cells was detected by real-time quantitative PCR （RT-qPCR） or TF activity was detected by kit. TNF-α mRNA and its protein expression were investigated by RT-PCR and ELISA kit. The expression of phospho-NF-κB p65 and inhibitory protein of NF-κB （IκB-α） were measured by Western blotting. The results suggested that the expression of TF and TNF-α on THP-1 cells was significantly up-regulated with treatment of anti-β2GPI/β2GPI complex （100 mg·L-1）, compared with that of untreated cells （P 〈 0.05）. Fluvastatin （50 mg·L-1） could decrease TF （mRNA and activity） expression and the level of TNF-α （mRNA and protein） in THP-1 cells with anti-β2GPI/β2GPI complex. The expression of TF and TNF-α was shown in a concentration-dependent manner. Moreover, anti-β2GPI/β2GPI complex could down-regulate IκB-α levels and increase the levels of phospho-NF-κB p65. And these effects of anti-β2GPI/β2GPI complex could be blocked by fluvastatin. In conclusion, fluvastatin may interfere the expression and regulation of NF-κB signal transduction pathway, thereby inhibit the effects of anti-β2GPI/β2GPI on activation of THP-1 cells, by decreasing the expression of TF and TNF-α.