溶血磷脂酸对THP-1细胞Toll样受体4表达的影响

Effect of lysophosphatidie acid on the expression of toll like receptor 4 in THP-1 cell

目的观察溶血磷脂酸(lyso phosphatidie acid,LPA)对人单核细胞株THP-1细胞Toll-样受体4(toll like receptor 4,TLR4)mRNA、核蛋白NF-κBp65表达的影响以及细胞因子TNF-α的变化,探讨TLR4/NF-κB在LPA致动脉粥样硬化中的作用。方法以不同浓度水平LPA(0-10μM)刺激人THP-1细胞4h,RT-PCR法测定TLR4mRNA表达,Westernblot检测核蛋白NF-κBp65表达变化,ELISA法测定细胞因子TNF-α。结果LPA以剂量依赖的方式上调THP-1细胞TLR4表达,激活NF-κB,促进TNF-α分泌。结论LPA致粥样硬化作用可能部分是由TLR4/NF-κB信号途径介导的,干预TLR4/NF-κB可能抗粥样硬化治疗的一个新的靶点。

[ Objective ] To study the effect of lysophosphatidie acid （LPA） on the expression of toll like receptor 4 （TLR-4） mRNA, NF-κB p65 protein in cell nucleus and the secretion of TNF-α in THP-1 cell line, and to investigate the role of TLR4 / NF-κB invovled in the atheroselerosis caused by LPA. [Methodes] Human THP-leells were stimulated with LPA at different concentrations （0-10 μM）, TLR-4 mRNA was measured by RT-PCR, the activity of NF-κB p65 subunit was detected by Westernblot according to the expression level of NF-κB p65 protein in cell nucleus, the secretion of TNF-α was detected using a sandwich ELISA. [Results] LPA upregulated the expression of TLR-4, activated the NF-κB p65, increased the level of TNF-α in a dose-dependent manner, the peak point of TLR-4 and NF-κB p65 occurred at 1 μM LPA and decreased at higher LPA concentrations. [Conclusions] The atherogenie effect of LPA maybe partially mediated through TLR4/NF-κB pathway, which may provide a target for the treatment of atherosclerosis.