目的:研究二十碳五烯酸(eicosapentaenoic acid,EPA)对THP.1巨噬细胞源性泡沫细胞形成的影响。方法:将THP-1单核细胞、佛波酯(phorbolmyristate acetate,PMA)和氧化型低密度脂蛋白(oxidized low density lipoprotein,ox-LDL)共同培养建立泡沫细胞模型,以不同浓度(200、100、50μmol/L)的EPA处理细胞48h后,采用酶法测定细胞内总胆固醇(total cholesterol,TC)及胆固醇酯(cholesterol ester,CE)含量,采用酶联免疫吸附实验(euzyme-linked immunosorbent assay,ELISA)测定细胞上清液中白细胞介素-6(interleukin-6,IL-6)及肿瘤坏死因子-α(tumornecrosisfactor-α,TNF-α)的含量,采用半定量反转录聚合酶链式反应(reverse transcript polymerase chain reaction,RT-PCR)测定三磷酸腺苷结合盒转运体A1(ATP binding cassette transporter A1,ABCA1)mRNA表达水平。结果:与模型组比较,EPA干预组细胞内TC及CE含量均降低,并存在显著差异(P〈0.05);细胞上清液中IL-6及TNF-α含量也显著降低(P〈0.05),并具有量一效关系;低浓度的EPA具有上调ABCA1 mRNA表达的作用。结论:EPA能够降低泡沫细胞内TC的蓄积,抑制泡沫细胞的形成,降低IL-6及TNF-α炎性因子的分泌水平,能够上调ABCA1 mRNA表达水平,进而预防动脉粥样硬化的发生发展。
Objective: To investigate the effect of eicosapetaenoic acid (EPA) on the formation of THP-1 monocyte-derived foam cells. Methods: THP-1 monocytes were cultured with phorbol myristate acetate (PMA) and oxidized low-density lipoprotein (ox-LDL), and intervened by EPA at concentrations of 200, 100 and 50 μmol/L for 48 hours. The cholesterol content of foam ceils was measured by cholesterol kit. The concentrations of IL-6 and TNF-a in culture supernatant were detected by ELISA. The expression of ATP-binding cassette transporter A1 (ABCA1) was detected by semi-quantitative reverse transcription-polymerase chain reaction (RT-PCR). Results: EPA decreased the intracellular concentration of cholesterol (P 〈 0.05) when compared with the control group. EPA decreased the concentrations of IL-6 and TNF-α in culture supernatant (P 〈 0.05) in a dose-dependent manner and low-dose EPA increased the expression of ABCA1 mRNA. Conclusion: EPA reduces the accumulation of intracellular cholesterol and inhibits the transformation of macrophages into foam cells. EPA decreases the secretion of inflammatory factor and low-dose of EPA increases the expression of ABCA1 mRNA, which may partially explain its anti-atherosclerotic activity.