二十碳五烯酸对THP-1巨噬细胞源性泡沫细胞形成的影响及机制

Effect of Eicosapetaenoic Acid on THP-1 Macrophage-Derived Foam Cell Formation and Its Mechanism of Action

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(phorbol myristate 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48 h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(tumor necrosis factor-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)m RNA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-效关系;低浓度的EPA具有上调ABCA1 m RNA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 m RNA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.