二十碳五烯酸对脂多糖处理的人脐静脉内皮细胞核转录因子κB的表达及细胞因子分泌的影响

Effects of eicosapentaenoic acid on expression of nuclear factor κB and release of cytokines in human umbilical vein endothelial cells stimulated by lipopolysaccharide

目的:探讨二十碳五烯酸(EPA)对脂多糖(LPS)处理的人脐静脉内皮细胞(HUVECs)核转录因子κB(NF-κB)表达以及VEGF、IL-1α和IL-6分泌的影响。方法:体外生长良好的传代HUVECs分为对照组、LPS组、0.030 g/L EPA+LPS处理组、0.050 g/L EPA+LPS处理组。LPS组仅加入LPS进行培养,EPA处理组先加入2种浓度的EPA(EPA终浓度分别为0.030 g/L和0.050 g/L)培养1 h,再加入LPS进行培养。LPS刺激6 h、12 h、24h后,收集各组上清液,ELISA检测上清液中VEGF、IL-1α和IL-6的分泌量;LPS刺激24 h后的沉淀细胞,用Western blotting法检测HUVECs NF-κB p65的蛋白表达。结果:与对照组相比,LPS刺激后,HUVECs NF-κB p65表达和VEGF、IL-1α、IL-6分泌显著升高(P<0.05)。EPA抑制LPS处理的HUVECs NF-κB p65的蛋白表达及VEGF、IL-1α和IL-6分泌,除LPS刺激后6 h,EPA处理组IL-6的分泌量与LPS组比较无显著差异(P>0.05)外,其余均差异显著(P<0.05)。结论:LPS使HUVECs NF-κB蛋白表达增加,促进其VEGF及细胞因子表达。EPA抑制LPS处理的HUVECs NF-κB的表达,VEGF和细胞因子的分泌,为EPA应用于各种新生血管性疾病和炎症性疾病的防治提供了理论依据。

AIM ： To investigate the effects of eicosapentaenoic acid（EPA） on the expression of nuclear factor kappa B（ NF - KB） and release of vascular endothelial growth factor（ VEGF）, IL - lot and IL - 6 in cultured human um- bilical vein endothelial cells （ HUVECs ） stimulated by lipopolysaccharide （ LPS ）. METHODS ： HUVECs were obtained from cell strain and cultured in vitro. HUVECs were divided into 4 groups： control group, LPS group, 0. 030 g/L EPA treatment group and 0. 050 g/L EPA treatment group. The cells were cultured with LPS alone in LPS group and incubated with EPA for 1 h in the EPA pretreatment groups at the concentrations of 0. 030 g/L and 0. 050 g/L before LPS stimulation. Twenty - four hours after stimulated by LPS, the protein expression of NF - κB p65 in HUVECs were assessed by Western blotting analysis at different time points. The production of VEGF, IL - 1 α and IL - 6 in cultured HUVECs was evaluated by ELISA. The effects of EPA on the protein expression of NF - κB p65 and the production of VEGF, IL - 1 α and IL - 6 in HUVECs challenged by LPS were also determined. RESULTS： Compared with control group, the protein expression of NF- κB p65 was significantly increased in HUVECs induced by LPS and was inhibited by EPA. Compared with control group, the protein expression of VEGF, IL - 1 α and IL - 6 was dramatically increased in HUVECs induced by LPS and most of the increase was inhibited by EPA. CONCLUSION： LPS enhances the protein expression of NF - κB and the release of VEGF, IL - 1 α and IL - 6. EPA inhibits the protein expression of NF - κB, and the production of VEGF and the inflammatory cytokines in cultured HUVECs stimulated by LPS, indicating that EPA may be useful for preventing and trea- ting neovascular and inflammatory diseases.