脂氧素A4对TNF-α诱导的内皮细胞炎症反应的影响

Effects of Lipoxin A4 on the inflammatory reaction induced by tumor necrosis factor-α in endothelial cells

目的:探讨脂氧素A4(LXA4)对肿瘤坏死因子-α(TNF-α)诱导的人肺微血管内皮细胞(HPMECs)炎症相关因子的影响,并初步探讨其可能的机制。方法:实验分为对照组、TNF-α单独刺激组和不同浓度LXA4(5 ng/mL、25 ng/mL、50 ng/mL)预处理组。实时荧光定量PCR检测HPMECs单核细胞趋化蛋白-1(MCP-1)、E-选择素(E-selectin)、白介素-6(IL-6)mRNA表达水平;细胞免疫化学方法定位检测核因子-κB/p65(NF-κB/p65);蛋白质免疫印迹(Western blot)方法检测细胞核内和细胞总蛋白中NF-κB/p65的水平。结果:LXA4可以抑制TNF-α诱导的HPMECs MCP-1、E-selectin、IL-6 mRNA表达的上调;LXA4可以抑制TNF-α引起的HPMECs p65由细胞质向细胞核的转位。结论:LXA4可能通过抑制NF-κB细胞信号通路而抑制血管内皮细胞炎症相关因子的表达,发挥抗炎作用。

Objective： To investigate the effects of Lipoxin A4 （LXA4） on inflammatory related factors induced by tumor necrosis factors （TNF-α） in human pulmonary microvascular endothelial cells （HPMECs）. Methods： HPMECs were treated with LXA4 （5, 25 and 50 ng/mL） in various concentrations prior to TNF- α （50 ng/mL） exposure. Monocyte chemoattractant protein-1 （MCP-1）, E-selectin and Interleukin-6 （IL-6） were ana- lyzed with real-time quantitative reverse transcription polymerase chain reaction （real-time RT-PCR）. The effects of LXA4 on TNF- c（ -induced p65 subgroup of nuclear factor-k B （NF-kB） nuclear translocation were observed with immuncytochemistry and western blot. Results： The mRNA expression for MCP-1, E-selectin and IL-6 in response to TNF- c（ were decreased by LXA4. LXA4 significantly reduced nuclear translocation of NF- K B p65 in HPMECs induced by TNF- α. Conclusion： LXA4 may inhibit the expression of inflammatory related factors at least in part via NF- KB signaling pathways to play a role in anti-inflammation.