脂氧素 A4对尿酸诱导的人脐静脉内皮细胞炎症反应的影响

Effects of lipoxin A4 on the inflammatory reaction induced by uric acid in human umbilical vein endothelial cells

目的：探讨脂氧素A4对尿酸诱导的人脐静脉内皮细胞（ HUVECs）炎症相关因子的影响及其可能的机制。方法实验分为对照组、尿酸单独刺激组和不同浓度脂氧素A4（5、25、50 ng/ml）预处理组。ELISA法检测细胞培养上清中白细胞介素（ IL）-1β、IL-6和肿瘤坏死因子α（ TNF-α）的浓度；实时荧光定量PCR检测HUVECs TNF-α、IL-1β、IL-6 mRNA 表达水平；Western 印迹法检测 p38丝裂原活化蛋白激酶（ MAPK）和NF-κB/p65磷酸化水平。结果12 mg/dl的尿酸刺激HUVECs 24 h,细胞培养上清中的TNF-α、IL-1β和IL-6水平均显著升高（P〈0.01）,脂氧素A4呈浓度依赖性地抑制尿酸诱导的TNF-α、IL-1β和IL-6产生（P〈0.01）。尿酸刺激的TNF-α、IL-1β和IL-6 mRNA表达也被5、25、50 ng/ml脂氧素A4显著下调（P〈0.05）。 Western印迹法证实12 mg/dl的尿酸可诱导HUVECs p38 MAPK和NF-κB/p65蛋白磷酸化水平增加（P〈0.05）,而50 ng/ml脂氧素A4预处理后能够显著降低HUVECs的p38 MAPK和NF-κB/p65磷酸化水平（P〈0.01）。结论脂氧素A4可通过降低p38 MAPK和NF-κB/p65蛋白磷酸化水平而抑制尿酸诱导的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 uric acid（ UA） in human umbilical vein endothelial cells（ HUVECs） . Methods HUVECs were treated with 5, 25, and 50 ng/ml LXA4 prior to exposure to 12 mg/dl UA. Tumor necrosis factor-alpha（TNF-α）, interleukin （ IL）-1β, and IL-6 were analyzed with ELISA and realtime PCR. The phosphorylation levels of p38 mitogen-activated protein kinases（ MAPK） and NF-κB/p65 were observed with Western blot. Results Stimulation of HUVECs with 12 mg/dl UA markedly increased TNF-α, IL-1β, and IL-6 production（P〈0. 01）. Pretreatment with LXA4 significantly inhibited UA-induced production of TNF-α, IL-1β, and IL-6 in a concentration dependent manner（P〈0. 01）. The mRNA expressions of TNF-α, IL-1β, and IL-6 in response to UA were also decreased by LXA4（P〈0. 05）. Western blot analysis showed that the phosphorylation levels of p38 MAPK and NF-κB/p65 were significantly raised by 12 mg/dl UA in HUVECs（P〈0. 05）, but attenuated significantly in the presence of 50 ng/ml LXA4. Conclusion LXA4 may inhibit the expressions of inflammatory related factors induced by UA via reducing p38 MAPK and NF-κB/p65 phosphorylation and play a role in anti-inflammation.