脂氧素A4对脂多糖作用RAW264.7细胞的实验研究

Preliminary study on effect of LXA4 on LPS-induced RAW264.7 Macrophages

目的探讨脂氧素A4(LXA4)对脂多糖(LPS)作用巨噬细胞存活的影响作用。方法取对数生长期的鼠源性RAW264.7巨噬细胞株进行培养、传代。采用免疫印迹法(Western blot)检测不同浓度LPS处理巨噬细胞的pNF-κB p65蛋白水平,并应用CCK-8法观察LPS对巨噬细胞的毒性作用及LXA4对其存活率的影响。结果 LPS对巨噬细胞内pNF-κB p65蛋白水平存在影响,且在一定范围内呈剂量依赖关系;LXA4能够逆转LPS对巨噬细胞的毒性作用。结论 LXA4能够抑制LPS对巨噬细胞的作用及NF-κB的激活,从而减轻炎症反应。

Objective To examine the effect of lipoxin A4 （LXA4） on the survival of macrophage.Methods Macrophages were treated with different concentrations of lipopolysaccharide （LPS） for 2-,4-and 6-h,respectively.The nuclear protein was isolated from the cells and the level of pNF-κB in macrophages was detected by Western Blot.Then the cells were cultured with or without 300 nM LXA4.Cytotoxicity was detected by CCK-8.We also observed the effect of LXA4 on the survival of macrophage,as well as its potential mechanism.Results LPS can promote the translocation of NF-κB in a concentration-dcpcndent way.LXA4 can reverse the cytotoxicity of LPS.Conclusion LXA4 may inhibit the cytotoxicity of LPS and the nuclear translocation of NF-κB,and thereby reduce the inflammation.