Shp-2/NF-κB途径介导脂氧素A4对IL-1β促肾小球系膜细胞IL-6合成作用的抑制

目的 :研究脂氧素A4 (LXA4 )是否拮抗IL 1β促进肾小球系膜细胞IL 6合成的作用 ,并探讨LXA4 作用的信号转导机制。方法 :用不同浓度的LXA4 预处理培养的大鼠肾小球系膜细胞 ,再加入IL 1β共同孵育一定时间 ;或单用IL 1β刺激肾小球系膜细胞。用ELISA法检测培养上清中的IL 6蛋白表达量 ;用RT PCR法检测IL 6的mRNA表达量 ;用免疫沉淀与印迹法检测含Src同源区 2 (SH2 )蛋白酪氨酸磷酸酶 - 2 (Shp 2 )的表达 ;用凝胶电泳迁移率试验 (EMSA)检测NF κB的DNA结合活性。结果 :LXA4 呈剂量依赖性地抑制IL 1β诱导的系膜细胞分泌的IL 6蛋白及mRNA表达 ,拮抗IL 1β诱导的Shp 2磷酸化与NF κB的活化。 结论 :LXA4 能够拮抗IL 1β对肾小球系膜细胞IL 6合成的促进作用 ,其机制依赖于Shp 2 NF κB信号转导途径

Shp-2/NF-κB Pathway Mediates the Inhibition of Lipoxin A4 on IL-1β-induced Synthesis of IL-6 in Glomerular Mesangial Cells

Objective: To examine whether lipoxin A4 (LXA4) has an antagonistic effect on IL-1ββ-induced synthesis of IL-6 in glomerular mesangial cells, and to explore the molecular mechanisms of signal pathway in LXA4 actions. Methods: The glomerular mesangial cells of rat were cultured and treated with IL-1β, with or without preincubation with LXA4 at different concentrations. The amount of IL-6 in the supernatant of cells was analyzed by enzymelinked immunosorbent assay(ELISA). The expressions of mRNA of IL-6 were determined by RT-PCR. The expressions of Src homology 2( SH2 ) containing protein-tyrosine phosphatase 2(Shp-2) were assessed by immunoprecipitation and immunoblotting. Activities of DNA-binding of nuclear factor-kappa B(NF-κB) were measured by electrophoretic mobility shift assay(EMSA). Results: IL-1β-stimulated secretion of protein and expression of mRNA of IL-6 in mesangial cells were inhibited by LXA4 in a dose- dependent manner. LXA4 antagonizes the phosphorylation of Shp-2 and activities of NF-κB induced by IL-1β. Conclusion: LXA4 amtagonists IL-1β-induced synthesis of IL-6 in glomerular mesangial cells through the mechanism of Shp-2/NF-κB pathway- dependent signal transduction.