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目的 :研究脂氧素A4 (LXA4 )是否抑制肿瘤坏死因子 α(TNF α)所致的肾小球系膜细胞增殖 ,并探讨LXA4 作用的信号转导机制。方法 :用不同浓度的脂氧素A4预外理培养的大鼠肾小球系膜细胞 ,再加入TNF α(10ng ml)共同孵育 ;或单用TNF α刺激肾小球系膜细胞。用四甲基偶氮唑蓝 (MTT)法检测细胞增殖 ;用RT PCR法检测细胞周期素E与mRNA表达 ;用Westernblot检测 30 8位苏氨酸(Thr30 8)磷酸化的蛋白激酶Akt1及细胞周期负调控蛋白P2 7kipl表达量。结果 :LXA4 呈剂量依赖性地抑制TNF α诱导的系膜细胞增殖。在系膜细胞增殖同时TNF α引起的细胞周期素E的蛋白表达也被LXA4 下调。LXA4 减少TNF α诱导的肾小球系膜细胞中 30 8位苏氨酸磷酸化的Akt1蛋白。LXA4 呈剂量依赖性地阻止TNF α所致的P2 7kipl表达下调。结论 :LXA4 能够抑制TNF α所致的大鼠系膜细胞的增殖 ,其机制依赖于Akt1 P2 7kipl信号转导途径。
AIM: To investigate whether lipoxin A4 (LXA4) inhibits the proliferation of glomerular mesangial cells induced by tumor necrosis factor α (TNFα) and explores the signal transduction mechanism of LXA4. Methods: Rat glomerular mesangial cells were pre-cultured with different concentrations of lipoxin A4 and incubated with TNFα (10ng ml), or mesangial cells were stimulated with TNFα alone. The cell proliferation was detected by MTT assay. The expression of cyclin E and mRNA was detected by RT-PCR. The protein kinase Akt1 and Thr308 phosphorylated protein kinase Akt1 Cycle negative regulatory protein P2 7kipl expression levels. Results: LXA4 inhibited TNFα-induced mesangial cell proliferation in a dose-dependent manner. TNF-alpha-induced cyclin E protein expression was also down-regulated by LXA4 in mesangial cells. LXA4 decreases TNFα-induced 30th threonine phosphorylation of Akt1 protein in mesangial cells. LXA4 dose-dependently prevented down-regulation of P2? Kipl expression by TNF ?. Conclusion: LXA4 can inhibit the proliferation of rat mesangial cells induced by TNFα, the mechanism of which depends on the Akt1 P2 7kip1 signal transduction pathway.