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目的研究核转录因子NF-κB(Nuclear factor kappa B)在ONO-AE-248诱发中性粒细胞非凋亡、非坏死性死亡中的作用,探讨这种细胞死亡的分子机制。方法Ficoll法分离健康志愿者外周静脉血中性粒细胞,与ONO-AE-248共同培养,通过Western blotting检测IκBα的蛋白表达水平,反映NF-κB的活化状态,并应用RT-PCR检测NF-κB调节的基因XIAP、DAD1和FLIP的mRNA表达水平。结果ONO-AE-248不能促进中性粒细胞内IκBα蛋白降解,显示它并不诱导NF-κB活化。而且,ONO-AE-248对NF-κB调节的基因XIAP、DAD1、FLIP的mRNA表达也无明显影响(P>0.05)。结论ONO-AE-248诱发的中性粒细胞非凋亡、非坏死性死亡不依赖NF-κB信号转导通路,其介导中性粒细胞死亡的信号转导可能是不同于凋亡的独特转导途径。
Objective To investigate the role of nuclear factor kappa B (NF-κB) in non-apoptotic and non-necrotic death induced by ONO-AE-248 in neutrophils and to explore the molecular mechanism of this cell death. Methods Peripheral venous blood neutrophils were isolated from healthy volunteers by Ficoll method and co-cultured with ONO-AE-248. The protein expression of IκBα was detected by Western blotting and the activation state of NF-κB was detected by RT-PCR. κB-regulated gene XIAP, DAD1 and FLIP mRNA expression levels. Results ONO-AE-248 failed to promote neutrophil IκBα protein degradation, indicating that it did not induce NF-κB activation. Moreover, ONO-AE-248 also had no significant effect on mRNA expression of NF-κB-regulated genes XIAP, DAD1 and FLIP (P> 0.05). Conclusions ONO-AE-248-induced neutrophil non-apoptotic and non-necrotic death is not dependent on NF-κB signaling pathway. The signal transduction of neutrophil death may be distinct from that of apoptosis Transduction pathway.