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为探讨接受ONO-AE-248刺激后重要的表达蛋白-A1在中性粒细胞(polymorphonuelear neutrophils,PMN)非凋亡程序化细胞死亡中的的表达及意义,以进一步确定这种新型细胞死亡模式提供线索。采用Ficoll法新鲜分离健康志愿者外周静脉血PMN,与ONO-AE-248共同培养2 h后提取总RNA,并行反转录-PCR检测A1 mRNA的表达;Western blot检测A1蛋白的表达;透射电镜观察线粒体形态结果的变化。结果发现,ONO-AE-248刺激2 h时PMN A1 mRNA的表达明显上调(P<0.01);蛋白印迹法显示A1蛋白的表达增高;透射电镜观察ONO-AE-248刺激下的早期PMN线粒体超微形态结构发生了剧烈改变。提示ONO-AE-248引起A1表达的上调可能是ONO-AE-248诱使中性粒细胞发生非凋亡性程序化死亡的根本原因之一。
To explore the expression and significance of the key protein expressed in ONO-AE-248 after non-apoptotic programmed cell death in polymorphonuelear neutrophils (PMN) to further confirm the novel pattern of cell death Provide clues. Peripheral venous blood PMN was isolated from healthy volunteers by Ficoll method and total RNA was extracted after co-cultured with ONO-AE-248 for 2 h. The expression of A1 mRNA was detected by reverse transcription-polymerase chain reaction (RT-PCR) Observed changes in mitochondrial morphology results. The results showed that the expression of PMN A1 mRNA was significantly up-regulated after ONO-AE-248 stimulation for 2 h (P <0.01). The expression of Al protein was increased by Western blotting. The mitochondrial ultrastructure of PMN induced by ONO-AE-248 Micro-morphological changes have taken place. It is suggested that the up-regulation of A1 expression induced by ONO-AE-248 may be one of the fundamental reasons for non-apoptotic programmed death induced by ONO-AE-248 in neutrophils.