【摘要】 目的:探讨接受ONO-AE-248刺激后重要的差异表达蛋白——凋亡相关斑点样蛋白(Apoptosis-associated speck-like protein containing CARD,ASC)的表达及意义。方法:Ficoll法新鲜分离健康志愿者外周静脉血中性粒细胞(Polymorpho-nuclear neu-trophils,PMN),与ONO-AE-248共同培养,2小时提取总RNA,并行反转录-PCR检测ASC mRNA的表达;12小时提取总蛋白,双向电泳分离后,采用PDQest2-DE软件分析找出差异表达的蛋白质斑点,并用基质辅助激光解吸电离飞行时间串联质谱(MALDI-TOF-MS)进行鉴定。结果:ONO-AE-248刺激2小时中性粒细胞ASC mRNA的表达明显下调(P<0.01);12小时通过双向电泳和蛋白质质谱分析发现ASC为重要的差异表达蛋白且ONO-AE-248刺激后的表达量明显低于自发性凋亡组。结论:ONO-AE-248引起的ASC表达的下调可能是中性粒细胞凋亡和ONO-AE-248诱导的中性粒细胞非凋亡性程序化细胞死亡的关键性差异点。ONO-AE-248显著下调ASC的表达,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。更多还原

【Abstract】 Objective:To investigate the roles of apoptosis-associated speck-like protein containing CARD(ASC),one of differentially expressed proteins in neutrophil death induced by ONO-AE-248.Methods:Human neutrophils were cultured in vitro with or without ONO-AE-248(5×10-5 mol/L).The expression of ASC mRNA was detected by RT-PCR.PMN proteins were extracted at 12 h and then separated by 2D electrophoresis.20 differentially expressed proteins were selected and determined by PDQest 2-DE software.Results:ONO-AE-248 decreased the expression of ASC mRNA strikingly in 2 hours;ASC was one of the important differentially expressed proteins between spontaneous apoptosis and non-apoptosis programmed cell death induced by ONO-AE-248 in 12 hour and ASC protein point was weaker in ONO-AE-248 group.Conclusion:ASC might act as double switch that leads to apoptosis of neutrophils in the high expression and removes the forbiddance of pathway of ONO-AE-248 signals in the low expression,resulting in non-apoptosis programmed cell death of neutrophils.更多还原