目的研究线粒体在ONO—AE-248诱导中性粒细胞非凋亡性程序化细胞死亡中形态结构及功能的变化情况,为进一步确立这一新的死亡形式和找寻其独特的死亡信号转导途径提供实验依据。方法体外新鲜分离人外周血中性粒细胞与ONO—AE-248共同培养,流式细胞术检测其线粒体膜势能的变化;激光共聚焦扫描显微镜观察线粒体形态结构和分布情况。结果ONO—AE-248迅速导致中性粒细胞线粒体膜势能的溃散;ONO—AE-248刺激后中性粒细胞在线粒体形态、结构、分布和胞浆内染色特性等方面均与阴性对照有明显差异。结论线粒体变化是ONO—AE-248诱导中性粒细胞非凋亡性程序化细胞死亡早期形态学上区别于凋亡的显著变化之一。线粒体膜通透性转变以及线粒体膜势能下降可能是ONO—AE-248诱导的非凋亡性程序化细胞死亡的主要原因。
Objective To investigate the changes and roles of mitochondria in neutrophil death induced by ONO-AE-248. Methods Human neutrophils were cultured in vitro with ONO-AE-248(5 - 10,5 mol/L)and medium, respectively. FACS and confocal microscopy were used to detect the mitochondria structure and flow cytometry was employed to detect the level of mitochondria membrane potential. Resuits ONO-AE-248 resulted in a rapid loss of mitochondrial membrane potential of neutrophils. The mitochondrial structure and distribution also were obviously different between spontaneous apoptosis and non-apoptosis programmed cell death induced by ONO-AE-248. Conclusion The experiment results suggest that changes of mitochondrial structure and function might be a typical morphological, physiological, and biochemical feature in this unique form of neutrophil death, and that the mitochondrial pathway might play an important role in ONO-AE-248-induced death of neutrophils.