ONO-AE-248诱导的中性粒细胞非凋亡性程序化细胞死亡的形态学变化  

Morphological changes of non-apoptotic programmed cell death of polymorphonuclar neutrophils induced by ONO-AE-248

作　　者：张姿[1] 刘佳佳[1] 姚富丽[1] 何浩[1] 羊建[1] 谢华福[1] 何涛[1] 

机构地区：[1]泸州医学院分子生物学实验室,四川泸州646000

出　　处：《细胞与分子免疫学杂志》2007年第5期413-415,共3页Chinese Journal of Cellular and Molecular Immunology

基　　金：国家自然科学基金资助项目(30371305)

摘　　要：目的研究前列腺素E受体亚型3(EP3)激动剂ONO-AE-248诱导的中性粒细胞(PMN)非凋亡性程序化细胞死亡的形态学变化,探讨PMN死亡形式的多样性和复杂性。方法运用透射电镜、荧光显微镜及激光共聚焦扫描显微镜观察PMN在自发性凋亡与ONO-AE-248刺激下亚细胞微结构的变化。结果以5mmol/L的ONO-AE-248刺激PMN12h后,电镜观察绝大多数PMN的多叶核融合为单叶核,核质疏松,核膜分层、起泡甚至破裂,核质溢出,但细胞膜较完整,细胞器无明显改变。以荧光染料DAPI标染活细胞核,ONO-AE-248刺激后,PMN表现为多叶核融合成大而模糊的球形,核染色密度较低;ONO-AE-248刺激的PMN死亡仍有细胞膜磷脂酰丝氨酸(PS)的外翻。ONO-AE-248刺激组与自发性凋亡组PMN在线粒体的形态结构、分布和细胞质内的染色性均有明显差异。结论ONO-AE-248在体外能刺激人PMN发生非凋亡性程序化细胞死亡,这一刺激过程不影响自发性凋亡的PMN细胞膜PS的外翻,并且细胞核和线粒体的改变是其早期最为显著的形态学改变,提示二者可能是ONO-AE-248刺激的靶标和早期的药物反应中心。AIM： To investigate the morphological changes of polymorphonuclar neutrophils （PMNs） induced by ONO-AE-248, a selective agonist of EP3. METHODS： The morphological changes of PMNs treated with or without ONO-AE-248 were observed by electron microscope, fluorescence microscope and confocal microscope. RESULTS： A unique form of PMNs death was rapidly caused by ONO-AE-248. The agonist primarily induced morphological changes of PMN nucleus under electron microscope, including the fusion of the nuclear Iobules same as a large round structure, decreasing of the compactness of chromatin, the blebbing and rupture of nuclear membrance. Observation of PMN nucleus under fluorescence microscope by DAPI staining gave the same conclusion. However, there were no apparent changes in other intracellular organelles. The structure and distribution of mitochondria of PMNs treated with ONO-AE-248 were different from the typical morphological changes of apoptotic PMNs. ONO-AE-248 exerted little effects on the exposure of cell membrane phosphatidylserine （PS）. CONCLUSION： ONO-AE-248 can promote the non-apoptotic programmed cell death of PMNs in vitro. The events occurring in nucleus and mitochondria might be the early features of the novel cell death, which suggests that nucleus and mitochondria may be the reaction center of PMNs induced by ONO-AE-248.

关 键 词：ONO-AE-248 中性粒细胞 线粒体 凋亡 非凋亡性程序化细胞死亡 

分 类 号：R329.12[医药卫生—人体解剖和组织胚胎学]