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目的:研究星形孢菌素(staurosporine,STS)诱导的小鼠T细胞中Caspase非依赖性细胞死亡方式,探讨T细胞死亡的分子机制。方法:在加入或不加Caspase抑制剂z-VAD-fmk(20μmol.L-1)情况下,检测STS(100 nmo.lL-1)刺激后小鼠T细胞的凋亡率,DNA ladder,线粒体膜电位变化以及凋亡相关蛋白AIF,细胞色素C和Bax等的变化。结果:STS诱导的小鼠T细胞凋亡中存在Caspase非依赖性细胞死亡方式,其DNA ladder改变与Caspase依赖性死亡方式不同,线粒体膜电位依然下降,AIF从线粒体转移到细胞质中要早于细胞色素C,且两者的转移需要Bax结合到线粒体上。结论:在STS诱导的小鼠T细胞死亡中,Caspase非依赖性死亡方式可能是Caspase依赖性死亡方式的替代途径,这对于维持T细胞的数量和保证免疫系统功能的正常发挥具有十分重要的意义。
Objective: To study the caspase-independent cell death in mouse T cells induced by staurosporine (STS) and to explore the molecular mechanism of T cell death. Methods: The apoptosis rate of T lymphocytes was detected by the stimulation of 100 nmo.lL-1 STS with or without Caspase inhibitor z-VAD-fmk (20 μmol.L-1). DNA ladder, mitochondria The change of membrane potential and the changes of apoptosis associated protein AIF, cytochrome C and Bax. Results: There was a caspase-independent cell death pattern in STS-induced mouse T cell apoptosis. The change of DNA ladder was different from that of Caspase-dependent death, and mitochondrial membrane potential was still decreased. AIF migrated from mitochondria to cytoplasm earlier than cells Pigment C, and the transfer of both requires that Bax bind to mitochondria. CONCLUSIONS: In the STS-induced mouse T cell death, Caspase-independent death may be an alternative pathway for Caspase-dependent death, which is of great importance in maintaining the number of T cells and ensuring the normal functioning of the immune system .