【摘要】 目的研究鲎素诱导HL-60细胞凋亡时,半胱氨酸蛋白酶(Caspases)活性的变化,探讨鲎素诱导HL-60细胞凋亡的可能机制。方法以鲎素20μg/ml处理HL-60细胞0~24 h。流式细胞仪检测线粒体膜电位。Western blot法和发色底物法检测Caspase-3、Caspase-6、Caspase-8和Caspase-9活性变化。Western blot法检测胞质中细胞色素C的变化。结果鲎素处理HL-60细胞后,线粒体膜电位下降。Caspase-3、Caspase-6、Caspase-8和Caspase-9被激活,呈时间依赖性改变。鲎素处理HL-60细胞6 h Caspase-8活性最高,而Caspase-9在12 h活性最高,Caspase-6活性在18 h达峰值,Caspase-3活性峰值时间在24 h后。胞质中细胞色素C含量呈时间依赖性增加。结论鲎素诱导HL-60细胞凋亡可能是多途径的,至少与线粒体途径和死亡受体途径相关。更多还原

【Abstract】 Objective To investigate the possible mechanism of tachyplesin inducing HL-60 cells apoptosis by studying the changes in Caspases activity.Methods HL-60 cells were treated with tachyplesin(20 μg/ml) for 0-24 h.Mitochondrial membrane potential was tested by flow cytometry.Activities of Caspases was detected by using chromogenic substrate assay and Western blot.The changes of cytochrome C in plasma of HL-60 cells were detected by Western blot.Results In HL-60 cells treated with tachyplesin,mitochondrial membrane potential was decreased;Caspases were activated in a time-dependent manner;The activity of Caspase-8 reached the peak at 6 h,Caspase-9 at 12 h,Caspase-6 at 18 h and Caspase-3 at 24 h;The level of cytochrome C in plasma of HL-60 cells was increased in a time-dependent manner.Conclusion Tachyplesin-induced apoptosis may be associated with multi-pathway,at least with the mitochondrial apoptotic pathway and death receptor pathway.更多还原