目的 研究二烯丙基二硫(DADS)诱导人白血病K562细胞凋亡的分子机制.方法 应用MTT法检测细胞的活性;用流式细胞术检测细胞凋亡以及细胞内的活性氧(reactive oxygen species,ROS)水平;Western blot检测JNK以及磷酸化JNK的活化.结果 DADS能明显抑制K562细胞的增殖,呈时间和剂量依赖性; 5.0 mg·L-1DADS处理K562细胞,细胞内ROS水平在1 h后明显增加,8 h达到高峰,随后又开始下降.随着DADS剂量的增加,JNK的活性明显增强,在DADS处理K562细胞8 h后,磷酸化的JNK达到最高值,而在随后的4 h又明显降低.Sp600125和NAC能明显减少磷酸化JNK的表达和抑制DADS诱导的细胞凋亡.结论 ROS是DADS诱导K562 细胞凋亡过程中JNK活化的有效调节剂,DADS通过ROS介导的JNK活化诱导人白血病K562细胞凋亡.%Aim To research the molecular mechanisms of DADS-induced apoptosis of human leukemia K562 cells.Methods Cell viability was measured by MTT; cell apoptosis was tested by flow cytometer; Levels of DADS-induced ROS were measured by 2',7'-di-chlorofluorescein diacetate ( DCFH-DA ) fluorescence; DADS-induced phosphorylated JNK levels were measured by western blot.Results The DADS-treated K562 cells showed a dose- and time-dependent decrease in cell viability and proliferation.Levels of DADS-induced ROS also showed time-dependent increases in K562 cells.K562 cells exposed to 5.0 mg ·L-1 DADS for 8 h showed maximum levels of phosphorylated JNK, which decreased when exposed for additional 4 hours.In contrast, Sp600125, a specific inhibitor of JNK, blocked apoptosis of K562 cells exposed to DADS.N-acetylcysteine ( NAC ), a known antioxidant, also decreased ROS generation, effectively blocked apoptosis, and decreased DADS-induced phosphorylated JNK levels.Conclusion JNK is involved in DADS-induced ROS-mediated apoptosis of K562 cells.
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