摘要
目的探讨靛玉红衍生物——PHⅡ-7对K562和K562/A02的体外杀伤作用及其机制。方法利用WST-8试剂盒、细胞凋亡检测以及活性氧(ROS)检测研究PHⅡ-7对K562和K562/A02发生杀伤作用的机制。共聚焦显微镜及Western blot分析观察药物处理前后细胞的变化。结果细胞毒实验证实PHⅡ-7对K562和K562/A02有相近的杀伤作用。给予上述两种细胞2μmol·L-1PHⅡ-7可见明显的ROS水平升高。PHⅡ-7对K562及K562/A02具有诱导凋亡的作用,呈剂量依赖性。加入ROS的抑制剂NAC可抑制给药后细胞内ROS水平的提高,同时也抑制了PHⅡ-7处理后细胞的凋亡。共聚焦显微镜观察发现药物处理的K562和K562/A02细胞出现明显的凋亡形态改变,Western blot分析表明单用PHⅡ-7可以引起PARP-1及caspase-3,caspase-9的切割,当PHⅡ-7与NAC共同作用之后,上述改变均被抑制。结论 PHⅡ-7可以通过诱导K562/和K562/A02凋亡来实现对上述细胞的杀伤作用,该作用很有可能与其升高细胞内的ROS水平有关。
Aim To investigate the effect of PHⅡ-7 on the cytotoxic effects of K562 and K562/A02 and its related mechanism.Methods Cell viability was detected by WST-8 assay;apoptosis analysis was performed by apoptosis detection kit and flow cytometry;ROS detection was also completed by flow cytometry.Confocal microscope and Western blot analysis was used to detect the changes after treatment with PHⅡ-7.Results PHⅡ-7 had almost equally cytotoxic effect on K562 and K562/A02 cell lines.Elevated ROS production was observed in both the two cell lines after treatment with PHⅡ-7.PHⅡ-7 could induce apoptosis on K562 and K562/A02,and the apoptotic rate was significantly reduced when coincubated with 5 mmol·L-1 NAC.Confocal microscope observation found conspicuous morphological changes after treatment of PHⅡ-7,furthermore,Western bolt analysis confirmed that coincubation with NAC could greatly attenuate the cleavage of PARP-1,caspase-3 and caspase-9 induced by PHⅡ-7.Conclusion PHⅡ-7 can achieve its cytotoxic effect on K562 and K562/A02 cells by apoptosis induction,and this effect is likely performed by increasing ROS production within the cells.
出处
《中国药理学通报》
CAS
CSCD
北大核心
2012年第7期911-916,共6页
Chinese Pharmacological Bulletin
基金
国家自然科学基金资助项目(No 30873091,30971291)
