Department of Pharmacology, School of Pharmaceutical Sciences, Jilin University, and Department of Infectious Diseases, First Hospital, Changchun, PR China.
Food Chem Toxicol. 2012 Mar;50(3-4):590-6. doi: 10.1016/j.fct.2012.01.002. Epub 2012 Jan 14.
Juglone, a major chemical constituent of Juglans mandshruica Maxim, is a promising anticancer agent that has shown a strong activity against cancer cells in vitro. Our previous study showed that juglone inhibited the proliferation of HL-60 cells with an IC50 value ∼8 μM. To further explore the proapoptotic mechanism of juglone, we investigated the role of the reactive oxygen species (ROS) in the apoptosis induced by juglone in HL-60 cells. The generation of ROS was about 2 to 8-fold as compared to control cell after treatment with juglone (2, 4 and 8 μM) for 24 h. The glutathione (GSH) depletion was consistent with ROS generation after treatment with juglone. Reversal of apoptosis in antioxidants (NAC and catalase) pretreated cells indicated the involvement of ROS in juglone-induced apoptosis. The cleavage of PARP and procaspase-3 and -9, loss of mitochondrial membrane potential (△Ψm), and release of cytochrome c (Cyt c) and Smac induced by juglone were significantly blocked by NAC. NAC also prevented the inhibition the phosphorylation of Akt and mTOR proteins by juglone. Collectively, these results indicated that ROS played a significant role in the apoptosis induced by juglone in human leukemia cell HL-60.
胡桃醌是胡桃属植物的主要化学成分之一，是一种很有前途的抗癌药物，在体外对癌细胞具有很强的活性。我们之前的研究表明，胡桃醌抑制 HL-60 细胞的增殖，IC50 值约为 8 μM。为了进一步探讨胡桃醌诱导细胞凋亡的促凋亡机制，我们研究了活性氧（ROS）在胡桃醌诱导 HL-60 细胞凋亡中的作用。与对照组相比，用胡桃醌（2、4 和 8 μM）处理 24 小时后，ROS 的生成增加了 2 到 8 倍。用胡桃醌处理后，谷胱甘肽（GSH）的耗竭与 ROS 的产生一致。抗氧化剂（NAC 和过氧化氢酶）预处理细胞中的凋亡逆转表明 ROS 参与了胡桃醌诱导的凋亡。NAC 显著阻断了胡桃醌诱导的 PARP 和 procaspase-3 和 -9 的裂解、线粒体膜电位（△Ψm）的丧失、细胞色素 c（Cyt c）和 Smac 的释放。NAC 还阻止了胡桃醌对 Akt 和 mTOR 蛋白磷酸化的抑制。总之，这些结果表明 ROS 在胡桃醌诱导的人白血病细胞 HL-60 凋亡中起重要作用。
