活性氧在δ-榄香烯诱导人结肠腺癌DLD-1细胞凋亡中的作用研究  

The role of ROS in apoptosis induced by δ-elemene in colorectal adenoarcinoma cells

作　　者：王茜莎[1] 杨威 陶淑娟[3] 董金华[3] 潘雪刁[1] 张德志[1] 

机构地区：[1]广东药学院药科学院,广州510240 [2]广州医药工业研究院药物非临床评价中心 [3]沈阳药科大学制药工程学院

出　　处：《环球中医药》2010年第4期253-257,共5页Global Traditional Chinese Medicine

基　　金：辽宁省教育厅基金项目(202043228)

摘　　要：目的探讨δ-榄香烯诱导人结肠腺癌DLD-1细胞凋亡的机制以及活性氧(reactive oxygen species,ROS)在δ-榄香烯诱导人结肠腺癌细胞DLD-1凋亡中的作用。方法采用MTT法考察了δ-榄香烯对人结肠腺癌DLD-1细胞和人正常肝胚胎WRL-68细胞增殖的影响;采用氯甲基二氯荧光素二乙酸酯(CM-H2DCFDA)定量检测凋亡过程中ROS的变化;流式细胞术JC-1标记检测凋亡细胞中线粒体膜电位(Δφm)的影响。结果δ-榄香烯明显抑制DLD-1细胞的增殖,对WRL-68细胞的细胞毒作用不明显。200μM的δ-榄香烯作用DLD-1细胞1小时、3小时和6小时所引起的ROS含量的增加,分别接近空白对照组的7.8倍、18.2倍和21.8倍;这种作用可以被谷胱甘肽(glu-tathione,GSH)(ROS的清除剂)显著性抑制,即加入GSH后δ-榄香烯对ROS生成率(3.46%)与空白对照(2.11%)相比无显著性差异。流式细胞仪检测结果表明,线粒体膜电位(Dφm)的降低呈时间依赖性,6小时、12小时、24小时线粒体膜电位降低量分别是空白组的(2.24±0.12)倍,(2.85±0.33)倍,(5.11±0.24)倍。结论Δφm的降低表明δ-榄香烯的抗肿瘤作用是以诱导肿瘤细胞凋亡的方式产生的,且其凋亡过程提示可能与ROS的生成增加相关。Objective To investigate the apoptosis effect and the role of ROS in the mechanism of δ-elemene on Colorectal Adenoarcinoma Cells DLD-1 in vitro. Methods Cell proliferation was measured by MTT method. Apoptosis was further confirmed and quantified by the mitoehondrial probe JC-1 using flow cytometry. The rapid increase in intracellular reactive oxygen species （ROS） levels was involved in the mechanism of cell death. Results δ-elemene induced DLD-1 cell apoptosis and a weaker cytotoxic effect on WRL-68 cell. There was approximately 7. 8, 18.2 or 21.8 fold increase in generation of ROS in 200 μM δ-elemene for l h, 3h or 6h compared with vector control cells, the increase was statistically significanL Furthermore, The change of ROS induced by δ-elemene was inhibited by GSH to a level （3.46%） that was almost as low as the untreated control （2. 11% ）. the trends of increasing incidences of compromised △φm for 6h, 12h or 24h were （2. 24±0. 12）, （2. 85±0. 33 ）, （5.11±0. 24） fold with respected to the untreated control. Conclusions These results indicate that apoptosis pathway in DLD-1 cells by δ--elemene was associated with the ROS generation.

关 键 词：δ-榄香烯 人结肠腺癌DLD-1细胞 凋亡 活性氧 线粒体膜电位 

分 类 号：R285[医药卫生—中药学]