目的：考察双氢青蒿素（dihydroartemisinin，DHA）诱导的外周T细胞淋巴瘤Hut-78细胞凋亡，并进一步研究其作用机制。方法：CCK-8法检测1~30 μg/ml DHA对Hut-78细胞增殖的影响，运用Hochest 33258染色技术在共聚集显微镜下观察DHA对Hut-78细胞核形态的影响，流式细胞术检测DHA对Hut-78细胞凋亡的影响，10 mmol/L的活性氧（reactive oxygen species，ROS）清除剂N-乙酰半胱氨酸（NAC）预处理细胞1 h，观察ROS在DHA引起的细胞线粒体膜电位变化中的作用，Western blotting检测在Hut-78细胞凋亡过程中ROS对DHA引起的细胞色素C释放的影响。结果：DHA呈浓度依赖性抑制Hut-78细胞的增殖，并引起细胞核固缩，形成凋亡小体。5、10、20 μg/ml DHA引起的细胞凋亡率分别为（25.1±2.8）%、（436±3.1）%、（68.9±2.6）%，与对照组相比差异有统计学意义（均P<0.01）。20 μg/ml DHA处理导致Hut-78细胞线粒体膜电位下降（59.4±2.6）%，而经ROS抑制剂NAC预处理之后，线粒体膜电位下降（38.4±2.1）%。DHA能够引起线粒体中细胞色素C的释放，而NAC的预处理能够明显地抑制这一过程，统计结果进一步证明这一点。结论：DHA能够抑制外周T细胞淋巴瘤Hut-78细胞的增殖并诱导细胞凋亡，其作用机制可能与DHA促进ROS依赖的细胞色素C释放有关。

Objective:To investigate dihydroartemisinin (DHA)-induced apoptosis of peripheral T lymphocytoma Hut-78 cell line and its possible mechanism. Methods: The effect of DHA 1－30 μg/ml on proliferation of Hut-78 cells was measured by CCK-8 assay. The morphological changes of Hut-78 cell's nuclei induced by DHA were observed by Hochest 33258 staining and confocal microscopy. Flow cytometry was used to examine the apoptosis of Hut-78 cells induced by DHA. After pretreating the Hut-78 cells with 10 mmol/L of reactive oxygen species (ROS) scavenger N-acetyl cysteine C (NAC) , a role of ROS in changes of mitochondrial membrane potential induced by DHA was evaluated. Effect of ROS on release of cytochondrial C induced by DHA during apoptosis of Hut-78 cells was examined with an immunoblotting assay.Results:DHA inhibited proliferation of the Hut-78 cells in a dose-dependent manner, induced nucleus pyknosis and formed apoptotic bodies in the cells. Apoptosis rates of the Hut-78 cells iuduced with 5, 10 and 20 μg/ml of DHA were (25.1±2.8)%, (43,6±3.1)% and (68.9±2.6)% respectively, which were significantly different from that of the control group (all P<0.01 ). Furthermore, treatment with 20 μg/ml of DHA droped mitochondrial membrane potential of the Hut-78 cells by (59.4±2.6)%, and pretreating with ROS scavenger NAC droped the mitochondrial membrane potential by (38.4±2.1)%. DHA can induce release of cytochrome C in mitochondrion, which can be significantly inhibited by pretreatment with NAC. Statistic results further demonstrated this point. Conclutions: DHA can effectively inhibit proliferation of the peripheral T lymphocytoma Hut-78 cells and induce their apoptosis. Its action mechanism may be related to ability of HAD to promote release of ROS-dependent cytochrome C.

河南省教育厅基金资助项目（No. 2006320039）