双氢青蒿素对嗜热四膜虫的毒性效应

TOXIC EFFECTS OF DIHYDROARTEMISININ ON TETRAHYMENA THERMOPHILA

  • 摘要: 以嗜热四膜虫(Tetrahymena thermophila)为试验对象, 双氢青蒿素(Dihydroartemisinin, DHA)以终浓度为0(对照组)、40、80、160和320 μmol/L 分别加入到嗜热四膜虫细胞培养液中, 探讨双氢青蒿素对嗜热四膜虫的毒性作用。采用 CCK-8 法检测嗜热四膜虫细胞增殖, 倒置显微镜和荧光显微镜观察细胞的形态结构及运动, 采用流式细胞术检测线粒体膜电位, 检测细胞内抗氧化还原酶活力和线粒体酶活力。结果表明, DHA显著抑制嗜热四膜虫增殖(P<0.05), 在一定暴露时间内增殖活力和浓度呈负相关。双氢青蒿素作用嗜热四膜虫48h后各暴露组细胞皱缩变圆, 对照组细胞呈椭圆状。其中在160和320 μmol/L DHA暴露下, 嗜热四膜虫在培养基中的活动减弱, 细胞核出现固缩和浓染等特征, 线粒体膜电位显著下降(P<0.05)。随着 DHA浓度增加, 细胞内超氧化物歧化酶(SOD)、过氧化氢酶(CAT)、谷胱甘肽过氧化物酶(GSH-Px)和谷胱甘肽硫转移酶(GST)活性先增强后下降。线粒体内琥珀酸脱氢酶(SDH)活性逐渐降低, 与对照组相比, 差异均有统计学意义(P<0.05)。上述结果表明, 双氢青蒿素对嗜热四膜虫具有毒性作用, 抗氧化酶在一定程度上能抵抗双氢青蒿素暴露导致的氧化损伤。氧化应激和线粒体损伤可能是双氢青蒿素对嗜热四膜虫产生毒性效应的重要机制。

     

    Abstract: To explore the toxicity of dihydroartemisinin (DHA) to Tetrahymena thermophila, T. thermophila was exposed to DHA (0, 40, 80, 160 and 320 μmol/L), and then cell proliferation, cell structures and movement, antioxidase activity, and mitochondrial membrane potential (MMP) were measured. The results showed that DHA inhibited the growth of T. thermophila in a concentration-dependent pattern. Observation through inverted microscope showed that after 48h treatment with DHA, T. thermophila became round and shrunken compared with the cells in the control group, which were oval. Meanwhile, high concentration treatment group (160 and 320 μmol/L) reduced cell movement, induced nucleus pycnosis, and decreased MMP significantly (P<0.05). With the increase of DHA concentration, the activities of the superoxide dismutase (SOD), catalase (CAT), glutathione peroxidase (GSH-Px), and glutathione S-transferase (GST) increased at first and then decreased, and the level of succinate dehydrogenase (SDH) decreased significantly (P<0.05). The results indicated that the antioxidant enzymes in T. thermophila could prevent the oxidative damage caused by DHA exposure. Based on our results, the toxic effects of DHA on T. thermophila might be related with oxidative damage and mitochondrial damage.

     

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