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目的研究微囊藻毒素-LR(MC-LR)致中国仓鼠卵巢(CHO)细胞的凋亡作用。方法调整CHO细胞密度约为1×105/ml,分别用终浓度为0(对照)、1、5、10、15μg/ml的MC-LR染毒24 h后,采用噻唑兰(MTT),法测细胞活性,计算半致死效应浓度(EC50)。分别用终浓度为0(对照)、2.5μg/m(l1/4 EC50)、5μg/m(l 1/2 EC50)、10μg/m(lEC50)的MC-LR染毒24 h后,测定活性氧(ROS)含量、线粒体膜电位、天冬氨酸特异性半胱氨酸蛋白酶-3(caspase-3)的活力和细胞凋亡率。结果与对照组相比,各浓度MC-LR染毒组CHO细胞内ROS含量、caspase-3活力和细胞凋亡率均较高,线粒体膜电位降低,差异均有统计学意义(P<0.05);且随着MC-LR染毒浓度的升高,CHO细胞内ROS含量、caspase-3活力和细胞凋亡率均呈上升趋势,线粒体膜电位呈下降趋势。结论 MC-LR暴露可诱导体外培养的CHO细胞发生凋亡。
Objective To study the apoptosis of Chinese hamster ovary (CHO) cells induced by microcystin-LR (MC-LR). Methods The density of CHO cells was adjusted to about 1 × 10 5 / ml. The cells were treated with MC-LR at concentrations of 0, 1, 5, 10 and 15 μg / ml for 24 h, respectively. MTT Cell viability was measured and the lethal concentration (EC50) calculated. The activity was determined after 24 h exposure to MC-LR at a final concentration of 0 (control), 2.5 μg / m (l / 4 EC50), 5 μg / m l 1/2 EC50, and 10 μg / (ROS) content, mitochondrial membrane potential, caspase-3 activity and apoptosis rate. Results Compared with the control group, the ROS content, the activity of caspase-3 and the apoptosis rate of CHO cells in MC-LR treated groups were all higher than those in the control group, and the mitochondrial membrane potential was decreased (P <0.05) The ROS content, caspase-3 activity and apoptosis rate in CHO cells increased with the increase of MC-LR concentration, and the mitochondrial membrane potential decreased. Conclusion MC-LR exposure can induce apoptosis of CHO cells cultured in vitro.