微囊藻毒素-LR对中国仓鼠卵巢细胞中过氧化氢酶活力和丙二醛含量的影响

Effects of Microcystin-LR on CAT and MDA in Chinese Hamster Ovary Cells

目的通过检测中国仓鼠卵巢(CHO)细胞中过氧化氢酶(CAT)活力和丙二醛(MDA)含量的变化研究微囊藻毒素-LR(MC-LR)对CHO细胞的氧化应激作用。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MTT法测细胞活性,计算半致死效应浓度(EC50)。调整细胞密度约为1×106/ml,分别用终浓度为0(对照)、2.5(1/4 EC50)、5(1/2 EC50)、10μg/m(lEC50)的MC-LR染毒24 h后,测定细胞中CAT活力和MDA含量。结果随着MC-LR染毒浓度的升高,CHO细胞活性呈下降趋势。与对照组相比,1μg/ml染毒组细胞活性降低,但差异无统计学意义;5、10、15μg/ml MC-LR染毒组CHO细胞活性较低,差异均有统计学意义(P<0.05),且10μg/ml MC-LR染毒组细胞活性为53.8%,接近50%,故EC50约为10μg/ml。与对照组相比,各浓度MC-LR染毒组CHO细胞内CAT活力较低,MDA含量较高,差异均有统计学意义(P<0.05)。结论 MC-LR染毒可使CHO细胞中CAT活力降低,MDA含量升高,提示其对CHO细胞有氧化损伤作用。

Objective To study the oxidative stress of Chinese hamster ovary （CHO） cells induced by microcystins-LR through testing the changes of catalase （CAT） and malondialdehyde （MDA）. Methods CHO Cells were seeded at lxlOs cells per well of a 96 well-plate for 24 h. The cells were exposed to different doses of microcystin-LR （0, 1, 5, 10 and 15 μg/ml） for 24 h. The viability of ceils was determined with MTT assay. Meanwhile,the ECru value was determined. The cells were inoculated at 1×10^6 cells per well of a 6 well-plate and were exposed to different doses of microcystin-LR [0 μg/ml,2.5 μg/ml （1/4 EC50）,5 μg/ml （1/2 EC50）, 10 μg/ml （EC50）] for 24 h. Then,viability of CAT and the content of MDA were determined. Results With the increases of MC-LR concentration,CHO cell viability showed a declining curve. Compared with the control group,a significant reduce of cytoactive were seen in 5, 10 and 15 μg/ml groups （P〈0.05）. The cytoactive in 10 μg/ml group was 53.8% which closed to 50% ,so EC50 was about 10 μg/ml. Compared with the control group,the viability of CAT in every MC-LR exposed group was significantly reduced （P〈0.05）,and the content of MDA was significantly increased （P〈0.05）. Conclusion The results in the present study suggest that MC-LR may cause oxidative damage on CHO cells.