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目的研究单胺囊泡转运蛋白-2(VMAT2)抑制剂利血平(RE)作用于转染VMAT2基因的中国仓鼠卵巢细胞(VMAT2-CHO)时,VMAT2-CHO对鱼藤酮(Rotenone)、多巴胺(DA)的毒性抵抗作用。方法采用MTT比色法检测RE与不同浓度Rotenone及DA共同作用下的VMAT2-CHO细胞存活率;采用流式细胞仪检测细胞内活性氧(ROS)水平。结果Rotenone对VMAT2-CHO细胞有毒性作用,随浓度增大,细胞存活率下降;Rotenone和DA对细胞有协同毒性作用。随DA浓度增大,VMAT2-CHO细胞产生的ROS增加;VMAT2抑制剂利血平增加DA的毒性,使同样浓度的DA诱发VMAT2-CHO细胞产生的ROS增加,存活率下降。结论RE使VMAT2功能抑制时DA在胞浆内聚集,触发DA内源性毒性,通过氧化应激系统而发挥毒性作用。DA内源性毒性可协同环境毒物Rotenone增加对细胞的毒性作用。
Objective To investigate the effects of VMAT2 inhibitor reserpine (RE) on Chinese hamster ovary cells transfected with VMAT2 gene (VMAT2-CHO). The effects of VMAT2-CHO on rotenone, DA) toxicological effects. Methods MTT assay was used to detect the survival rate of VMAT2-CHO cells treated with different concentrations of Rotenone and DA. The level of reactive oxygen species (ROS) was measured by flow cytometry. Results Rotenone had toxic effect on VMAT2-CHO cells. With increasing concentration, Rotenone and DA had synergistic cytotoxic effect on cells. Increased DA concentration resulted in increased ROS production in VMAT2-CHO cells; VMAT2 inhibitor reserpine increased the toxicity of DA, resulting in increased ROS generation and decreased viability of VMAT2-CHO cells induced by DA at the same concentration. Conclusions DA inhibits the accumulation of DA in cytoplasm when VMAT2 function is inhibited, triggers the endogenous toxicity of DA and exerts toxic effects through the oxidative stress system. DA endogenous toxicity can be combined with environmental toxicants Rotenone increased toxic effects on cells.