贝母素甲对多药耐药人白血病细胞活力和凋亡的影响及机制

摘要

目的 观察贝母素甲对多药耐药白血病人细胞K562/A02活力与细胞凋亡的影响,探讨活性氧(ROS)在此过程中的作用.方法 将K562/A02细胞成3组,药物组分别加入终浓度为100、200、400 μmol/L的贝母素甲;NAC预处理组采用5 mmlo/L ROS清除剂N-乙酰-L-半胱氨酸(NAC)预处理后,再加入终浓度为400 μmol/L的贝母素甲;对照组加等体积的药物溶解介质RPMI1640培养基.采用MTT法检测各组细胞活力,流式细胞术检测细胞凋亡情况,紫外分光光度法检测细胞内ROS、GSH水平.结果 与对照组比较,药物组细胞活力均降低(P<0.05或<0.01)、细胞内ROS水平增高(P均<0.01)、GSH水平下降(P均<0.01)、细胞凋亡率上升(P均<0.05),NAC预处理组的细胞活力、细胞凋亡率和ROS水平无明显变化(P均>0.05).结论 贝母素甲可抑制多药耐药人白血病细胞K562/A02的细胞活力、诱导其凋亡;其机制是通过诱导细胞ROS爆发,引起GSH水平下降,从而逆转多药耐药的发生.%Objective To investigate the effects of peimine on cell viability and apoptosis of multidrug resistant human leukemia K562/A02 cells and the function of reactive oxygen species (ROS) in this progress.Methods The K562/A02 cells were divided into three groups.Cells in the drug group were treated with peimine with the final concentrations of 100, 200, and 400 μmol/L, respectively.Cells in the N acetyl cysteine (NAC) pretreatment group were treated with 5 mmol/L antioxidant NAC, followed by 400μmol/L peimine.Cells in the control group were treated with an equal volume of dissolution medium RPMI1640.The cell viability of each group was detected by MTT, the apoptosis was detected by flow cytometry, and the levels of ROS and glutathione (GSH) in the cells were detected by UV spectrophotometry.Results Compared with the control group, the cell viability of the drug group decreased (P<0.05 or P<0.01), intracellular ROS level increased (P<0.01), GSH concentration decreased (P<0.01), and the apoptosis rate increased (P<0.05).However, the cell viability, the apoptosis rate, and the ROS level in NAC pretreatment group showed no obvious change (all P>0.05).Conclusion Peimine inhibits the cell viability and induces apoptosis of multidrug resistant leukemia K562/A02 cells through inducing the ROS outbreak and thus increasing the GSH content.