顺铂对大鼠肝细胞毒性及谷胱甘肽的保护作用
Protective effects of glutathione on cisplatin-induced toxicity to rat hepatocytes
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摘要:目的 探讨顺铂对原代培养的大鼠肝细胞的毒性及谷胱甘肽对其影响。方法 从大鼠的肝脏分离培养肝实质细胞接种于96孔培养板, 培养3h后加入一系列浓度的顺铂, 或在加入顺铂前16和4h, 分别加入谷胱甘肽(glutathione, GSH)合成抑制剂DL-buthionine-(S, R)-sulfoximine(BSO)和GSH的前体物半胱氨酸, 继续培养, 分别在8, 24和48h3个时间点用噻唑蓝(MTT)方法检测细胞存活率。结果 顺铂对大鼠肝细胞有明显的毒性, 在不同的时间点有各自的剂量-反应关系, 顺铂对大鼠肝细胞在8, 24和48h3个时间点半数抑制浓度(IC50)分别为1.13, 0.21和0.15mmol/L; BSO能使3组IC50均降低, 分别为0.017, 0.011和0.013mmol/L, 而半胱氨酸则可使3组IC50均升高, 均大于5mmol/L。结论 顺铂对大鼠肝细胞具有明显的毒性作用, 且呈时间和剂量依赖关系; BSO可增强顺铂的肝细胞毒性, 而半胱氨酸对顺铂引起的肝细胞毒性有保护作用, 提示细胞内GSH对顺铂所致肝细胞毒性有保护作用。Abstract:Objective To investigate the effects of glutathione on cisplatin-induced toxicity to rat hepatocytes.Methods Hepatocytes were isolated from rats and then inoculated into 96 well plates, after pretreatment of L-cysteine(precursor of GSH synthesis)or BSO(inhibitor of GSH synthesis)for 4 hours and 16 hours, respsectively.A series of concentrations of cisplatin, in the presence and absence of L-cysteine, were added and coincubated, MTT assays were performed to test cell viability at 8, 24 or 48 hours after coincubation.Results Cisplatin had time-dependent and concentration-dependent cytotoxicity.The concentrations of cisplatin that inhibited 50% cell growth(IC50)of rathepatocytes at 8, 24 and 48 hours were 1.13, 0.21 and 0.15 mmol/L, respectively.BSO made IC50 of cisplatin to rathepatocytes lower down to 0.017, 0.011 and 0.013 mol/L, respectively, while L-cysteine made IC50 increase up to more than 5 mmol/L.Conclusion Cisplatin has time and concentration-dependent toxicity to rathepatocytes; BSO can enhance cisplatin-induced cytotoxicity, while L-cysteine can protect cisplatin-induced cytotoxicity.It was possible that GSH can protect cisplatin-induced toxicity to rathepatocytes.
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