顺铂对大鼠肝细胞毒性及谷胱甘肽的保护作用

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48 h 3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48 h 3个时间点半数抑制浓度(IC50)分别为1.13,0.21和0.15 mmol/L;BSO能使3组IC50均降低,分别为0.017,0.011和0.013 mmol/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.

Protective effects of glutathione on cisplatin - induced toxicity to rat hepatocytes

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(IC 50) of rat hepatocytes at 8,24 and 48 hours were 1 13,0.21 and 0.15mmol/L,respectively.BSO made IC 50 of cisplatin to rat hepatocytes lower down to 0.017,0.011 and 0 013mol/L,respectively,while L-cysteine made IC 50 increase up to more than 5mmol/L.Conclusion Cisplatin has time and concentration-dependent toxicity to rat hepatocytes;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 rat hepatocytes.