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目的:利用原代培养正常大鼠肝细胞研究苯巴比妥钠(PNB)对环磷酰胺(CP)致离体大鼠肝细胞的毒性作用。方法:用两步灌注法分离原代大鼠肝细胞;观察苯巴比妥钠(PNB)对环磷酰胺(CP)致离体大鼠肝细胞的毒性作用。采用四甲基噻唑蓝(MTT)法测定PNB对原代大鼠肝细胞的最大无毒浓度(TC0),先用PNB的最大无毒浓度(TC0)诱导原代大鼠肝细胞24h,再直接加含不同浓度CP的培养液培养24h,MTT法测药物对细胞生长抑制率的影响,计算IC50。并测定药物作用后培养液上清中AST、ALT和LDH活性及培养液中GSH和细胞内GSH的含量。结果:经苯巴比妥钠诱导后的环磷酰胺各剂量组与未经苯巴比妥钠诱导的环磷酰胺各剂量组比较,细胞生长抑制率降低,培养液上清AST、ALT和LDH活性降低,培养液上清和细胞内GSH含量升高。结论:苯巴比妥钠能减轻环磷酰胺致离体大鼠肝细胞的毒性作用。
Objective: To study the toxic effects of phenobarbital sodium (PNB) on hepatocytes induced by cyclophosphamide (CP) in rat primary hepatocytes. Methods: The primary rat hepatocytes were isolated by two-step perfusion method. The toxic effects of phenobarbital sodium (PNB) on hepatocytes induced by cyclophosphamide (CP) were observed. The maximum non-toxic concentration (TC0) of PNB on primary rat hepatocytes was determined by MTT method. The primary rat hepatocytes were induced with maximum non-toxic concentration of PNB (TC0) for 24 h and then directly The medium containing different concentrations of CP was cultured for 24 hours. The effect of the drugs on the growth inhibition rate was measured by MTT method and the IC50 was calculated. The activities of AST, ALT and LDH in culture supernatant and the content of GSH and GSH in the culture medium were also determined. Results: Compared with the cyclophosphamide-induced dose groups of cyclophosphamide induced by phenobarbital sodium, the cell growth inhibition rates decreased, and the levels of AST, ALT and LDH Activity decreased, medium supernatant and intracellular GSH content increased. Conclusion: Phenobarbital sodium can reduce the toxic effects of cyclophosphamide on isolated rat hepatocytes.