环磷酰胺对大鼠肝细胞的毒性

目的：探讨环磷酰胺对大鼠原代培养肝细胞的毒性作用.方法：采用“二步灌流法”制备大鼠原代肝细胞,分别给予10、20、40、80及160 μmol/L的环磷酰胺干预24h,溶剂对照组不予环磷酰胺干预.噻唑蓝（ MTT）法观察环磷酰胺对肝细胞的毒性,测定各组培养基上清液生化和细胞内氧化及抗氧化指标.结果：不同浓度的环磷酰胺对大鼠原代培养肝细胞均具有明显的毒性.与溶剂对照组比较,环磷酰胺干预的各实验组肝细胞培养上清液丙氨酸氨基转移酶、天冬氨酸氨基转移酶均升高；细胞冻融液丙二醛含量均升高,超氧化物歧化酶及谷胱甘肽过氧化物酶活性均降低（均P ＜0.05）.结论：成功建立了环磷酰胺致大鼠原代培养肝细胞脂质过氧化模型.

The hepato-toxicity of cyclophosphamide in rats

Objective：To study the toxicity of cyclophosphamide on primary culture of rat hepatoeytes. Methods： The primary culture of rat hepatocytes was prepared using two-step perfnsion method, and then subjected to 24 h incubation with 10 μmol/L, 20 μmoL/L, 40 μmol/L, 80μmol/L or 160μmol/L cyclophosphamide. A control group of hepatocytes was incubated with solvent alone. MTT assay was used to determine the cytotoxicity of cyclophosphamide. The biochemical parameters of cell supernatant, intracellular oxidatiorr/antioxidation in each group were also measured. Results： All doses of Cyclophosphamide showed significant toxicity on primarily cultured rat hepatocytes. Compared with the controls, the contents of alanine aminotransferase and aspartic transaminase in supernatant of hepatocytes were increased in all groups treated with Cyclophosphamide. In addition, significantly higher level of malonaldehyde and lower levels of superoxide dismutase and glutathione peroxidase were found in freezing and thawing medium from experiment groups compared with the control group （ P 〈 O. 05 ）. Conclusion ： The cyclophosphamide-induced lipid peroxidation model of primarily cultured rat hepatocytes was successfully established.