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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160μmol/L的环磷酰胺干预24 h,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
OBJECTIVE: To investigate the toxic effects of cyclophosphamide on rat primary cultured hepatocytes.Methods: Rat primary hepatocytes were prepared by “two-step perfusion method” and were given 10,20,40,80 and 160μmol / L Cyclophosphamide intervention 24 h, solvent control group without cyclophosphamide intervention. Cytotoxicity of cyclophosphamide to hepatocytes was observed by MTT assay. The biochemical and intracellular oxidative and anti-oxidative indexes of supernatant in each group were determined. Results: Different concentrations of cyclophosphamide have obvious toxicity on rat primary cultured hepatocytes. Compared with the solvent control group, the levels of alanine aminotransferase and aspartate aminotransferase in the liver cell culture supernatant of each experimental group treated with cyclophosphamide were increased; the content of malondialdehyde in the cell freezing and thawing fluid increased, Superoxide dismutase and glutathione peroxidase activity were decreased (all P <0.05). Conclusion: A model of lipid peroxidation induced by cyclophosphamide in primary cultured rat hepatocytes was established successfully