环磷酰胺对离体大鼠肝细胞毒作用机理研究  

Study on Toxicity Mechanism(s) of Cyclophosphamide in Cultured Suspensibly Rat Hepatocytes

作　　者：施畅[1] 廖明阳[1] 郭巧珍[1] 盛和章[1] 

机构地区：[1]中国人民解放军军事医学科学院毒物药物研究所,北京100850

出　　处：《解放军药学学报》2001年第3期128-131,共4页Pharmaceutical Journal of Chinese People's Liberation Army

摘　　要：目的　探讨环磷酰胺 (Cp)对混悬培养大鼠肝细胞的毒性效应及其可能机理。方法　以两步灌流法消化成年大鼠肝细胞 ,并进行混悬培养。Cp以 5、10、2 0mmol·L-1染毒 ,观察染毒后 3h肝细胞的存活率、胞内酶泄漏情况以及肝细胞巯基状态、MDA含量的变化 ,并对肝细胞表面形态和超微结构进行观察。结果　随着染毒浓度的增大 ,肝细胞存活率逐渐下降 ,胞内酶泄漏加重 ,培养液中LDH、ALT、AST活性增高 ,同时 ,肝细胞TSH、NPSH、PSH也逐渐下降 ,其中PSH下降在TSH耗竭中起主要作用。肝细胞MDA含量未发现有显著增高。形态学检查发现Cp使肝细胞表面出现“大疱” ,胞内线粒体肿胀 ,空泡化 ,粗面内质网扩张 ,部分脱颗粒 ,内腔模糊 ,核固缩 ,核内染色质边集成块状。结论　Cp对混悬培养大鼠肝细胞有损伤作用 。Aim To study the toxicity mechanism(s) of Cyclophosphamide (Cp) on suspensibly cultured rat hepatocytes.Methods Hepatocytes of adult rat were isolated using two-step perfusion method and cultured suspensibly. Cell viability、intracellular enzymes leakage、 contents of sulfhydryl groups and MDA contents of hepatocytes were detected 3 hours after Cp was administered at 5、10、20mmol·L -1 .Surface and ultrastructure of hepatocytes were also observed.Results Cell viability and TSH、NPSH、PSH contents of hepatocytes significantly declined, and LDH、ALT、AST activities in media increased due to the leakage of intracellular enzymes.The decrease of PSH content was ascribed to TSH's depletion. The higher the dose was, the more serious these changes became. However, MDA contents of the hepatocytes were not found increased at any doses of Cp treated groups. In pathological examination,“bulla'formation was found on the surface of the hepatocytes, deformation、swelling even vacuolation of mitochondria and dilation of rough endoplasmic reticulum were also observed. Furthermore, the nucleuses of injured hepatocytes were found shrunk and chromatin congregated to the edge of the nucleus membrane.Conclusions Cp has toxic effects on suspensibly cultured rat hepatocytes.The decrease of sulfhydryl groups contributes to the hepatotoxicity induced by Cp.

关 键 词：环磷酰胺 肝细胞 巯基 毒性作用 

分 类 号：R979.1[医药卫生—药品] R961[医药卫生—药学]