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目的 :探讨异环磷酰胺 (Ifo)对混悬培养大鼠肝细胞的毒性效应及其可能机制。方法 :以两步灌流法消化成年大鼠肝细胞 ,并进行混悬培养。Ifo以 5,10 ,2 0mmol/L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、丙二醛 (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 ,培养液中乳酸脱氢酶 (LDH)、天冬氨酸氨基转移酶 (AST)活性增高 ,同时肝细胞总巯基 (TSH)、非蛋白巯基 (NPSH)、蛋白巯基 (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Ifo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 ,滑面内质网扩张 ,呈囊泡状改变。结论 :Ifo对混悬培养大鼠肝细胞有损伤作用 ,巯基物质的降低在Ifo肝细胞毒性中起重要作用
Objective: To investigate the toxic effects of Ifo on cultured rat hepatocytes and its possible mechanism. Methods: Adult rat hepatocytes were digested by two-step perfusion method and cultured in suspension. Ifo treated with 5, 10, 20 mmol / L, the survival rate of hepatocytes, the leakage of intracellular enzymes and the state of hepatocyte thiol, the content of malondialdehyde (MDA) in hepatocytes were observed 3 h after exposure. Morphology and ultrastructure were observed. Results: With the increase of exposure concentration, the survival rate of hepatocytes decreased gradually and the intracellular enzyme leakage increased. The activities of lactate dehydrogenase (LDH) and aspartate aminotransferase (AST) in the culture fluid increased. TSH, NPSH and PSH decreased gradually, and the decrease of PSH played a major role in TSH depletion. No significant increase in MDA content in liver cells was observed. Morphological examination showed that Ifo hepatoblasts appeared on the surface of the “big blisters”, intracellular mitochondria swelling, vacuolization, rough endoplasmic reticulum dilation, partial degranulation, lumen obstruction, glomerular endoplasmic reticulum dilated, was vesicular change. CONCLUSION: Ifo can damage rat hepatocytes cultured in suspension, the decrease of sulfhydryl group plays an important role in the toxicity of Ifo hepatocytes