摘要
用活体肝脏灌注、胶原酶消化法分离大鼠肝细胞并作原代培养 ,以 Cd Cl2 ( 0～ 2 50或 50 0 μmol/ L)处理 ,测定培养上清液中 LDH、ALT、ARG和细胞内 GSH-Px、GSH、MDA等的含量 ,探讨镉对肝细胞的损伤作用和脂质过氧化状况。结果 ,当 Cd≥ 1 0和 5μmol/ L时 ,细胞内 LDH、ARG泄漏分别明显增加 ,GSH-Px活力显著受到抑制 ,GSH含量和 MDA产生随着镉的剂量增大而增加 ( Cd≥ 50μmol/ L)。认为镉对肝细胞的损伤和对细胞膜完整性的破坏 ,与脂质过氧化有关 ;镉处理后细胞中 GSH增加可能与镉抑制了还原型谷胱甘肽向氧化型谷胱甘肽的转化有关。镉可直接抑制 ALT本身的活性 ,因而认为以细胞内
Rat hepatocytes were isolated by a collagenase perfusion in situ method and the primary hepatocyte cultures were used to evaluate the cadmium induced cellular injury,intracellular enzyme leakage and lipid peroxidation(LPO).Isolated hepatocytes were incubated at 37℃ with cadmium chloride(0 250 or 500 μmol/L).Activities of lactate dehydrogenase(LDH),alanine aminotransferase(ALT),arginase(ARG) and glutathione peroxidase(GSH Px) in cell free medium were determined at 6 h interval.At the end of incubation,cellular glutathione(GSH) and malondialdehyde(MDA) content and GSH Px activity were measured.The results obtained showed that incubation of hepatocytes with Cd(≥10 and 5 μmol/L respectively) resulted in a significant increase in enzyme leakage of LDH and ARG.Cadmium concentrations of higher than 5 μmol/L decreased GSH Px activity.Intracellular GSH content and MDA production increased with the addition of Cd(≥50 μmol/L).The data indicated that cadmium could induce cellular injury and damage the plasma membrane integrity,and resulted in cadmium induced lipid peroxidation.It is speculated that the increased GSH level in Cd treated hepatocytes may result from the inhibition of the conversion of GSH into GSSG.In addition,cadmium may inhibite the ALT activity itself,so that it should be with caution when ALT measurement was used as monitoring enzyme leakage induced by cadmium.
出处
《中国兽医学报》
CAS
CSCD
北大核心
2000年第6期580-583,共4页
Chinese Journal of Veterinary Science
基金
国家自然科学基金!资助项目 ( 39870 5 6 7)
江苏省自然科学基金!资助项目 ( BK970 72 )
