Innes G K, Fuller B J, Hobbs K E
Academic Department of Surgery, Royal Free Hospital School of Medicine, London, UK.
In Vitro Cell Dev Biol. 1988 Feb;24(2):126-32. doi: 10.1007/BF02623889.
Rat hepatocytes were isolated and then maintained in serum-free cell culture medium for 24 h. The amount of malondialdehyde (MDA) accumulated in the medium was assayed and used as a measure of lipid peroxidation. The activity of lactate dehydrogenase (LDH) and urea were measured in the medium and used as indicators of hepatocellular viability and function. The effects of iron; desferrioxamine mesylate (Desferal), an iron chelator; and mannitol, a hydroxyl free radical scavenger were investigated. The addition of iron, Fe2 resulted in a three-fold increase in the levels of MDA. Desferal inhibited the production of MDA and blocked the effect of Fe2+. Neither iron nor Desferal had any effect on LDH or urea levels. Mannitol had no effect on MDA or urea production, but caused a 4 to 8-fold increase in the LDH levels in the medium. The results show that iron is involved in the mechanism of lipid peroxidation in hepatocyte cultures but suggest that as a pathologic event lipid peroxidation is not expressed in terms of viability during the first 24 h of hepatocyte culture.
分离大鼠肝细胞,然后在无血清细胞培养基中培养24小时。检测培养基中积累的丙二醛(MDA)量,并将其用作脂质过氧化的指标。测定培养基中乳酸脱氢酶(LDH)活性和尿素含量,并将其用作肝细胞活力和功能的指标。研究了铁、铁螯合剂甲磺酸去铁胺(去铁敏)和羟基自由基清除剂甘露醇的作用。添加铁离子(Fe2+)导致MDA水平增加了三倍。去铁敏抑制MDA的产生并阻断Fe2+的作用。铁和去铁敏对LDH或尿素水平均无任何影响。甘露醇对MDA或尿素产生没有影响,但导致培养基中LDH水平增加4至8倍。结果表明,铁参与肝细胞培养中脂质过氧化的机制,但表明作为一种病理事件,脂质过氧化在肝细胞培养的最初24小时内并未以活力的形式表现出来。