4-hydroxynonenal and malondialdehyde hepatic protein adducts in rats treated with carbon tetrachloride: Immunochemical detection and lobular localization
4-hydroxynonenal and malondialdehyde hepatic protein adducts in rats treated with carbon tetrachloride: Immunochemical detection and lobular localization
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DOI：
10.1006/taap.1999.8788
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发表时间：
1999-11-15
影响因子：
3.8
通讯作者：
Petersen, DR
中科院分区：
文献类型：
--
作者：
Hartley, DP;Kolaja, KL;Petersen, DR
CCl 4的代谢引发多不饱和脂肪酸的过氧化，产生α，β-不饱和醛，如4-羟基壬烯醛（4-HNE）和丙二醛（MDA）。这些亲电性化合物的易反应性表明它们在诸如CCl 4的化合物的毒性中起作用。为了确定CCl 4引发的脂质过氧化导致4-HNE和/或MDA肝蛋白加合物形成的速率，对大鼠胃内给予CCl 4（1.0 ml/kg），并在给药后0-72 h处以安乐死。针对4-HNE-或MDA-蛋白表位的兔多克隆抗血清用于免疫组织化学和免疫沉淀/Western分析，以检测石蜡包埋的肝切片和肝匀浆中的4-HNE和MDA-蛋白加合物。早在6小时后CCl 4暴露，4-HNE和MDA加合物的免疫组化检测肝细胞定位到2区的肝腺泡。随着脂质过氧化和肝细胞坏死的增加，肝损伤进行性至24小时。在CCl 4给药后24 h观察到CCl 4肝毒性的标志，3区坏死，在2区和3区观察到免疫阳性肝细胞。CCl 4给药后36至72 h，免疫阳性细胞不再可见。从6至48小时后，CCl 4管理，至少有四个加合物蛋白免疫沉淀与抗MDA或抗4 HNE血清，这对应于80，150，205，大于205 kDa的分子量的肝匀浆。这些结果表明，4-HNE和MDA烷基化特定的肝蛋白在一个时间依赖性的方式，这似乎是与肝细胞损伤后，四氯化碳暴露。(C)北京：科学出版社.
The metabolism of CCl4 initiates the peroxidation of polyunsaturated fatty acids producing alpha,beta-unsaturated aldehydes, such as 4-hydroxynonenal (4-HNE) and malondialdehyde (MDA). The facile reactivity of these electrophilic aldehydic products suggests they play a role in the toxicity of compounds like CCl4. To determine the rate at which CCl4-initiated lipid peroxidation results in the formation of 4-HNE and/or MDA hepatic protein adducts, rats were given an intragastric dose of CCl4 (1.0 ml/kg) and euthanized 0-72 h after administration. Rabbit polyclonal antisera directed toward 4-HNE- or MDA-protein epitopes were employed in immuno-histochemical and immuno-precipitation/Western analyses to detect 4-HNE and MDA-protein adducts in paraffin-embedded liver sections and liver homogenates. As early as 6 h post CCl4 exposure, 4-HNE and MDA adducts were detected immuno-histochemically in hepatocytes localized to zone 2 of the hepatic acinus. Liver injury was progressive to 24 h as lipid peroxidation and hepatocellular necrosis increased. The hallmark of CCl4 hepatotoxicity, zone 3 necrosis, was observed 24 h after CCl4 administration and immune-positive hepatocytes were observed in zone 2 as well as zone 3. Immune-positive cells were no longer visible by 36 to 72 h post CCl4 administration. From 6 to 48 h after CCl4 administration, at least four adducted proteins were immune-precipitated from liver homogenates with the anti-MDA or anti-4HNE serum, which corresponded to molecular weights of 80, 150, 205, and greater than 205 kDa. These results demonstrate that 4-HNE and MDA alkylate specific hepatic proteins in a time-dependent manner, which appears to be associated with hepatocellular injury following CCl4 exposure. (C) 1999 Academic Press.