Nitrotyrosine-protein adducts in hepatic centrilobular areas following toxic doses of acetaminophen in mice
Nitrotyrosine-protein adducts in hepatic centrilobular areas following toxic doses of acetaminophen in mice
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DOI：
10.1021/tx9800349
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发表时间：
1998-06-01
影响因子：
4.1
通讯作者：
Mayeux, PR
中科院分区：
文献类型：
--
作者：
Hinson, JA;Pike, SL;Mayeux, PR
用毒性剂量的对乙酰氨基酚（300 mg/kg，ip）处理小鼠在4 h时显著增加了肝毒性，如肝脏小叶中心区域的组织学坏死和丙氨酸氨基转移酶（ALT）血清水平升高所证明的（从盐水处理小鼠的8 +/- 1 IU/L至对乙酰氨基酚处理小鼠的3226 +/- 892 IU/L）。硝酸盐和亚硝酸盐（一氧化氮合成的标志物）的血清水平也从盐水处理的小鼠的62 +/- 8 μ M增加到对乙酰氨基酚处理的小鼠的110 +/- 14 μ M（P < 0.05）。小鼠血清ALT水平与血清硝酸盐和亚硝酸盐水平的回归分析表明，血清ALT水平与血清硝酸盐和亚硝酸盐水平呈线性正相关，相关系数为0.9（P < 0.05）。y截距值（硝酸盐加亚硝酸盐水平）为63 +/- 15 μ M。对乙酰氨基酚中毒小鼠肝切片的免疫组织化学分析，使用抗3-硝基酪氨酸抗体表明酪氨酸硝化的小叶中心细胞的蛋白质。已证明酪氨酸硝化是通过过亚硝酸盐发生的，过亚硝酸盐是一种由一氧化氮和超氧化物极其快速反应形成的反应中间体，也是一种具有羟自由基样活性的物质。使用抗对乙酰氨基酚抗血清对肝脏切片的分析表明，小叶中心细胞也含有对乙酰氨基酚-蛋白质加合物，这是代谢产物N-乙酰基-对苯醌亚胺与蛋白质上的半胱氨酸残基的反应。这些数据与对乙酰氨基酚代谢活化导致一氧化氮和超氧化物合成增加以及过氧亚硝酸盐作为毒性的重要中间体一致。
Treatment of mice with a toxic dose of acetaminophen (300 mg/kg, ip) significantly increased hepatotoxicity at 4 h, as evidenced by histological necrosis in the centrilobular areas of the liver, and increased serum levels of alanine aminotransferase (ALT) (from 8 +/- 1 IU/L in saline-treated mice to 3226 +/- 892 IU/L in the acetaminophen-treated mice). Serum levels of nitrate plus nitrite (a marker of nitric oxide synthesis) were also increased from 62 +/- 8 mu M in saline-treated mice to 110 +/- 14 mu M in acetaminophen-treated mice (P < 0.05). Regression analysis of serum ALT levels to serum nitrate plus nitrite levels in individual mice revealed a positive, linear relationship between serum ALT levels and serum nitrate plus nitrite levels with a correlation coefficient of 0.9 (P < 0.05). The y intercept value (nitrate plus nitrite level) was 63 +/- 15 mu M. Immunohistochemical analysis of liver sections from acetaminophen-intoxicated mice using an anti-3-nitrotyrosine antibody indicated tyrosine nitration in the proteins of the centrilobular cells. Tyrosine nitration has been shown to occur by peroxynitrite, a reactive intermediate formed by an extremely rapid reaction of nitric oxide and superoxide and a species which also has hydroxyl radical-like activity. Analysis of liver sections using an anti-acetaminophen antiserum indicated the centrilobular cells also contained acetaminophen-protein adducts, a reaction of the metabolite N-acetyl-p-benzoquinone imine with cysteine residues on proteins. These data are consistent with acetaminophen metabolic activation leading to increased synthesis of nitric oxide and superoxide and to peroxynitrite as an important intermediate in the toxicity.