Use of proteomic methods to identify serum biomarkers associated with rat liver toxicity or hypertrophy
Use of proteomic methods to identify serum biomarkers associated with rat liver toxicity or hypertrophy
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DOI：
10.1373/clinchem.2005.049908
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发表时间：
2005-10-01
影响因子：
9.3
通讯作者：
Townsend, RR
中科院分区：
文献类型：
--
作者：
Amacher, DE;Adler, R;Townsend, RR
背景：我们的目标是在大鼠中识别可能作为肝肿大、肝细胞坏死或肝胆损伤的敏感指标的血清标记蛋白，并使用它们来分析来自合作蛋白质组学项目的数据。方法：在组成合作项目的4项研究中，每项研究都给大鼠提供4种通过不同机制靶向肝脏的化合物中的1种。在给药后3个时间点中的1个时间点采集血清和肝脏样本。用双向凝胶电泳法(2DGE)去除血清中的主要分泌蛋白，将其分离为蛋白质组分。同时对肝脏标本进行2DGE处理。对药物治疗后数量显著增加或减少的蛋白质斑点进行回收、消化、质谱分析，并与现有数据库进行比较以进行鉴定。进一步考虑的标准是(A)暂时性表达(即早期、暴发期或恢复期的增加或减少)，(B)已知的生物学功能，(C)可能的肝脏起源，以及(D)在已发表的研究中任何先前与毒性有关的因素。结果：维生素D结合蛋白、对氧磷酶、细胞视黄醇结合蛋白、苹果酸脱氢酶、F蛋白和嘌呤核苷磷酸化酶被确定为药物治疗大鼠肝脏效应的经验性血清标志物。结论：蛋白质组学可用于毒物暴露后肝功能改变的外周生物标志物的鉴定和确认。(C)2005年美国临床化学协会。
Background: Our objectives were to identify serum marker proteins in rats that might serve as sensitive indicators of hepatomegaly, hepatocellular necrosis, or hepatobiliary injury and to use them to analyze data from a collaborative proteomics project.Methods: In each of 4 studies comprising the collaborative project, rats were given 1 of 4 compounds that target the liver through different mechanisms. Sera and liver samples were collected by terminal bleeds at 1 of 3 postdose time points. Sera were depleted of major secretory proteins and then separated into,protein features by 2-dimensional gel electrophoresis (2DGE). Liver specimens were also processed and subjected to 2DGE. Protein spots that significantly increased or decreased in quantity after drug treatment were recovered, digested, analyzed by mass spectroscopy, and compared with available databases for identification. Criteria for further consideration were (a) temporal expression (i.e., increase or decrease at early, fulminant, or recovery periods), (b) known biological function, (c) probable hepatic origin, and (d) any previous association with toxicity in published studies. Markers that changed significantly at the early time point were important because of their potential sensitivity for signaling minimal damage.Results: Vitamin D-binding protein, paraoxonase, cellular retinol-binding protein, malate dehydrogenase, F-protein, and purine nucleoside phosphorylase were identified as empirically confirmed serum markers for hepatic effects in drug-treated rats.Conclusion: Proteomics can be applied for the identification and confirmation of peripheral biomarkers for altered liver function after toxicant exposure. (c) 2005 American Association for Clinical Chemistry.