Abstract
Changes in the serum proteome were identified during early, fulminant, and recovery phases of liver injury from acetaminophen in the rat. Male F344 rats received a single, noninjury dose or a high, injury-producing dose of acetaminophen for evaluation at 6 to 120 h. Two-dimensional gel electrophoresis of immunodepleted serum separated approximately 800 stained proteins per sample from which differentially expressed proteins were identified by mass spectrometry. Serum alanine aminotransferase/aspartate aminotransferase levels and histopathology revealed the greatest liver damage at 24 and 48 h after high-dose acetaminophen corresponding to the time of greatest serum protein alterations. After 24 h, 68 serum proteins were significantly altered of which 23 proteins were increased by >5-fold and 20 proteins were newly present compared with controls. Only minimal changes in serum proteins were noted at the low dose without any histopathology. Of the 54 total protein isoforms identified by mass spectrometry, gene ontology processes for 38 unique serum proteins revealed involvement of acute phase response, coagulation, protein degradation, intermediary metabolism, and various carrier proteins. Elevated serum tumor necrosis factor-α from 24 to 48 h suggested a mild inflammatory response accompanied by increased antioxidant capability demonstrated by increased serum catalase activity. Antibody array and enzyme-linked immunosorbent assay analyses also showed elevation in the chemokine monocyte chemoattractant protein-1 and the metalloprotease inhibitor tissue inhibitor of metalloproteinases-1 during this same period of liver injury. This study demonstrates that serum proteome alterations probably reflect both liver damage and a concerted, complex response of the body for organ repair and recovery during acute hepatic injury.
Footnotes
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This work was supported by the Intramural Research program of the National Institutes of Health, National Institute of Environmental Health Sciences. This project was also funded in part with federal funds from the National Institute of Environmental Health Sciences, National Institutes of Health, under Contract N01-ES-25495.
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Article, publication date, and citation information can be found at http://jpet.aspetjournals.org.
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doi:10.1124/jpet.106.102681.
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ABBREVIATIONS: TNF-α, tumor necrosis factor-α; LC-MS/MS, liquid chromatography-tandem mass spectrometry; CMC, carboxymethylcellulose; ELISA, enzyme-linked immunosorbent assay; MCP-1, monocyte chemoattractant protein-1; IPG, immobilized pH gradient; ALT, alanine aminotransferase; AST, aspartate aminotransferase; PAGE, polyacrylamide gel electrophoresis; CHAPS, 3-[(3-cholamidopropyl)-dimethylammonio]-1-propanesulphonate; MALDI, matrix-assisted laser desorption ionization; ANOVA, analysis of variance; ARP-3, actin-related protein-3; GSH, glutathione; TIMP-1, tissue inhibitor of metalloproteinases-1; LPS, lipopolysaccharide; HGF, hepatic growth factor; APAP, acetaminophen.
- Received February 9, 2006.
- Accepted May 8, 2006.
- The American Society for Pharmacology and Experimental Therapeutics
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