PT  - JOURNAL ARTICLE
AU  - Xi Yang
AU  - James Greenhaw
AU  - Qiang Shi
AU  - Dean W. Roberts
AU  - Jack A. Hinson
AU  - Levan Muskhelishvili
AU  - Kelly Davis
AU  - William F. Salminen
TI  - Mouse Liver Protein Sulfhydryl Depletion after Acetaminophen Exposure
AID  - 10.1124/jpet.112.199067
DP  - 2013 Jan 01
TA  - Journal of Pharmacology and Experimental Therapeutics
PG  - 286--294
VI  - 344
IP  - 1
4099  - http://jpet.aspetjournals.org/content/344/1/286.short
4100  - http://jpet.aspetjournals.org/content/344/1/286.full
SO  - J Pharmacol Exp Ther2013 Jan 01; 344
AB  - Acetaminophen (APAP)-induced liver injury is the leading cause of acute liver failure in many countries. This study determined the extent of liver protein sulfhydryl depletion not only in whole liver homogenate but also in the zonal pattern of sulfhydryl depletion within the liver lobule. A single oral gavage dose of 150 or 300 mg/kg APAP in B6C3F1 mice produced increased serum alanine aminotransferase levels, liver necrosis, and glutathione depletion in a dose-dependent manner. Free protein sulfhydryls were measured in liver protein homogenates by labeling with maleimide linked to a near infrared fluorescent dye followed by SDS-polyacrylamide gel electrophoresis. Global protein sulfhydryl levels were decreased significantly (48.4%) starting at 1 hour after the APAP dose and maintained at this reduced level through 24 hours. To visualize the specific hepatocytes that had reduced protein sulfhydryl levels, frozen liver sections were labeled with maleimide linked to horseradish peroxidase. The centrilobular areas exhibited dramatic decreases in free protein sulfhydryls while the periportal regions were essentially spared. These protein sulfhydryl-depleted regions correlated with areas exhibiting histopathologic injury and APAP binding to protein. The majority of protein sulfhydryl depletion was due to reversible oxidation since the global- and lobule-specific effects were essentially reversed when the samples were reduced with tris(2-carboxyethy)phosphine before maleimide labeling. These temporal and zonal pattern changes in protein sulfhydryl oxidation shed new light on the importance that changes in protein redox status might play in the pathogenesis of APAP hepatotoxicity.