Measurement of peroxiredoxin-4 serum levels in rat tissue and its use             as a potential marker for hepatic disease.
Peroxiredoxin (Prx)-4, a secretable endoplasmic reticulum (ER)-resident isoform of the mammalian Prx family, functions as a thioredoxin-dependent peroxidase. It is acknowledged that Prx-4 plays a role in the detoxification of hydrogen peroxide, and potentially other peroxides, which may be generated during the oxidative folding of proteins and oxidative stress in the ER. The present study was undertaken in order to specifically quantify the tissue levels of Prx-4. To accomplish this, an enzyme-linked immunosorbent assay was developed using a specific polyclonal antibody produced by immunizing a rabbit with native recombinant rat Prx-4 protein. The assay was used to detect Prx-4 in the range of 0.1 and 10 ng/ml, and to investigate tissue distribution in rats. Using this immunoassay, we found that the serum levels of Prx-4 were substantially lower in asymptomatic Long-Evans Cinnamon rats, a rat model of Wilson's disease, compared to normal rats. In addition, the treatment of rat hepatoma cells with N-acetylcysteine led to a significant increase in the release of Prx-4 protein into the medium; thus, it appears likely that the secretion of Prx-4 is associated with the redox state within cells. These results suggest that serum Prx-4 has potential for use as a biomarker for hepatic oxidative stress.
Division of Molecular Cell Biology, Department of Biomolecular                     Sciences, Faculty of Medicine, Saga University, Saga 849‑8501, Japan.
Ritsu
R
Ito
Motoko
M
Takahashi
Hideyuki
H
Ihara
Hiroki
H
Tsukamoto
Junichi
J
Fujii
Yoshitaka
Y
Ikeda
D000111:Acetylcysteine / Q000494:pharmacology; D000818:Animals; D000906:Antibodies / Q000276:immunology / Q000378:metabolism; D000918:Antibody Specificity; D014408:Biomarkers, Tumor / Q000097:blood; D006528:Carcinoma, Hepatocellular / Q000378:metabolism / Q000473:pathology; D045744:Cell Line, Tumor; D003470:Culture Media / Q000378:metabolism; D003921:Diabetes Mellitus, Experimental / Q000378:metabolism / Q000473:pathology; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods; D005260:Female; D007074:Immunoglobulin G / Q000097:blood; D008099:Liver / Q000378:metabolism / Q000473:pathology; D008107:Liver Diseases / Q000175:diagnosis* / Q000378:metabolism / Q000473:pathology; D008297:Male; D010084:Oxidation-Reduction; D018384:Oxidative Stress; D054464:Peroxiredoxins / Q000097:blood* / Q000276:immunology / Q000378:metabolism; D011817:Rabbits; D051381:Rats; D020332:Rats, Inbred LEC; D017207:Rats, Sprague-Dawley; D017208:Rats, Wistar; D011994:Recombinant Proteins / Q000008:administration & dosage / Q000276:immunology / Q000378:metabolism; D014611:Vaccination
D000906:Antibodies; D014408:Biomarkers, Tumor; D003470:Culture Media; D007074:Immunoglobulin G; D011994:Recombinant Proteins; D054464:Peroxiredoxins; C515982:Prdx4 protein, rat; D000111:Acetylcysteine