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Overexpression and nuclear translocation of glyceraldehyde‐3‐phosphate dehydrogenase in acetaminophen acutely intoxicated rats
Author(s) -
Delfante Amalia,
Ruiz María Laura,
DiCarlo María Biatriz,
Perazzo Juan Carlos,
Mottino Aldo Domingo,
Ghanem Carolina Inés
Publication year - 2012
Publication title -
the faseb journal
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.709
H-Index - 277
eISSN - 1530-6860
pISSN - 0892-6638
DOI - 10.1096/fasebj.26.1_supplement.1050.14
Subject(s) - glyceraldehyde 3 phosphate dehydrogenase , cytosol , western blot , microbiology and biotechnology , mitochondrion , oxidative stress , glyceraldehyde , dehydrogenase , messenger rna , chemistry , acetaminophen , biochemistry , biology , enzyme , gene
The mechanism of acetaminophen (AAP) hepatotoxicity involves glutathione depletion, oxidative stress and mitochondrial dysfunction leading to necrosis. Recently, glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) was highlighted as a mediator of cell death. Aim To evaluate GAPDH expression and subcellular distribution in liver of AAP acute intoxicated rats. M&M Male Wistar rats were injected i.p. with 1 g/kg b.w. of AAP (AAP group) or vehicle (C group) (n=4). After 24 h, liver GAPDH expression (mRNA and protein) was studied by RT‐PCR and Western‐Blot (WB). Nuclear, mitochondrial, and cytosolic GAPDH content was evaluated in hepatic subcellular fractions by WB. Results were expressed as mean±SD and statistically analized with Student‐t‐test (* p<0.05 vs C). Results GAPDH mRNA (AAP:408±221*; C:100±26) and protein expression (AAP:292±127*; C:100±7) were significantly increased in treated rats. Subcellular distribution of GAPDH changed for nucleus (AAP:232±95*; C:100±49) but not for mitochondrial and cytosolic fractions. Conclusion AAP acute intoxication increased GAPDH expression due to a transcriptional mechanism, which questions its use as a housekeeping gene for either RT‐PCR or WB studies. Under AAP overdose, the GAPDH increase its content in the nucleus sugesting a role in AAP toxicity.