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Acetaminophen (APAP) hepatocyte toxicity is associated with increased translocation of glyceraldehyde‐3‐phosphate dehydrogenase (GAPDH) to the nucleus.
Author(s) -
Ciriaci Nadia,
Vigo Belén,
Rigalli Juan,
Mottino Aldo,
Ruiz María,
Ghanem Carolina
Publication year - 2015
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.29.1_supplement.937.8
Subject(s) - glyceraldehyde 3 phosphate dehydrogenase , cytosol , viability assay , hepatocyte , apoptosis , acetaminophen , chemistry , chromosomal translocation , cytochrome c , microbiology and biotechnology , pharmacology , in vitro , biology , dehydrogenase , biochemistry , enzyme , gene
Previously, we have described that APAP intoxication induces nuclear translocation of GAPDH in rat liver. Recently, GAPDH nuclear traslocation was highlighted as a mediator of cell death. Aim To test if GAPDH nuclear translocation is associated with loss of viability of rat hepatocytes exposed to APAP. M&M: Rat isolated hepatocytes were exposed to different doses of APAP (0‐50 mM), for 24 h, in the absence or presence of RAS (10 µM), an inhibitor of GAPDH nuclear translocation. Cell viability was evaluated by the MTT assay and IC 50 was calculated. Nuclear and cytosolic GAPDH, cytosolic cytochrome c, and mitochondrial BAX contents were evaluated by WB in hepatocytes treated with APAP (30 mM) and/or RAS (10 µM) for 24 h. Results IC50: RAS: 40.0±8.5 mM*; Control: 18.5±0.6 mM. Nuclear/Nuclear+cytosolic GAPDH expression ratio: APAP: 155±29* $ ; APAP‐RAS: 26±10*; RAS: 42±17*; C: 100±5. Cytosolic cytochrome c: APAP: 320±7.5* $ ; APAP‐RAS: 31±22; RAS: 19±32; C: 100±59. Mitochondrial BAX: APAP: 468±57* ♦ ; APAP‐RAS: 342±45* ♦ ; RAS: 152±81; C: 100±59. WB data are expressed as % of control. * P<0.05 vs C; $ P<0.05 vs RAS and APAP‐RAS. ♦ P<0.05 vs RAS. Conclusion APAP acute intoxication increased GAPDH nuclear translocation in isolated hepatocytes. RAS blockage of such traslocation was associated with prevention in alteration of apoptotic/necroptotic marker (cytochrome C) and consequently with increased survival of the cells to APAP cytotoxicity. The data strongly suggest a link between APAP liver toxicity and GAPDH translocation.