Acetaminophen (APAP) hepatocyte toxicity is associated with increased translocation of glyceraldehyde-3-phosphate dehydrogenase (GAPDH) to the nucleus.

CIRIACI N; VIGO MB; RIGALLI JP; MOTTINO AD; RUIZ ML; GHANEM CI

Boston

Congreso; Experimental Biology 2015; 2015

American society of Pharmaceutical and Experimental Therapeutic

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 IC50 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