N-Acetylcysteine Partially Reverses Oxidative Stress and Apoptosis Exacerbated by Mg-Deficiency Culturing Conditions in Primary Cultures of Rat and Human Hepatocytes

N-Acetylcysteine Partially Reverses Oxidative Stress and Apoptosis Exacerbated by Mg-Deficiency Culturing Conditions in Primary Cultures of Rat and Human Hepatocytes

Hélène Martin, PhD, Catherine Abadie, PhD, Bruno Heyd, MD, PhD, Georges Mantion, MD, PhD, Lysiane Richert, PhD and Alain Berthelot, PhD

Laboratoire de Biologie Cellulaire (H.M., L.R.)
Laboratoire de Physiologie (A.B.)
Kaly-Cell (C.A.), UFR des Sciences Médicales et Pharmaceutiques, Service de Chirurgie Digestive et Vasculaire
Hôpital Jean Minjoz (B.H., G.M.), Besançon, FRANCE

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Fig. 1. Effects of extracellular magnesium on caspase-3 activation. (A) Time course analysis in a primary culture of human hepatocytes (HH1) exposed to 0, 0.2, 0.4 and 0.8 mM magnesium. (B) Concentration-dependent analysis in three representative primary cultures of human hepatocytes (HH1, HH2 and HH5) and cultures of rat hepatocytes (RH, expressed as means + SEM, n = 3 different hepatocyte cultures, means without a common letter (a, b, c) differ, p $≤$ 0.05) exposed to 0, 0.2, 0.4 and 0.8 mM magnesium for 72 h. Caspase-3 activities in the 0.8 mM condition are 191.8, 90.9, 243.1 and 9.1 pmol AMC/min/mg protein for HH1, HH2, HH5 and RH, respectively.

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Fig. 2. Effects of extracellular magnesium on GSH concentration. Concentration-dependent analysis in three primary cultures of human hepatocytes (HH2, HH3 and HH4) and cultures of rat hepatocytes (RH, expressed as means + SEM, n = 3 different hepatocyte cultures, means without a common letter (a, b, c) differ p $≤$ 0.05) exposed to 0, 0.2, 0.4 and 0.8 mM magnesium for 72 h. GSH concentrations in the 0.8 mM condition are 48.7, 66.5, 30.4 and 23.8 nmol/mg protein for HH2, HH3, HH4 and RH, respectively.

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Fig. 3. Effect of NAC supplementation in Mg deficient and Mg standard cultures of rat hepatocytes, on reduced glutathione (GSH), caspase-3 activity and thiobarbituric acid reactive substances (TBARS) concentrations. ${blacksquare}$ control cultures and ${square}$ cultures treated with NAC (5 mM). Values are means ± S.E.M. (n = 3 different hepatocyte cultures). For control cultures, means without a common letter (a, b, c) differ (p $≤$ 0.05). For NAC cultures, means without a common letter (a', b', c') differ (p $≤$ 0.05). *Different from control cultures without NAC in the corresponding Mg treatment condition (p $≤$ 0.05).