Volume 39, Issue 5, Pages 765-772 (November 2003) Mangafodipir prevents liver injury induced by acetaminophen in the mouse  Sassia Bedda, Alexis Laurent, Filomena Conti, Chistiane Chéreau, Agnès Tran, Jeanne Tran-Van Nhieu, Patrick Jaffray, Olivier Soubrane, Claire Goulvestre, Yvon Calmus, Bernard Weill, Frédéric Batteux  Journal of Hepatology  Volume 39, Issue 5, Pages 765-772 (November 2003) DOI: 10.1016/S0168-8278(03)00325-8

Fig. 1 (a) The SOD activity of Mangafodipir was evaluated using the NBT reduction technique. Results are expressed as ΔDO at 550 nm. Data are means±SEM of five independent experiments. (b) The catalase activity of Mangafodipir was determined by UV spectroscopy at 240 nm. Data are means±SEM of four independent experiments. The glutathione reductase (GR) activity of Mangafodipir was evaluated by measuring the rate of NADPH oxidation at 340 nm. Data are means±SEM of five independent experiments. (c) The spontaneous production of superoxide anions by the HUH7 hepatocytes and its inhibition by Mangafodipir and MnTBAP were evaluated using the NBT reduction technique. Data are means ΔDO at 550 nm±SEM of three independent wells. Journal of Hepatology 2003 39, 765-772DOI: (10.1016/S0168-8278(03)00325-8)

Fig. 2 Human hepatoma cell line HuH7 is protected by Mangafodipir against various oxidative stresses. Technical details are provided in Section 2. (a) Superoxide anion-oxidative stress. (b) H2O2 oxidative stress. (c) UV light stress. (d) APAP stress. Data are means±SEM of 12 independent wells in two independent experiments. Journal of Hepatology 2003 39, 765-772DOI: (10.1016/S0168-8278(03)00325-8)

Fig. 3 Kinetic analysis of GSH content in HuH7 hepatoma cells following exposure to APAP. The effects of Mangafodipir added along with APAP or 12 h later, were investigated. Means±EM of three determinations are presented. Statistical significance of differences between GSH content in the presence or in the absence of Mangafodipir, is indicated. (□) APAP+preventive Mangafodipir; (▵) APAP+curative Mangafodipir; () APAP alone. Journal of Hepatology 2003 39, 765-772DOI: (10.1016/S0168-8278(03)00325-8)

Fig. 4 (a) A single sublethal i.p. injection of 500 mg/kg (APAP500; n=6) induced significant hepatocyte cytolysis compared to animals injected with the same dose of APAP but preventively treated with Mangafodipir (n=6) or MnTBAP (n=6), whereas CuDIPS had more limited effects. (b,c) The protection conferred by Mangafodipir and MnTBAP in mice intoxicated with APAP500 was confirmed by the levels of serum LDH and ASAT. Bars represent means±SEM. Statistical evaluation was performed versus untreated animals. Journal of Hepatology 2003 39, 765-772DOI: (10.1016/S0168-8278(03)00325-8)

Fig. 5 (a) Analysis of cytochrome c release. Mitochondrial and cytosolic fractions were prepared from livers of mice intoxicated with APAP500 treated or not with CuDIPS, MnTBAP or Mangafodipir. Fractions were screened by ELISA for the detection of cytochrome c. (b) Caspase-3 activity in cytosolic fractions was measured by the release of the chromophore pNA from a peptide substrate selective for caspase-3 (DEVD-pNA). Bars represent means±SEM, six mice in each group. Statistical evaluation was performed versus untreated animals. Journal of Hepatology 2003 39, 765-772DOI: (10.1016/S0168-8278(03)00325-8)