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J Biol Chem, Vol. 275, Issue 17, 12684-12691, April 28, 2000

Selective Involvement of Superoxide Anion, but Not Downstream Compounds Hydrogen Peroxide and Peroxynitrite, in Tumor Necrosis Factor--induced Apoptosis of Rat Mesangial Cells^*

Victoria Moreno-Manzano^§, Yoshihisa Ishikawa, Javier Lucio-Cazana^§, and Masanori Kitamura^¶

From the  Department of Medicine, University College Medical School, University College London, The Rayne Institute, London WC1E 6JJ, United Kingdom and the ^§ Department of Physiology and Pharmacology, University of Alcala de Henares, E-28871 Madrid, Spain

Tumor necrosis factor- (TNF-) induces reactive oxygen species (ROS) that serve as second messengers for intracellular signaling. Currently, precise roles of individual ROS in the actions of TNF- remain to be elucidated. In this report, we investigated the roles of superoxide anion (O₂), hydrogen peroxide (H₂O₂), and peroxynitrite (ONOO) in TNF--triggered apoptosis of mesangial cells. Mesangial cells stimulated by TNF- produced O₂ and underwent apoptosis. The apoptosis was inhibited by transfection with manganese superoxide dismutase or treatment with a pharmacological scavenger of O₂, Tiron. In contrast, although exogenous H₂O₂ induced apoptosis, TNF--triggered apoptosis was not affected either by transfection with catalase cDNA or by treatment with catalase protein or glutathione ethyl ester. Similarly, although ONOO precursor SIN-1 induced apoptosis, treatment with a scavenger of ONOO, uric acid, or an inhibitor of nitric oxide synthesis, N^G-nitro-L-argininemethyl ester hydrochloride, did not affect the TNF--triggered apoptosis. Like TNF--induced apoptosis, treatment with a O₂-releasing agent, pyrogallol, induced typical apoptosis even in the concurrent presence of scavengers for H₂O₂ and ONOO. These results suggested that, in mesangial cells, TNF- induces apoptosis through selective ROS. O₂, but not H₂O₂ or ONOO, was identified as the crucial mediator for the TNF--initiated, apoptotic pathway.

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