Mitochondrial superoxide production is related to the control of cytokine release from peritoneal macrophage after antioxidant treatment in septic rats

Background. Reactive oxygen species are involved in several intracellular pathways that ultimately lead to the activation of the innate immune system. In addition, oxidized proteins and lipids could stimulate cytokine release from macrophages through the activation of membrane receptors. Thus we here describe the effects of antioxidant administration to septic rats on peritoneal macrophage parameters of oxidative stress and cytokine release. Materials and methods. Peritoneal macrophages from Wistar rats subjected to cecal ligation and puncture (CLP). The animals were divided into four groups: sham operated, CLP, basic support (saline plus antibiotics), basic support plus N-acetylcysteine, and deferoxamine. Several times after CLP macrophages were cultured to the determination of thiobarbituric acid reactive species (TBARS), protein carbonyls, mitochondrial superoxide production, catalase, superoxide dismutase activities, and released cytokines. Results. Sepsis increased TBARS, protein carbonyls, and mitochondrial superoxide production in macrophages and this was associated with an increase release of pro-inflammatory cytokines. Basic support reversed TBARS and protein carbonyls content, but not mitochondrial superoxide production. The addition of antioxidants prevented all oxidative parameters in macrophages, and this was associated with lower cytokine release. Catalase and superoxide dismutase were modulated in the basic support group, but not in the antioxidant treated animals. Conclusions. Mitochondrial superoxide production seemed to be the differential oxidative parameter associated with antioxidant-induced modulation of cytokine release. (c) 2007 Elsevier Inc. All rights reserved.