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Biochim Biophys Acta. 2013 Oct;1832(10):1591-604. doi: 10.1016/j.bbadis.2013.04.029. Epub 2013 May 2.

Hydrogen peroxide production regulates the mitochondrial function in insulin resistant muscle cells: effect of catalase overexpression.

Author information

1
Department of Biochemistry and Immunology, University of Sao Paulo, Ribeirão Preto, Brazil.

Abstract

The mitochondrial redox state plays a central role in the link between mitochondrial overloading and insulin resistance. However, the mechanism by which the ROS induce insulin resistance in skeletal muscle cells is not completely understood. We examined the association between mitochondrial function and H2O2 production in insulin resistant cells. Our hypothesis is that the low mitochondrial oxygen consumption leads to elevated ROS production by a mechanism associated with reduced PGC1α transcription and low content of phosphorylated CREB. The cells were transfected with either the encoded sequence for catalase overexpression or the specific siRNA for catalase inhibition. After transfection, myotubes were incubated with palmitic acid (500μM) and the insulin response, as well as mitochondrial function and fatty acid metabolism, was determined. The low mitochondrial oxygen consumption led to elevated ROS production by a mechanism associated with β-oxidation of fatty acids. Rotenone was observed to reduce the ratio of ROS production. The elevated H2O2 production markedly decreased the PGC1α transcription, an effect that was accompanied by a reduced phosphorylation of Akt and CREB. The catalase transfection prevented the reduction in the phosphorylated level of Akt and upregulated the levels of phosphorylated CREB. The mitochondrial function was elevated and H2O2 production reduced, thus increasing the insulin sensitivity. The catalase overexpression improved mitochondrial respiration protecting the cells from fatty acid-induced, insulin resistance. This effect indicates that control of hydrogen peroxide production regulates the mitochondrial respiration preventing the insulin resistance in skeletal muscle cells by a mechanism associated with CREB phosphorylation and β-oxidation of fatty acids.

KEYWORDS:

5, 5′-dithio-bis(2-nitrobenzoic acid); Akt; CAT; CREB; Catalase and muscle cells; CoQ; Cu,Zn-SOD; Cu,Zn-superoxide dismutase; DMEM; DTNB; Dulbecco's Modified Eagle's Medium; EGTA; ETF-QOR; FCCP; GAPDH; GPX; HCl; HRP; ICAT; Insulin resistance; KCl; KH(2)PO(4); MgCl(2); Mitochondria; Mn-SOD; Mn-superoxide dismutase; N-acetylcysteine; NAC; NaOH; PGC1α; PGM; PPARβ; ROS; Small interfering RNA; UCP; cAMP response element-binding; catalase; ethylene glycol tetraacetic acid; glutathione peroxidase; glyceraldehyde 3-phosphate dehydrogenase; horseradish peroxidase; hydrochloric acid; magnesium chloride; mitochondrial membrane potential (Δψ); p-trifluoromethoxyphenylhydrazone; peroxisome proliferative activated receptor-β; peroxisome proliferative activated receptor-γ (PPARγ) co-activator 1α; potassium chloride; potassium phosphate; prime growth medium (PGM); protein kinase B; reactive oxygen species; siRNA; sodium hydroxide; transfer flavoprotein quinone oxidoreductase; tricarboxylic acid cycle intermediates; ubiquinone; uncoupling protein; Δψ

PMID:
23643711
DOI:
10.1016/j.bbadis.2013.04.029
[Indexed for MEDLINE]
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