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Type: Artigo de periódico
Title: Formation Of Hydrogen Peroxide And Nitric Oxide In Rat Skeletal Muscle Cells During Contractions.
Author: Silveira, Leonardo R
Pereira-Da-Silva, Lucia
Juel, Carsten
Hellsten, Ylva
Abstract: We examined intra- and extracellular H(2)O(2) and NO formation during contractions in primary rat skeletal muscle cell culture. The fluorescent probes DCFH-DA/DCFH (2,7-dichlorofluorescein-diacetate/2,7-dichlorofluorescein) and DAF-2-DA/DAF-2 (4,5-diaminofluorescein-diacetate/4,5-diaminofluorescein) were used to detect H(2)O(2) and NO, respectively. Intense electrical stimulation of muscle cells increased the intra- and extracellular DCF fluorescence by 171% and 105%, respectively, compared with control nonstimulated cells (p <.05). The addition of glutathione (GSH) or Tiron prior to electrical stimulation inhibited the intracellular DCFH oxidation (p <.05), whereas the addition of GSH-PX + GSH inhibited the extracellular DCFH oxidation (p <.05). Intense electrical stimulation also increased (p <.05) the intra- and extracellular DAF-2 fluorescence signal by 56% and 20%, respectively. The addition of N(G)-nitro-L-arginine (L-NA) completely removed the intra- and extracellular DAF-2 fluorescent signal. Our results show that H(2)O(2) and NO are formed in skeletal muscle cells during contractions and suggest that a rapid release of H(2)O(2) and NO may constitute an important defense mechanism against the formation of intracellular (*)OH and (*)ONOO. Furthermore, our data show that DCFH and DAF-2 are suitable probes for the detection of ROS and NO both intra- and extracellularly in skeletal muscle cell cultures.
Subject: 1,2-dihydroxybenzene-3,5-disulfonic Acid Disodium Salt
Cells, Cultured
Electric Stimulation
Enzyme Inhibitors
Fluorescent Dyes
Hydrogen Peroxide
Hydroxyl Radical
Indicators And Reagents
Muscle Contraction
Muscle, Skeletal
Nitric Oxide
Oxidative Stress
Peroxynitrous Acid
Rats, Wistar
Citation: Free Radical Biology & Medicine. v. 35, n. 5, p. 455-64, 2003-Sep.
Rights: fechado
Date Issue: 2003
Appears in Collections:Unicamp - Artigos e Outros Documentos

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