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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 L.R.
Pereira-Da-Silva L.
Juel C.
Hellsten Y.
Abstract: We examined intra- and extracellular H 2O 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 2O 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 2O 2 and NO are formed in skeletal muscle cells during contractions and suggest that a rapid release of H 2O 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. © 2003 Elsevier Inc.
Rights: fechado
Identifier DOI: 10.1016/S0891-5849(03)00271-5
Date Issue: 2003
Appears in Collections:Unicamp - Artigos e Outros Documentos

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