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Effects of moderate electrical stimulation on reactive species production by primary rat skeletal muscle cells: Cross talk between superoxide and nitric oxide production
Author(s) -
Lambertucci Rafael Herling,
dos Reis Silveira Leonardo,
Hirabara Sandro Massao,
Curi Rui,
Sweeney Gary,
PithonCuri Tania Cristina
Publication year - 2012
Publication title -
journal of cellular physiology
Language(s) - English
Resource type - Journals
SCImago Journal Rank - 1.529
H-Index - 174
eISSN - 1097-4652
pISSN - 0021-9541
DOI - 10.1002/jcp.22989
Subject(s) - superoxide , nitric oxide , xanthine oxidase , chemistry , reactive oxygen species , stimulation , biochemistry , skeletal muscle , nitrite , superoxide dismutase , xanthine , antioxidant , endocrinology , biology , enzyme , organic chemistry , nitrate
The effects of a moderate electrical stimulation on superoxide and nitric oxide production by primary cultured skeletal muscle cells were evaluated. The involvement of the main sites of these reactive species production and the relationship between superoxide and nitric oxide production were also examined. Production of superoxide was evaluated by cytochrome c reduction and dihydroethidium oxidation assays. Electrical stimulation increased superoxide production after 1 h incubation. A xanthine oxidase inhibitor caused a partial decrease of superoxide generation and a significant amount of mitochondria‐derived superoxide was also observed. Nitric oxide production was assessed by nitrite measurement and by using 4,5‐diaminofluorescein diacetate (DAF‐2‐DA) assay. Using both methods an increased production of nitric oxide was obtained after electrical stimulation, which was also able to induce an increase of iNOS content and NF‐κB activation. The participation of superoxide in nitric oxide production was investigated by incubating cells with DAF‐2‐DA in the presence or absence of electrical stimulation, a superoxide generator system (xanthine–xanthine oxidase), a mixture of NOS inhibitors and SOD‐PEG. Our data show that the induction of muscle contraction by a moderate electrical stimulation protocol led to an increased nitric oxide production that can be controlled by superoxide generation. The cross talk between these reactive species likely plays a role in exercise‐induced maintenance and adaptation by regulating muscular glucose metabolism, force of contraction, fatigue, and antioxidant systems activities. J. Cell. Physiol. 227: 2511–2518, 2012. © 2011 Wiley Periodicals, Inc.