The Effects of Palmitic Acid on Nitric Oxide Production by Rat Skeletal Muscle: Mechanism via Superoxide and iNOS Activation

Background: Increased plasma concentrations of free fatty acids (FFA) can lead to insulin resistance in skeletal muscle, impaired effects on mitochondrial function, including uncoupling of oxidative phosphorylation and decrease of endogenous antioxidant defenses. Nitric oxide (NO) is a highly diffusible gas that presents a half-life of 5-10 seconds and is involved in several physiological and pathological conditions. The effects of palmitic acid on nitric oxide (NO) production by rat skeletal muscle cells and the possible mechanism involved were investigated. Methods: Primary cultured rat skeletal muscle cells were treated with palmitic acid and NO production was assessed by nitrite measurement (Griess method) and 4,5-diaminofluorescein diacetate (DAF-2-DA) assay. Nuclear factor-kappa B (NF-kappa B) activation was evaluated by electrophoretic mobility shift assay and iNOS protein content by western blotting. Results: Palmitic acid treatment increased nitric oxide production. This effect was abolished by treatment with NOS inhibitors, L-nitro-arginine (LNA) and L-nitro-arginine methyl esther (L-NAME). NF-kappa B activation and iNOS content were increased due to palmitic acid treatment. The participation of superoxide on nitric oxide production was investigated by incubating the cells with DAF-2-DA in the presence or absence of palmitic acid, a superoxide generator system (X-XO), a mixture of NOS inhibitors and SOD-PEG (superoxide dismutase linked to polyethylene glycol). Palmitic acid and X-XO system increased NO production and this effect was abolished when cells were treated with NOS inhibitors and also with SOD-PEG. Conclusions: In summary, palmitic acid stimulates NO production in cultured skeletal muscle cells through production of superoxide, nuclear factor-kappa B activation and increase of iNOS protein content. Copyright (C) 2012 S. Karger AG, BaselFAPESPFAPESPCNPqCNPqCAPESCAPE

Control of muscular production of reactive species and cytokines by palmitic acid and eletrical stimulation: possible implications in aging.

Espécies reativas de oxigênio e nitrogênio controlam várias funções celulares, no entanto, quantidades excessivas destas espécies causam disfunções. Com o envelhecimento, há aumento da produção destas espécies, e de citocinas, que também exercem efeitos deletérios na musculatura esquelética, e da concentração plasmática de ácidos graxos. Contudo, a possível rassociação entre o aumento da produção de espécies reativas e de citocinas pelo músculo esquelético e da concentração plasmática de ácidos graxos não foi ainda investigada. O presente estudo teve como objetivo avaliar a modulação da produção muscular de espécies reativas de oxigênio e nitrogênio e de citocinas pelo ácido palmítico e pela eletroestimulação moderada. O ácido palmítico aumentou a produção de superóxido (via mitocôndria e NADPH oxidase), óxido nítrico (via iNOS) e citocinas (IL-6 e CINC-2), além de ativar o NF-kB. A eletroestimulação aumentou a produção de superóxido (via mitocôndria e sistema xantina-xantina oxidase), óxido nítrico (via iNOS) e ativou o NF-kB. Não foi observado aumento de citocinas no protocolo de eletroestimulação. O ácido palmítico, portanto, controla a produção de espécies reativas e citocinas pelo músculo. A eletroestimulação, também aumenta a produção destas espécies. Assim, o estresse oxidativo muscular é controlado pela atividade contrátil e as concentrações de ácidos graxos plasmáticos.Reactive oxygen and nitrogen species play a key role in several cell functions, however, high concentrations of these species cause cell dysfunction. High production of reactive species, and cytokines, which also induce skeletal muscle injury, and an increase of plasma concentration of fatty acids are observed in aging. However, an association between the augment in the production of reactive species and cytokines by skeletal muscle and of plasma fatty acid concentrations was not investigated yet. The aim of the present study was to evaluate the modulation of production of reactive oxygen and nitrogen species and of cytokines by palmitic acid and electrical stimulation muscle contraction. Palmitic acid increased the production of superoxide (through mitochondria and NADPH oxidase), nitric oxide (by iNOS), and citokynes (IL-6 and CINC-2), and induced NF-kB activation. The electrical stimulation-muscle contraction increased the production of superoxide (through mitochondria and xanthine-xanthine oxidase system), nitric oxide (by iNOS), and induced NF-kB activation. There was no change in the production of cytokines in the electrical stimulation protocol. In conclusion, palmitic acid raised the production of reactive oxygen and nitrogen species and cytokines by skeletal muscle. Electrical stimulation-muscle contraction also increased the production of these reactive species. Therefore, muscle oxidative stress is controlled by the muscle contractile activity and plasma fatty acid levels

Effects of moderate electrical stimulation on reactive species production by primary rat skeletal muscle cells: Cross talk between superoxide and nitric oxide production

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-?B 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 (xanthinexanthine 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: 25112518, 2012. (c) 2011 Wiley Periodicals, Inc.Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Fundacao de Amparo a Pesquisa do Estado de Sao Paulo (FAPESP)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Conselho Nacional de Desenvolvimento Cientifico e Tecnologico (CNPq)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)Coordenacao de Aperfeicoamento de Pessoal de Nivel Superior (CAPES)Canadian Institutes of Health ResearchCanadian Institutes of Health Researc

The effect of macadamia oil intake on muscular inflammation and oxidative profile kinetics after exhaustive exercise

In this study, we evaluated the onset and resolution of inflammation in control and macadamia oil-supplemented rats after a single session of exhaustive exercise. We also analysed the antioxidant enzymes catalase (CAT), glutathione peroxidase (GPx), superoxide dismutase (SOD), and thiobarbituric acid reactive substances (TBARS). Rats were supplemented for ten days with macadamia oil (25 mg/kg), which is rich in oleic and palmitoleic acids. On the 10th day, control and macadamia oil-treated rats exercised to physical exhaustion. Plasma and muscular levels of pro-inflammatory cytokines (tumour necrosis factor alpha [TNF-], interleukin-1 beta [IL-1], IL-6, cytokine-induced neutrophil chemoattractant-3 [CINC-3], macrophage inflammatory protein-3-alpha [MIP-3]) and soluble L-selectin were measured prior to, immediately after, and 2, 24, and 48 h after exercise. Our data revealed increases in the muscle concentrations of IL-1, L-selectin, CINC-3, and MIP-3 in the group supplemented with macadamia oil compared with the concentrations in the control group. CAT also increased in the treated group, which is important because pro-inflammatory cytokines precede growth factor production and might alter the muscle repair process. We concluded that inflammation resulting from exhaustive exercise was greater in animals that received macadamia oil than in control animals. Practical applications: There are potential benefits of using fatty acids in the treatment of injured muscles, and these substances have been used in several types of injured tissues. Macadamia oil intake alters muscle healing signaling by influencing the production of reactive oxygen species and inflammatory mediators. Therefore, it is reasonable to hypothesize that the intake of immunomodulatory supplements would be beneficial to the muscular recovery of an athlete following a strenuous exercise session.Fundação de Amparo a Pesquisa do Estado de São Paulo (FAPESP)Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)Cruzeiro do Sul Univ, Inst Phys Act & Sport Sci, Rua Galvao Bueno 868,13 Andar, BR-01506000 São Paulo, SP, BrazilUniv Paulista UNIP, Manaus, Amazonas, BrazilUniv Fed São Paulo UNIFESP, Dept Biociencias, São Paulo, SP, BrazilUniv Fed São Paulo UNIFESP, Dept Biociencias, São Paulo, SP, BrazilFAPESP: 14/03947-1CNPq: 307769/2014-3Web of Scienc

Fish Oil Supplementation Improves the Repeated-Bout Effect and Redox Balance in 20–30-Year-Old Men Submitted to Strength Training

Herein, we investigated the effect of fish oil supplementation combined with a strength-training protocol, for 6 weeks, on muscle damage induced by a single bout of strength exercise in untrained young men. Sixteen men were divided into two groups, supplemented or not with fish oil, and they were evaluated at the pre-training period and post-training period. We investigated changes before and 0, 24, and 48 h after a single hypertrophic exercise session. Creatine kinase (CK) and lactate dehydrogenase (LDH) activities, plasma interleukin-6 (IL-6) and C-reactive protein (CRP) levels, and the redox imbalance were increased in response to the single-bout session of hypertrophic exercises at baseline (pre-training period) and decreased during the post-training period in the control group due to the repeated-bout effect (RBE). The fish oil supplementation exacerbated this reduction and improved the redox state. In summary, our findings demonstrate that, in untrained young men submitted to a strength-training protocol, fish oil supplementation is ideal for alleviating the muscle injury, inflammation, and redox imbalance induced by a single session of intense strength exercises, highlighting this supplementation as a beneficial strategy for young men that intend to engage in strength-training programs