cyclic-gmp and sodium-nitrate

Nitrate, an inorganic anion abundant in vegetables, is converted in vivo to bioactive nitrogen oxides including NO. We recently demonstrated that dietary nitrate reduces oxygen cost during physical exercise, but the mechanism remains unknown. In a double-blind crossover trial we studied the effects of a dietary intervention with inorganic nitrate on basal mitochondrial function and whole-body oxygen consumption in healthy volunteers. Skeletal muscle mitochondria harvested after nitrate supplementation displayed an improvement in oxidative phosphorylation efficiency (P/O ratio) and a decrease in state 4 respiration with and without atractyloside and respiration without adenylates. The improved mitochondrial P/O ratio correlated to the reduction in oxygen cost during exercise. Mechanistically, nitrate reduced the expression of ATP/ADP translocase, a protein involved in proton conductance. We conclude that dietary nitrate has profound effects on basal mitochondrial function. These findings may have implications for exercise physiology- and lifestyle-related disorders that involve dysfunctional mitochondria.

Topics: Adult; Cyclic GMP; Dietary Supplements; Double-Blind Method; Exercise; Humans; Ion Channels; Male; Mitochondria; Mitochondrial Proteins; Nitrates; Nitrites; Oxidative Phosphorylation; Oxygen Consumption; Uncoupling Protein 3

Nitric oxide (NO) is a critical negative regulator of platelets that is implicated in the pathology of thrombotic diseases. Platelets generate NO, but the presence and functional significance of NO synthase (NOS) in platelets is unclear. Inorganic nitrate/nitrite is increasingly being recognized as a source of bioactive NO, although its role in modulating platelets during health and vascular dysfunction is incompletely understood.. We investigated the functional significance and upstream sources of NO-cGMP signaling events in platelets by using established methods for assessing in vitro and in vivo platelet aggregation, and assessed the bioconversion of inorganic nitrate to nitrite during deficiency of endothelial NOS (eNOS).. The phosphodiesterase 5 (PDE5) inhibitor sildenafil inhibited human platelet aggregation in vitro. This inhibitory effect was abolished by a guanylyl cyclase inhibitor and NO scavengers, but unaffected by NOS inhibition. Inorganic nitrite drove cGMP-mediated inhibition of human platelet aggregation in vitro and nitrate inhibited platelet function in eNOS(-/-) mice in vivo in a model of thromboembolic radiolabeled platelet aggregation associated with an enhanced plasma nitrite concentration as compared with wild-type mice.. Platelets generate transient, endogenous cGMP signals downstream of NO that are primarily independent of NOS and may be enhanced by inhibition of PDE5. Furthermore, nitrite can generate transient NO-cGMP signals in platelets. The absence of eNOS leads to enhanced plasma nitrite levels following nitrate administration in vivo, which negatively impacts on platelet function. Our data suggest that inorganic nitrate exerts an antiplatelet effect during eNOS deficiency, and, potentially, that dietary nitrate may reduce platelet hyperactivity during endothelial dysfunction.

Topics: Animals; Blood Platelets; Cyclic GMP; Humans; Male; Mice, Inbred C57BL; Mice, Knockout; Nitrates; Nitric Oxide; Nitric Oxide Synthase Type III; Phosphodiesterase 5 Inhibitors; Platelet Aggregation; Platelet Aggregation Inhibitors; Second Messenger Systems; Sodium Nitrite; Time Factors

The effects of sodium nitroprusside (SNP) and acidified sodium nitrite (ASN) solutions, nitric oxide (NO)-donating compounds, and NG-nitro-L-arginine methyl ester (L-NAME), an inhibitor of NO synthesis, were studied on the spontaneous contractile activity of the isolated rabbit jejunum. The addition of SNP (10(-5) to 10(-3) mol/l or ASN (10(-5) to 10(-3) mol/l) the organ bath inhibited the amplitude of the spontaneous contractions in rabbit isolated jejunum in a concentration dependent fashion, while L-NAME (3 x 10(-5) to 3 x 10 mol/l) was without effect. Methylene blue (3 x 10(-7) to 3 10(-6) mol/l), which inhibits soluble guanylate cyclase, and oxyhemoglobin (10(-5) mol/l), an NO scavenger, counteracted the effects of both SNP (3 x 10(-4) mol/l) and ASN (10(-4) mol/l). The spontaneous motility of rabbit jejunum was also inhibited in a concentration-dependent manner by 8-Br-cyclic GM (10(-5) to 10(-3) mol/l), a permeable analogue of cyclic GM. These results provide evidence that exogenous NO may inhibit spontaneous contractility and that this effect might be mediated, in part, by cyclic GMP, whereas endogenous NO does not seem to play a role in the regulation of the spontaneous motility of rabbit jejunum in vitro.

Topics: Analysis of Variance; Animals; Cyclic GMP; Dose-Response Relationship, Drug; Gastrointestinal Motility; Guanylate Cyclase; In Vitro Techniques; Jejunum; Methylene Blue; Muscle Contraction; Muscle, Smooth; NG-Nitroarginine Methyl Ester; Nitrates; Nitric Oxide; Nitric Oxide Synthase; Nitroprusside; Oxyhemoglobins; Rabbits