Acute exercise induce endothelial nitric oxide synthase phosphorylation via Akt and AMP-activated protein kinase in aorta of rats: Role of reactive oxygen.

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Acute exercise induce endothelial nitric oxide synthase phosphorylation via Akt and AMP-activated protein kinase in aorta of rats: Role of reactive oxygen species Viviane A. Barbosa, Thais F. Luciano, Scherolin O. Marques, Marcelo F. Vitto, Daniela R. Souza, Luciano A. Silva, João Paulo A. Santos, José C. Moreira, Felipe Dal-Pizzol, Fabio S. Lira, Ricardo A. Pinho, Claudio T. De Souza International Journal of Cardiology Volume 167, Issue 6, Pages 2983-2988 (September 2013) DOI: 10.1016/j.ijcard.2012.08.050 Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions

Fig. 1 Effects ex vivo of H2O2 on Akt, AMPK, eNOS and VASP phosphorylation in freshly isolated aorta. The aortic tissue was pre-incubated for 30min without (control and H2O2 groups) and with: N-acetylcysteine, LY294002, Compound C or LY294002 plus Compound C. After this period, the aorta of all groups (except control group) were incubated with 200μM H2O2 for 30min. Immediately after incubation, aortas were homogenized and immunoblot performed to measure the levels of the phosphorylation (top panel) of Akt (A), AMPK (B), eNOS (C) and VASP (D). The lower panel shows bands representative of Akt, AMPK, eNOS and VASP protein levels. The results are presented as arbitrary units. Bars are presented as means of area of bands±SEM of five different incubation experiments per group, *p<0.05, H2O2 group versus control and #p<0.05, NAC and inhibitors treatments versus H2O2 group. International Journal of Cardiology 2013 167, 2983-2988DOI: (10.1016/j.ijcard.2012.08.050) Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions

Fig. 2 Superoxide levels and DCF concentration in aorta of rats. Superoxide levels (A) and diclorofluorescein concentration (B). Superoxide levels are expressed as nmol/min/mg of protein (A) and DCF is expressed as μmol/mg of protein (B). Bars represent means±SEM of five rats per group. *p<0.05, exercise versus control group; #p<0.05, exer+NAC versus exercise group. International Journal of Cardiology 2013 167, 2983-2988DOI: (10.1016/j.ijcard.2012.08.050) Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions

Fig. 3 Effect in vivo of acute exercise on Akt, AMPK, eNOS and VASP phosphorylation in aorta of rats. The levels of the phosphorylation (panel upper) the Akt (A), AMPK (B), eNOS (C) and VASP (D) were measured by immunoblot in aorta of rats killed immediately after acute exercise. The lower panel shows bands representative of Akt, AMPK and eNOS protein levels. The results are presented as arbitrary units. Bars are presented as means of area of bands±SEM of five rats per group, *p<0.05, exercise versus control group; #p<0.05, exer+NAC versus exercise group. International Journal of Cardiology 2013 167, 2983-2988DOI: (10.1016/j.ijcard.2012.08.050) Copyright © 2012 Elsevier Ireland Ltd Terms and Conditions