Glutathione redox status in hippocampi of mice undergoing 2- or 4-mo moderate treadmill running.

<p>Evaluation of the glutathione redox balance in hippocampi of mature CD1 female mice undergoing a two- or four-month moderate and regular treadmill-based exercise program (E2 or E4, respectively); age-matched sedentary animals (S2, S4) were used as controls (n = 12 per group). A significant age-dependent decrease in the total vs oxidized glutathione ratio was found within mouse hippocampal formations (S4 vs S2). 2-mo running decreased the total vs disulfide glutathione ratio (E2 vs S2), whereas no significant change was induced by 4-mo exercise (E4 vs S4). Values were given as means ± std. dev. The level of statistical significance was computed by using two-way ANOVA and post-hoc Newman-Keuls test: *** P<0.001; ** P<0.01. Experiments were performed in triplicate.</p

NAD redox balance and expression of SIRT1 in hippocampi of mice undergoing 2- or 4-mo moderate treadmill running.

<p>Assessment of nicotinamide adenine dinucleotide redox balance (panel a) and Western immunoblot against SIRT1 (panel b) in hippocampi of mature CD-1 female mice undergoing a two- or four-month moderate and regular treadmill-based exercise program (E2 or E4, respectively); age-matched sedentary animals (S2, S4) were used as controls (n = 12 per group). Both the NAD redox ratio and the SIRT1 protein level were decreased in an age-dependent fashion (S4 vs S2) (panels a and b, respectively). 2-mo exercise reduced both the NAD⁺/NADH ratio and the expression level of SIRT1 (E2 vs S2) (panels a and b, respectively), whereas both the NAD redox balance and the protein level of SIRT1 increased after 4-mo exercise (E4 vs S4) (panels a and b, respectively). Western blot signals were normalized against the housekeeping β-actin. Representative western immunoblots of three independent experiments were reported in panel b. Values were given as means ± std. dev. The level of statistical significance was computed by using two-way ANOVA and post-hoc Newman-Keuls test: *** P<0.001; ** P<0.01. Experiments were performed in triplicate.</p

MG-related protein damage and expression of MG-targeting detoxification system in hippocampi of mice undergoing 2- or 4-mo moderate treadmill running.

<p>Immunoreactivity levels against arg-pyrimidine and protein expression of glyoxalase 1 (GLO1) in hippocampi of mature CD-1 female mice undergoing a two- or four-month moderate and regular treadmill-based exercise program (E2 or E4, respectively); age-matched sedentary animals (S2, S4) were used as controls (n = 12 per group). As shown in panel a, an age-dependent increase in the levels of MG-damaged protein was detected (S4 vs S2), and an increasing trend in MG-related protein damage was observed after 2 months of running (E2 vs S2). The immunoreactivity against arg-pyrimidine decreased after 4 months of regular exercise (E4 vs S4). Dot-blots signals were normalized against the Comassie Brilliant Blue-based total protein staining. In the panel a, right section, representative dot immunoblots of three independent experiments were reported. As reported in panel b, an age-dependent increase in the GLO1 protein expression levels was detected (S4 vs S2). 2-mo exercise increased GLO1 protein levels (E2 vs S2), while the expression levels of GLO1 decreased after four months of regular running (E4 vs S4). Immunosignals were normalized against the housekeeping β-actin. Representative western immunoblots of three independent experiments were reported in panel b. Values were given as means ± std. dev. The level of statistical significance was computed by using two-way ANOVA and post-hoc Newman-Keuls test: *** P<0.001; ** P<0.01. Experiments were performed in triplicate.</p

Hippocampal peroxidative damage in mice undergoing 2- or 4-mo moderate treadmill running.

<p>Assessment of hippocampal levels of thiobarbituric acid-reactive substances (TBARS) levels in mature CD-1 female mice undergoing a two- or four-month moderate and regular treadmill-based exercise program (E2 or E4, respectively); age-matched sedentary animals (S2, S4) were used as controls (n = 12 per group). TBARS levels were found to be increased by the sedentary aging (S4 vs S2). 2-mo exercise increased TBARS hippocampal concentrations (E2 vs S2), while 4-mo running decreased TBARS levels (E4 vs S4). Values were given as means ± std. dev. The level of statistical significance was computed by using two-way ANOVA and post-hoc Newman-Keuls test: *** P<0.001; Experiments were performed in triplicate.</p

Ponderal profiles of mice undergoing a long-term moderate treadmill running.

<p>No significant age-dependent main effect was detected on body weight, neither did treadmill running induce significant alterations in ponderal state. Values were given as means ± std. dev.</p

Brain molecular damage of mice undergoing a long-term moderate treadmill running.

<p>Thiobarbituric acid-reactive substances (TBARS, panel a) levels and protein carbonyl content (PCC, panel b) in cortices of mice undergoing a two- or four-month moderate and regular treadmill-based exercise program (E2 or E4, respectively); age-matched sedentary animals (S2, S4) were used as controls (n = 6 per group). No significant age-dependent change was observed in cortical TBARS concentrations (S4 vs S2), while two months of exercise elevated significantly TBARS amounts (E2 vs S2). Four-month exercised and unexercised mice showed similar TBARS levels (E4 vs S4). The PCC increased in an age-dependent manner (S4 vs S2). Two months of exercise elevated PCC, in comparison to sedentary animals (E2 vs S2). Conversely, PCC content was significantly reduced in four-month-exercised mice, when compared to age-matched unexercised animals (E4 vs S4). Values were given as means ± std. dev. The level of statistical significance was computed by using two-way ANOVA and post-hoc Newman-Keuls test: *** P<0.001; ** P<0.01. Experiments were performed in triplicate.</p

Neurotrophic support in brains of mice undergoing a long-term moderate treadmill running.

<p>Western immunoblot against the brain derived neurotrophic factor (BDNF) (panel a) and the phosphorilated cAMP response element binding (p-CREB) transcriptional activator (panel b) in brain cortices of adult CD1 female mice undergoing a two- or four-month moderate and regular treadmill-based exercise program (E2 or E4, respectively); age-matched sedentary animals (S2, S4) were used as controls (n = 6 per group). A significant age-dependent variation in BDNF protein expression was detected (S4 vs S2). BDNF levels were also significantly reduced in two-month exercised mice (E2 vs S2), yet strongly enhanced in four-month exercised mice (E4 vs S4), with respect to age-matched counterparts. Significant age-dependent reductions in the protein levels of p-CREB (S4 vs S2). Two months of exercise caused a conspicuous decrease in p-CREB protein levels (E2 vs S2), whereas p-CREB protein amounts increased after four months of regular running (E4 vs S4), with respect to age-matched unexercised mice. Immunosignals were normalized against the housekeeping β-actin. Representative PVDF-western immunoblots of three independent experiments are reported. Values were given as means ± std. dev. The level of statistical significance was computed by using two-way ANOVA and post-hoc Newman-Keuls test: * P<0.05; ** P<0.01; *** P<0.001. Experiments were performed in triplicate.</p

Methylglyoxal-related enzymatic removal and molecular damage in brains of mice undergoing a long-term moderate treadmill running.

<p>Immunoreactivity levels against arg-pyrimidine (arg-pyr, panel a) and specific activities of glyoxalase 1 (GLO1, panel b) and glyoxalase 2 (GLO2, panel c) in brain cortices of adult CD1 female mice undergoing a two- or four-month moderate and regular treadmill-based exercise program (E2 or E4, respectively); age-matched sedentary animals (S2, S4) were used as controls (n = 6 per group). Dot blot assays revelead an age-dependent increase in the levels of MG-damaged proteins (S4 vs S2). No significant variation of MG-damaged polypeptides was detected after two months of exercise (E2 vs S2), while immunoreactivity against MG-modified arginine residues decreased markedly after four-month physical activity, with respect to age-matched unexercised mice (E4 vs S4). Immunosignals were normalized against the Coomassie Brilliant Blue (CBB)-based total protein staining. In the panel a, right section, representative PVDF-dot immunoblots of three independent experiments are reported, together with analyses for total protein loading performed by the ImageJ software. Both GLO1 and GLO2 specific activities increased in an age-dependent manner (S4 vs S2). The exercise program elevated GLO2 activity exclusively after two months of regular exercise (E2 vs S2), whereas GLO1 specific activity was increased only after four months of physical activity (E4 vs S4). Values were given as means ± std. dev. The level of statistical significance was computed by using two-way ANOVA and post-hoc Newman-Keuls test: * P<0.05; *** P<0.001. Experiments were performed in triplicate.</p

Neurotrophic support in brains of mice undergoing a long-term moderate treadmill running.

<p>Western immunoblot against the activated high affinity tyrosinekinase B receptor (p-TrkB) in brain cortices of adult CD1 female mice undergoing a two- or four-month moderate and regular treadmill-based exercise program (E2 or E4, respectively); age-matched sedentary animals (S2, S4) were used as controls (n = 6 per group). Significant age-dependent reductions in the protein amounts of p-TrkB (S4 vs S2) were detected. Two months of exercise caused a conspicuous decrease in the expression levels of p-TrkB, with respect to age-matched unexercised mice (E2 vs S2), whereas protein levels of the activated receptor were significantly elevated after four-month physical activity, when compared to sedentary animals (E4 vs S4). Immunosignals were normalized against the housekeeping β-actin. In the lower sections, representative PVDF-western immunoblots of three independent experiments are reported. Values were given as means ± std. dev. The level of statistical significance was computed by using two-way ANOVA and post-hoc Newman-Keuls test: ** P<0.01; *** P<0.001. Experiments were performed in triplicate.</p

Antioxidant enzymatic defense in brains of mice undergoing a long-term moderate treadmill running.

<p>Specific activities of superoxide dismutase (SOD) (panel a), catalase (CAT) (panel b) and glutathione peroxidase (GPX) (panel c) in brain cortices of adult CD1 female mice undergoing a two- or four-month moderate and regular treadmill-based exercise program (E2 or E4, respectively); age-matched sedentary animals (S2, S4) were used as controls (n = 6 per group). No major age-dependent variation of tSOD and CAT specific activities was revealed; however, a significant increase in GPX activity was detected when comparing S4 and S2. Two-month physical activity reduced specific activities of tSOD, CAT and GPX catalytic capacities (E2 vs S2), whereas four-month exercise triggered a significant elevation of tSOD and CAT (E4 vs S4). Values were given as means ± std. dev. The level of statistical significance was computed by using two-way ANOVA and post-hoc Newman-Keuls test: * P<0.05; ** P<0.01; *** P<0.001. Experiments were performed in triplicate.</p