RESUMEN
The new glucosyl sarpagan alkaloid designated as 21(R*)-(O-ß-glucosyl)-hydroxy-sarpagan-17-oic acid, along with eleven known alkaloids were isolated from a soluble alkaloidal fraction from the ethanol extract of Rauvolfia ligustrina. Their structures were elucidated by interpretation of spectroscopic data (1D and 2D NMR), HRESIMS experiment, GIAO 13C NMR calculations, and comparison with literature data. All the isolated alkaloids were screened by their neuroinhibitory effects using the electrically stimulated mice vas deferens bioassay. Compounds 1, 2 and 9 presented a potent inhibitory effect in the neurotransmission while 3 and 11 showed an acute neuroexcitatory effect. Compound 10 exhibited a very effective post-synaptic inhibitory activity.
Asunto(s)
Alcaloides Indólicos/farmacología , Raíces de Plantas/química , Rauwolfia/química , Transmisión Sináptica/efectos de los fármacos , Animales , Brasil , Estimulación Eléctrica , Técnicas In Vitro , Alcaloides Indólicos/química , Espectroscopía de Resonancia Magnética , Masculino , Ratones , Estructura Molecular , Fitoquímicos/aislamiento & purificación , Fitoquímicos/farmacología , Extractos Vegetales/química , Conducto Deferente/efectos de los fármacosRESUMEN
We examined whether off-season (OffS) and pre-contest (PreC) periods affect blood oxidative stress, inflammatory, immunological, and psychological markers in 20 bodybuilders. The athletes recorded their food intake (3-day record), physical activities, mood states (Profile of Mood States, POMS), recovery-stress (Recovery-Stress Questionnaire for Athletes, RESTQ-Sport), and upper respiratory symptoms (Wisconsin Upper Respiratory Symptom Survey, WURSS-21), and blood was obtained for biochemical analysis. Almost all athletes were in positive energy balance during OffS, while bodybuilders presented markedly restricted energy intake (â¼45%) leading to loss of weight (-9%) and fat mass (-45%) with preservation of fat-free mass in PreC. Protein intake was high during both periods, while lipid and carbohydrate intakes were reduced â¼50% in PreC. Almost all athletes consumed 100% of the Recommended Dietary Allowance (RDA) for micronutrients in OffS, while 45% and 75% of the athletes had intakes below the RDA for vitamins A and E in PreC. Oxidative damage to lipids (thiobarbituric acid reactive substances, TBARS), protein carbonyls, and the TBARS/total antioxidant capacity ratio increased in PreC (32%, 27%, 60%), as did plasma tumor necrosis factor α (4-fold) and WURSS-21 scores (25%). There were no significant changes in serum catalase, glutathione reductase, and superoxide dismutase activities nor in interleukin 1ß and immunoglobulins. In PreC, POMS showed negative changes in vigor (-20%), fatigue (23%), and total mood disturbance (35%), and RESTQ-Sport showed alterations for general and sport stress (34% and 50%, respectively) and sport recovery (-23%). Thus, PreC negatively affects nutrient intake, which may worsen oxidative stress, inflammation, psychological status, and the severity of respiratory infections in bodybuilders.
Asunto(s)
Atletas , Inflamación/fisiopatología , Estrés Oxidativo , Infecciones del Sistema Respiratorio/fisiopatología , Fenómenos Fisiológicos en la Nutrición Deportiva , Afecto , Antioxidantes/metabolismo , Atletas/psicología , Conducta Competitiva , Dieta , Ingestión de Energía , Metabolismo Energético , Humanos , Peroxidación de Lípido , Masculino , Ingesta Diaria Recomendada , Estaciones del Año , Deportes , Sustancias Reactivas al Ácido Tiobarbitúrico/metabolismoRESUMEN
We tested whether aerobic exercise training (AET) would modulate the skeletal muscle protein quality control (PQC) in a model of chronic kidney disease (CKD) in rats. Adult Wistar rats were evaluated in four groups: control (CS) or trained (CE), and 5/6 nephrectomy sedentary (5/6NxS) or trained (5/6NxE). Exercised rats were submitted to treadmill exercise (60 min., five times/wk for 2 months). We evaluated motor performance (tolerance to exercise on the treadmill and rotarod), cross-sectional area (CSA), gene and protein levels related to the unfolded protein response (UPR), protein synthesis/survive and apoptosis signalling, accumulated misfolded proteins, chymotrypsin-like proteasome activity (UPS activity), redox balance and heat-shock protein (HSP) levels in the tibialis anterior. 5/6NxS presented a trend towards to atrophy, with a reduction in motor performance, down-regulation of protein synthesis and up-regulation of apoptosis signalling; increases in UPS activity, misfolded proteins, GRP78, derlin, HSP27 and HSP70 protein levels, ATF4 and GRP78 genes; and increase in oxidative damage compared to CS group. In 5/6NxE, we observed a restoration in exercise tolerance, accumulated misfolded proteins, UPS activity, protein synthesis/apoptosis signalling, derlin, HSPs protein levels as well as increase in ATF4, GRP78 genes and ATF6α protein levels accompanied by a decrease in oxidative damage and increased catalase and glutathione peroxidase activities. The results suggest a disruption of PQC in white muscle fibres of CKD rats previous to the atrophy. AET can rescue this disruption for the UPR, prevent accumulated misfolded proteins and reduce oxidative damage, HSPs protein levels and exercise tolerance.
Asunto(s)
Actividad Motora/fisiología , Atrofia Muscular/prevención & control , Condicionamiento Físico Animal , Biosíntesis de Proteínas , Insuficiencia Renal Crónica/terapia , Factor de Transcripción Activador 4/genética , Factor de Transcripción Activador 4/metabolismo , Factor de Transcripción Activador 6/genética , Factor de Transcripción Activador 6/metabolismo , Animales , Catalasa/genética , Catalasa/metabolismo , Modelos Animales de Enfermedad , Regulación de la Expresión Génica , Glutatión Peroxidasa/genética , Glutatión Peroxidasa/metabolismo , Proteínas de Choque Térmico/genética , Proteínas de Choque Térmico/metabolismo , Pruebas de Función Renal , Masculino , Músculo Esquelético/metabolismo , Músculo Esquelético/fisiopatología , Atrofia Muscular/genética , Atrofia Muscular/metabolismo , Atrofia Muscular/fisiopatología , Nefrectomía/métodos , Complejo de la Endopetidasa Proteasomal/metabolismo , Ratas , Ratas Wistar , Insuficiencia Renal Crónica/genética , Insuficiencia Renal Crónica/fisiopatología , Insuficiencia Renal Crónica/cirugía , Prueba de Desempeño de Rotación con Aceleración Constante , Conducta Sedentaria , Transducción de SeñalRESUMEN
OBJECTIVE:: We aimed to determine whether aerobic training decreases superoxide levels, increases nitric oxide levels, and improves endothelium-dependent vasodilation in the aortas of spontaneously hypertensive rats. METHODS:: Spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) were distributed into 2 groups: sedentary (SHRsd and WKYsd, n=10 each) and swimming-trained (SHRtr, n=10 and WKYtr, n=10, respectively). The trained group participated in training sessions 5 days/week for 1 h/day with an additional work load of 4% of the animal's body weight. After a 10-week sedentary or aerobic training period, the rats were euthanized. The thoracic aortas were removed to evaluate the vasodilator response to acetylcholine (10-10 to 10-4 M) with or without preincubation with L-NG-nitro-L-arginine methyl ester hydrochloride (L-NAME; 10-4 M) in vitro. The aortic tissue was also used to assess the levels of the endothelial nitric oxide synthase and nicotinamide adenine dinucleotide oxidase subunit isoforms 1 and 4 proteins, as well as the superoxide and nitrite contents. Blood pressure was measured using a computerized tail-cuff system. RESULTS:: Aerobic training significantly increased the acetylcholine-induced maximum vasodilation observed in the SHRtr group compared with the SHRsd group (85.9±4.3 vs. 71.6±5.2%). Additionally, in the SHRtr group, superoxide levels were significantly decreased, nitric oxide bioavailability was improved, and the levels of the nicotinamide adenine dinucleotide oxidase subunit isoform 4 protein were decreased compared to the SHRsd group. Moreover, after training, the blood pressure of the SHRtr group decreased compared to the SHRsd group. Exercise training had no effect on the blood pressure of the WKYtr group. CONCLUSIONS:: In SHR, aerobic swim training decreased vascular superoxide generation by nicotinamide adenine dinucleotide oxidase subunit isoform 4 and increased nitric oxide bioavailability, thereby improving endothelial function.
Asunto(s)
Aorta Torácica/fisiopatología , Endotelio Vascular/fisiopatología , Hipertensión/fisiopatología , Condicionamiento Físico Animal/fisiología , Superóxidos/análisis , Natación/fisiología , Animales , Western Blotting , Etidio/análogos & derivados , Prueba de Esfuerzo , Fluorescencia , Hemodinámica , Masculino , NAD/análisis , NG-Nitroarginina Metil Éster/análisis , NG-Nitroarginina Metil Éster/metabolismo , Óxido Nítrico Sintasa de Tipo III/análisis , Óxido Nítrico Sintasa de Tipo III/metabolismo , Nitritos/análisis , Nitritos/metabolismo , Distribución Aleatoria , Ratas Endogámicas SHR , Valores de Referencia , Reproducibilidad de los Resultados , Superóxidos/metabolismo , Factores de Tiempo , Vasodilatación/fisiologíaRESUMEN
OBJECTIVE: We aimed to determine whether aerobic training decreases superoxide levels, increases nitric oxide levels, and improves endothelium-dependent vasodilation in the aortas of spontaneously hypertensive rats. METHODS: Spontaneously hypertensive rats (SHR) and Wistar Kyoto rats (WKY) were distributed into 2 groups: sedentary (SHRsd and WKYsd, n=10 each) and swimming-trained (SHRtr, n=10 and WKYtr, n=10, respectively). The trained group participated in training sessions 5 days/week for 1 h/day with an additional work load of 4% of the animal’s body weight. After a 10-week sedentary or aerobic training period, the rats were euthanized. The thoracic aortas were removed to evaluate the vasodilator response to acetylcholine (10-10 to 10-4 M) with or without preincubation with L-NG-nitro-L-arginine methyl ester hydrochloride (L-NAME; 10-4 M) in vitro. The aortic tissue was also used to assess the levels of the endothelial nitric oxide synthase and nicotinamide adenine dinucleotide oxidase subunit isoforms 1 and 4 proteins, as well as the superoxide and nitrite contents. Blood pressure was measured using a computerized tail-cuff system. RESULTS: Aerobic training significantly increased the acetylcholine-induced maximum vasodilation observed in the SHRtr group compared with the SHRsd group (85.9±4.3 vs. 71.6±5.2%). Additionally, in the SHRtr group, superoxide levels were significantly decreased, nitric oxide bioavailability was improved, and the levels of the nicotinamide adenine dinucleotide oxidase subunit isoform 4 protein were decreased compared to the SHRsd group. Moreover, after training, the blood pressure of the SHRtr group decreased compared to the SHRsd group. Exercise training had no effect on the blood pressure of the WKYtr group. CONCLUSIONS: In SHR, aerobic swim training decreased vascular superoxide generation by nicotinamide adenine dinucleotide oxidase subunit isoform 4 and increased nitric oxide bioavailability, thereby improving endothelial function.
Asunto(s)
Animales , Masculino , Aorta Torácica/fisiopatología , Endotelio Vascular/fisiopatología , Hipertensión/fisiopatología , Condicionamiento Físico Animal/fisiología , Superóxidos/análisis , Natación/fisiología , Western Blotting , Etidio/análogos & derivados , Prueba de Esfuerzo , Fluorescencia , Hemodinámica , NAD/análisis , NG-Nitroarginina Metil Éster/análisis , NG-Nitroarginina Metil Éster/metabolismo , Óxido Nítrico Sintasa de Tipo III/análisis , Óxido Nítrico Sintasa de Tipo III/metabolismo , Nitritos/análisis , Nitritos/metabolismo , Distribución Aleatoria , Ratas Endogámicas SHR , Valores de Referencia , Reproducibilidad de los Resultados , Superóxidos/metabolismo , Factores de Tiempo , Vasodilatación/fisiologíaRESUMEN
The effects of either eicosapentaenoic (EPA)- or docosahexaenoic (DHA)-rich fish oils on hindlimb suspension (HS)-induced muscle disuse atrophy were compared. Daily oral supplementations (0.3 mL/100 g b.w.) with mineral oil (MO) or high EPA or high DHA fish oils were performed in adult rats. After 2 weeks, the animals were subjected to HS for further 2 weeks. The treatments were maintained alongside HS At the end of 4 weeks, we evaluated: body weight gain, muscle mass and fat depots, composition of fatty acids, cross-sectional areas (CSA) of the soleus muscle and soleus muscle fibers, activities of cathepsin L and 26S proteasome, and content of carbonylated proteins in the soleus muscle. Signaling pathway activities associated with protein synthesis (Akt, p70S6K, S6, 4EBP1, and GSK3-beta) and protein degradation (atrogin-1/MAFbx, and MuRF1) were evaluated. HS decreased muscle mass, CSA of soleus muscle and soleus muscle fibers, and altered signaling associated with protein synthesis (decreased) and protein degradation (increased). The treatment with either fish oil decreased the ratio of omega-6/omega-3 fatty acids and changed protein synthesis-associated signaling. EPA-rich fish oil attenuated the changes induced by HS on 26S proteasome activity, CSA of soleus muscle fibers, and levels of p-Akt, total p70S6K, p-p70S6K/total p70S6K, p-4EBP1, p-GSK3-beta, p-ERK2, and total ERK 1/2 proteins. DHA-rich fish oil attenuated the changes induced by HS on p-4EBP1 and total ERK1 levels. The effects of EPA-rich fish oil on protein synthesis signaling were more pronounced. Both EPA- and DHA-rich fish oils did not impact skeletal muscle mass loss induced by non-inflammatory HS.
Asunto(s)
Ácidos Docosahexaenoicos/administración & dosificación , Ácido Eicosapentaenoico/administración & dosificación , Aceites de Pescado/química , Redes Reguladoras de Genes , Suspensión Trasera/efectos adversos , Trastornos Musculares Atróficos/metabolismo , Animales , Ácidos Docosahexaenoicos/farmacología , Ácido Eicosapentaenoico/farmacología , Redes Reguladoras de Genes/efectos de los fármacos , Masculino , Músculo Esquelético/efectos de los fármacos , Trastornos Musculares Atróficos/etiología , Ratas , Transducción de Señal/efectos de los fármacosRESUMEN
Age-related loss of muscle mass and strength can be accelerated by impaired recovery of muscle mass following a transient atrophic stimulus. The aim of this study was to identify the mechanisms underlying the attenuated recovery of muscle mass and strength in old rats following disuse-induced atrophy. Adult (9 month) and old (29 month) male F344BN rats underwent hindlimb unloading (HU) followed by reloading. HU induced significant atrophy of the hindlimb muscles in both adult (17-38%) and old (8-29%) rats, but only the adult rats exhibited full recovery of muscle mass and strength upon reloading. Upon reloading, total RNA and protein synthesis increased to a similar extent in adult and old muscles. At baseline and upon reloading, however, proteasome-mediated degradation was suppressed leading to an accumulation of ubiquitin-tagged proteins and p62. Further, ER stress, as measured by CHOP expression, was elevated at baseline and upon reloading in old rats. Analysis of mRNA expression revealed increases in HDAC4, Runx1, myogenin, Gadd45a, and the AChRs in old rats, suggesting neuromuscular junction instability/denervation. Collectively, our data suggests that with aging, impaired neuromuscular transmission and deficits in the proteostasis network contribute to defects in muscle fiber remodeling and functional recovery of muscle mass and strength.
Asunto(s)
Envejecimiento/metabolismo , Estrés del Retículo Endoplásmico , Retículo Endoplásmico/metabolismo , Proteínas Musculares/biosíntesis , Fuerza Muscular , Músculo Esquelético/metabolismo , Atrofia Muscular/metabolismo , Factores de Edad , Envejecimiento/genética , Envejecimiento/patología , Animales , Autofagia , Modelos Animales de Enfermedad , Retículo Endoplásmico/patología , Regulación de la Expresión Génica , Suspensión Trasera , Masculino , Proteínas Musculares/genética , Músculo Esquelético/patología , Músculo Esquelético/fisiopatología , Atrofia Muscular/genética , Atrofia Muscular/patología , Atrofia Muscular/fisiopatología , Complejo de la Endopetidasa Proteasomal/metabolismo , Biosíntesis de Proteínas , Proteolisis , Ratas Endogámicas F344 , Recuperación de la Función , Factores de TiempoRESUMEN
This study aimed at investigating the acute effects of aerobic exercise on endothelium-dependent vasomotor function of rat aorta, as well as mechanisms involved in endothelial nitric oxide (NO) bioactivity. Wistar rats were assigned to either a resting control (C, n = 21) or acutely exercised (E, n = 21) groups (60 min, 55-60% of maximum speed). After exercise, thoracic aorta was excised and cut into rings. Two rings were promptly applied to evaluate vasomotor function and the rest of aorta was used for additional measurements. Acute exercise significantly improved maximum ACh-induced relaxation (C, 91.6 ± 1.2 vs. E, 102.4 ± 1.7%, p < 0.001) and sensitivity to ACh (C, -7.3 ± 0.06 vs. E, -7.3 ± 0.02 log M, p < 0.01), and was accompanied by significantly increases on serine1177 eNOS phosphorylation, reflecting its enhanced activation. However, acute exercise also enhanced both superoxide and hydrogen peroxide production, as assayed by dihydroethidium oxidation, lucigenin chemiluminescence and Amplex Red assays. We also provided evidence for Nox2 NADPH oxidase (Nox) activation through gp91dstat-mediated inhibition of superoxide signals. Enhanced arterial relaxations associated with acute exercise were nearly-completely prevented by catalase, suggesting a role for paracrine hydrogen peroxide. Despite increased detectable oxidant generation, cellular oxidative stress was not evident, as suggested by unaltered GSH:GSSG ratio and lipid hydroperoxides. Collectively, these results demonstrate that one bout of moderate aerobic exercise improves endothelial function by increasing NO bioavailability, while superoxide and hydrogen peroxide are generated in a controlled fashion.
Asunto(s)
Endotelio Vascular/metabolismo , Óxido Nítrico/metabolismo , Condicionamiento Físico Animal/fisiología , Especies Reactivas de Oxígeno/metabolismo , Vasodilatación/fisiología , Acetilcolina/metabolismo , Animales , Aorta/química , Aorta/metabolismo , Masculino , Óxido Nítrico/análisis , Ratas , Ratas Wistar , Especies Reactivas de Oxígeno/análisisRESUMEN
Excess of saturated fatty acids in the diet has been associated with obesity, leading to systemic disruption of insulin signaling, glucose intolerance, and inflammation. Macadamia oil administration has been shown to improve lipid profile in humans. We evaluated the effect of macadamia oil supplementation on insulin sensitivity, inflammation, lipid profile, and adipocyte size in high-fat diet (HF) induced obesity in mice. C57BL/6 male mice (8 weeks) were divided into four groups: (a) control diet (CD), (b) HF, (c) CD supplemented with macadamia oil by gavage at 2 g/Kg of body weight, three times per week, for 12 weeks (CD + MO), and (d) HF diet supplemented with macadamia oil (HF + MO). CD and HF mice were supplemented with water. HF mice showed hypercholesterolemia and decreased insulin sensitivity as also previously shown. HF induced inflammation in adipose tissue and peritoneal macrophages, as well as adipocyte hypertrophy. Macadamia oil supplementation attenuated hypertrophy of adipocytes and inflammation in the adipose tissue and macrophages.
Asunto(s)
Inflamación/dietoterapia , Macadamia , Obesidad/dietoterapia , Aceites de Plantas/administración & dosificación , Adipocitos/patología , Animales , Aumento de la Célula , Colesterol/sangre , Citocinas/biosíntesis , Dieta Alta en Grasa/efectos adversos , Inflamación/metabolismo , Inflamación/patología , Resistencia a la Insulina , Macrófagos Peritoneales/metabolismo , Macrófagos Peritoneales/patología , Masculino , Ratones , Ratones Endogámicos C57BL , Óxido Nítrico/metabolismo , Obesidad/metabolismo , Obesidad/patologíaRESUMEN
The effects of adipose-derived mesenchymal stem cells (ADMSC) transplantation on degeneration, regeneration and skeletal muscle function were investigated in dystrophin-deficient mice (24-week-old). ADMSC transplantation improved muscle strength and, resistance to fatigue. An increase in fiber cross-sectional area and in the number of fibers with centralized nuclei and augment of myogenin content were observed. In ADMSC-treated muscles a decrease in muscle content of TNF-α, IL-6 and oxidative stress measured by Amplex(®) reagent were observed. The level of TGF-ß1 was lowered whereas that of VEGF, IL-10 and IL-4 were increased by ADMSC treatment. An increase in markers of macrophage M1 (CD11 and F4-80) and a decrease in T lymphocyte marker (CD3) and arginase-1 were also observed in ADMSCs-treated dystrophic muscle. No change was observed in iNOS expression. Increased phosphorylation of Akt, p70S6k and 4E-BP1 was found in dystrophic muscles treated with ADMSC. These results suggest that ADMSC transplantation modulates inflammation and improves muscle tissue regeneration, ameliorating the dystrophic phenotype in dystrophin-deficient mice.