Proteomic changes in skeletal muscle of aged rats in response to resistance training.

Sarcopenia is a multifactorial process defined by loss of strength and skeletal muscle mass, which leads to a reduction in muscle cross-sectional area (CSA). Although resistance training (RT) has been indicated as a tool to counteract sarcopenia, the protein profile associated with skeletal muscle adaptations remains to be determined. We investigated the effects of 12 weeks of RT on the skeletal muscle proteome profile and CSA of young and older rats. Twenty-four animals were divided into four groups: young sedentary or trained and older sedentary or trained (six animals per group). A 12-week RT protocol was performed, which consisted of climbing a vertical ladder. The proteins from the gastrocnemius were analysed by LC-ESI-MS/MS. One-hundred and thirty-one proteins were identified, of which 28 were assessed between the groups. Ageing induced an increase in proteins associated with the glycolytic pathway, transport and stress response, which represent crucial mechanisms for muscle adaptation. RT upregulated metabolic enzymes, anti-oxidant activity and transport proteins, besides increasing hypertrophy, regardless of age, suggesting a beneficial adaptation to mitigate age-related sarcopenia. RT reduced muscle atrophy through the regulation of stress response and by increasing proteins related to energy production and transport, which in turn might protect tissue damage arising from exercise and ageing. SIGNIFICANCE OF THE STUDY: Protein abundance levels related to the metabolic process and stress response were increased in the aged muscle. RT proved to be an important intervention capable of inducing significant effects on muscle proteome regardless of ageing, due to upregulation of glycolytic enzymes, and anti-oxidant and transport proteins. This effect could lead to a beneficial adaptation in muscle structure, cellular function and overall homeostasis maintenance. This study contributes to better understanding of the basic biology of ageing and clarifies more profoundly the molecular networks behind physiological adaptations promoted by exercise training. Therefore, the results open new perspectives and insights for studies based on transcriptomics, metabolomics and functional assays.

Resistance training improves body composition and increases matrix metalloproteinase 2 activity in biceps and gastrocnemius muscles of diet-induced obese rats

OBJECTIVE: We investigated the influence of resistance training on body composition and matrix metalloproteinase 2 activity in skeletal muscles of rats fed a high-fat diet. METHODS: Thirty-two Wistar rats were divided into four experimental groups (n = 8/each) according to diet and exercise status: Control (standard diet), Obese Control (high-fat diet), Resistance Training (standard diet) and Obese Resistance Training (high-fat diet) groups. Animals were fed a high-fat diet for 12 weeks to promote excessive weight gain. Resistance Training groups performed 12 weeks of training periods after this period in a vertical ladder three times/week. Fat percentage, fat-free mass and fat mass were assessed using dual-energy X-ray absorptiometry, and matrix metalloproteinase 2 activity in biceps and gastrocnemius muscles was analyzed using zymography. RESULTS: Resistance training significantly reduced body and fat masses and fat percentages in both trained groups (p<0.05). The maximal carrying load between trained groups was not different, but relative force was higher in the Resistance Training group (p<0.05). Of note, increased matrix metalloproteinase 2 activity was noted in the tested muscles of both trained groups (p<0.05). CONCLUSION: In conclusion, altered body composition and muscle matrix metalloproteinase 2 activity promoted by excessive weight gain were positively modified by resistance training. .

Low-level laser therapy (LLLT) (660nm) alters gene expression during muscle healing in rats.

INTRODUCTION: The effects of LLLT were studied during muscle regeneration through gene expression. METHODS: It was evaluated 10 and 50J/cm(2) doses during 7, 14 and 21days post cryoinjury, through histopathological analysis and mRNA MyoD, Myogenin, Vegf and Cox-2 expression. RESULTS: Irradiated groups presented less inflammatory process than control group after 14 and 21days. Cox-2 levels were downregulated in all irradiated groups after 7, 14 and 21days. On day 7, both treated groups had a downregulation of Vegf levels, and an upregulation after 14 and 21days, mainly with 50J/cm(2). The MyoD levels were upregulated with high dose in all periods and with low dose after 21days. Myogenin expression was downregulated in both treated groups after 7days, and was upregulated with 10J/cm(2) after 21days. CONCLUSION: These responses suggest that LLLT can improve the skeletal muscle regeneration through the gene expression stimulation.

Biomarker responses as indication of contaminant effects in Oreochromis niloticus.

The current study investigated oxidative stress parameters (enzymes activities, metallothionein content and lipid peroxidation) in freshwater fish, Oreochromis niloticus, tilapia exposure to Monjolinho River (in 4 months of year: January, April, July and November). One critical site in Monjolinho River (site B) was assessed in comparison to a reference site (site A). Water pH and oxygen concentration was lower than that recommended by CONAMA (Brazilian National Environmental Committee), resolution 357/2005 for protection of aquatic communities, and ammonium and the metals Cu, Zn, Mn and Fe (on all months) concentrations were higher than the maximum concentration recommended. Glutathione peroxidase (GPx) and superoxide dismutase (SOD) activities were significantly decreased in liver and muscle in tilapia from Monjolinho River, throughout the year, in relation to reference except in gills that SOD activity increased. Glutathione S-transferase (GST) activity was significantly increased in liver of the tilapia from Monjolinho River in all sites, in relation to reference except in gills that GST activity increased in July and decreased in November, suggesting that GST activity could be induced to neutralize the pollutants toxicity. On the other hand, GST activity was significantly decreased in white muscle indicating a toxic effect of pollutants, resulting in a decreased ability of tilapia to perform defense reactions associated to GSTs. The decrease of catalase (CAT) activity in gills of the O. niloticus together with the increase of SOD activity, could explain the increased lipid peroxidation (LPO) level in this organ. Metallothionein levels in liver and gills were significantly high in all sites. Results indicate that the exposure to metals caused severe damage to tissues; despite the consensually assumed antioxidant induction as a sign of exposure to contaminants the effects seem in part to be mediated by suppression of antioxidant system with SOD, CAT and GPx as potential candidates for tissues toxicity biomarkers of pollutants.

Resistance training restores the gene expression of molecules related to fat oxidation and lipogenesis in the liver of ovariectomized rats.

Decreased levels of estrogen are associated with hepatic steatosis (HS), through changes in gene expression of molecules related to fat oxidation and lipogenesis. Both resistance training (RT) and endurance training (ET) prevent HS in ovariectomized (Ovx) rats. However, the molecular events associated with this process were only investigated for ET, but not for RT. Thus, the aim of this study was to investigate the effects of Ovx and RT on the gene expression of molecules related to fat oxidation and lipogenesis in the liver of rats. Sprague-Dawley adult female rats were grouped into four (n = 6 per group): sham-operated sedentary (Sham-Sed); Ovx sedentary (Ovx-Sed); sham-Rt and Ovx-Rt. A 10-week RT period, during which the animals climbed a 1.1-m vertical ladder with weights attached to their tails, was used. The sessions were performed three times a week, with 4-9 climbs and 8-12 dynamic movements per climb. Gene expression was analyzed by RT-PCR by the ∆∆Ct method. The estrogen deficiency associated with ovariectomy decreased the gene expression of molecules related to fat oxidation, carnitine palmitoyltransferase I (53%) and ß-hydroxyacyl-CoA dehydrogenase (27%), and increased molecules related to lipogenesis, sterol regulatory element-binding protein-1c (106%), acetyl-CoA carboxylase (ACC) (72%) and stearoyl CoA desaturase-1 (109%). With the exception of ACC, the ovariectomy-induced changes in the expression of these molecules were restored by RT. The present results indicate that RT has important effects on the prevention of HS in Ovx animals, through changes in gene expression of molecules related to hepatic lipid metabolism.

Low-level laser therapy induces differential expression of osteogenic genes during bone repair in rats.

OBJECTIVES: The aim of this study was to measure the temporal pattern of the expression of osteogenic genes after low-level laser therapy during the process of bone healing. We used quantitative real-time polymerase chain reaction (qPCR) along with histology to assess gene expression following laser irradiation on created bone defects in tibias of rats. MATERIAL AND METHODS: The animals were randomly distributed into two groups: control or laser-irradiated group. Noncritical size bone defects were surgically created at the upper third of the tibia. Laser irradiation started 24 h post-surgery and was performed for 3, 6, and 12 sessions, with an interval of 48 h. A 830 nm laser, 50 J/cm(2), 30 mW, was used. On days 7, 13, and 25 post-injury, rats were sacrificed individually by carbon dioxide asphyxia. The tibias were removed for analysis. RESULTS: The histological results revealed intense new bone formation surrounded by highly vascularized connective tissue presenting slight osteogenic activity, with primary bone deposition in the group exposed to laser in the intermediary (13 days) and late stages of repair (25 days). The quantitative real-time PCR showed that laser irradiation produced an upregulation of BMP-4 at day 13 post-surgery and an upregulation of BMP4, ALP, and Runx 2 at day 25 after surgery. CONCLUSION: Our results indicate that laser therapy improves bone repair in rats as depicted by differential histopathological and osteogenic genes expression, mainly at the late stages of recovery.

Oxidative stress biomarkers of exposure in the blood of cichlid species from a metal-contaminated river.

The oxidative stress biomarkers of exposure, such as reduced glutathione (GSH), activity of superoxide dismutase (SOD), catalase (CAT) and glutathione peroxidase (GPx) and the levels of lipid peroxidation (LPO), were measured in the blood of three cichlid fish (Oreochromis niloticus, Tilapia rendalli, and Geophagus brasiliensis) taken during two seasons from two sites, unpolluted and polluted by industrial effluents, to evaluate the effectiveness of these biomarkers in assessing the impact of water contamination. The LPO levels in the blood were higher in fish from the metal-contaminated site and the chronic exposure led to significant changes in GPx, CAT, and SOD activities in all three cichlid species. The considerable variation of responses in these cichlids to water contamination evidenced differences in sensitivity to the metal contamination and/or in the potential to respond to it highlighting the importance of using a set of related biomarkers to assess the impact of water contamination.

Hepatic metallothionein in a teleost (Prochilodus scrofa) exposed to copper at pH 4.5 and pH 8.0.

Copper accumulation and induction of metallothionein (MT) were investigated in the liver of juvenile Prochilodus scrofa exposed to copper (96 h-LC50) at pH 4.5 (LC50=98+/-0.8 microg Cu l(-1)) and pH 8.0 (LC50=16+/-0.5 microg Cu l(-1)) at 20 degrees C. No significant difference was found in copper accumulated in the liver at either pH. The ratio between copper accumulation in the liver after 96 h exposure and the copper in the water at pH 4.5 and 8.0 was 5.1 and 34.4, respectively. MT in the liver of fish exposed to copper was isolated by DEAE-Sepharose chromatography and identified by SDS-PAGE. No MT was identified in the controls. MT concentration was 126.8+/-21.5 and 167.4+/-25.6 microg g(-1) wet mass in fish exposed to copper at pH 4.5 and pH 8.0 (P>0.05), respectively. The results indicate that the pH of the water is a determining factor in copper toxicity, and the increased concentration of MT in the liver of P. scrofa exposed to copper suggests the possibility of using MT as a biomarker to evaluate the biological response of this species to copper contamination in aquatic environments.