Vitamin E and C supplementation reduces oxidative stress, improves antioxidant enzymes and positive muscle work in chronically loaded muscles of aged rats.

Aging is associated with increased oxidative stress. Muscle levels of oxidative stress are further elevated with exercise. The purpose of this study was to determine if dietary antioxidant supplementation would improve muscle function and cellular markers of oxidative stress in response to chronic repetitive loading in aging. The dorsiflexors of the left limb of aged and young adult Fischer 344 Brown×Norway rats were loaded 3 times weekly for 4.5 weeks using 80 maximal stretch-shortening contractions per session. The contra-lateral limb served as the intra-animal control. The rats were randomly assigned to a diet supplemented with Vitamin E and Vitamin C or normal non-supplemented rat chow. Biomarkers of oxidative stress were measured in the tibialis anterior muscle. Repetitive loading exercise increased maximal isometric force, negative work and positive work in the dorsiflexors of young adult rats. Only positive work increased in the aged animals that were supplemented with Vitamin E and C. Markers of oxidative stress (H(2)O(2), total GSH, GSH/GSSG ratio, malondialdehyde and 8-OHdG) increased in the tibialis anterior muscles from aged and young adult animals with repetitive loading, but Vitamin E and C supplements attenuated this increase. MnSOD activity increased with supplementation in the young adult animals. CuZnSOD and catalase activity increased with supplementation in young adult and aged animals and GPx activity increased with exercise in the non-supplemented young adult and aged animals. The increased levels of endogenous antioxidant enzymes after Vitamin E and C supplementation appear to be regulated by post-transcriptional modifications that are affected differently by age, exercise, and supplementation. These data suggest that antioxidant supplementation improves indices of oxidative stress associated with repetitive loading exercise and aging and improves the positive work output of muscles in aged rodents.

Aging-dependent regulation of antioxidant enzymes and redox status in chronically loaded rat dorsiflexor muscles.

This study compares changes in the pro-oxidant production and buffering capacity in young and aged skeletal muscle after exposure to chronic repetitive loading (RL). The dorsiflexors from one limb of young and aged rats were loaded 3 times/week for 4.5 weeks using 80 maximal stretch-shortening contractions per session. RL increased H2O2 in tibialis anterior muscles of young and aged rats and decreased the ratio of reduced/oxidized glutathione and lipid peroxidation in aged but not young adult animals. Glutathione peroxidase (GPx) activity decreased whereas catalase activity increased with RL in muscles from young and aged rats. RL increased CuZn superoxide disumutase (SOD) and Mn SOD protein concentration and CuZn SOD activity in muscles from young but not aged animals. There were no changes in protein content for GPx-1 and catalase or messenger RNA for any of the enzymes studied. These data show that aging reduces the adaptive capacity of muscles to buffer increased pro-oxidants imposed by chronic RL.

Chronic exposure to stretch-shortening contractions results in skeletal muscle adaptation in young rats and maladaptation in old rats.

The objective of this research was to investigate skeletal muscle response to a chronic administration of stretch-shortening cycles (SSCs) in young and old rats. Dorsiflexor muscles of old (30 months, n = 5) and young (12 weeks, n = 6) rats were exposed 3 times/week for 4.5 weeks to a protocol of 80 maximal SSCs per exposure in vivo. Skeletal muscle response was characterized by isometric and dynamic performance, as well as by muscle wet mass and quantitative morphological analyses following the exposure period. The performance of the young and old groups was not statistically different at the start of the exposure. By the end of the exposure, however, a statistical difference was noted, as performance increased significantly in the young animals and decreased significantly in the old animals. Muscle wet mass of the left tibialis anterior (TA) in the treated limb was significantly greater in the youngthan in the old animals (p < 0.001), whereas there was no difference in the contra-lateral TA. No degenerative myofibers or changes in non-cellular interstitium were noted in either age group, but a significant increase was observed in the volume of the cellular interstitium in the exposed limb of the old animals (p = 0.01), which is indicative of an inflammatory response. Thus, a chronic exposure of SSCs results in significant performance increase and muscle hypertrophy in young animals, and a significant performance decrease and an increased cellular interstitial response in old animals. These findings suggest that age may impair the ability of skeletal muscle to adapt to repetitive mechanical loading, even in the absence of degeneration.

Resistance training increases heat shock protein levels in skeletal muscle of young and old rats.

Heat shock proteins (HSP) HSP72, HSC70 and HSP25 protein levels and mRNA levels of HSP72 genes (Hsp72-1, Hsp72-2, Hsp72-3) and HSC70 were examined in tibialis anterior muscles from young and old rats following 4.5 weeks of heavy resistance exercise. Young (3 months) (n=10) and old (30 months) (n=9) rats were subjected to 14 sessions of electrically evoked resistance training using stretch-shortening contractions of the left limb that activated the dorsiflexor muscle group, including the tibialis anterior muscle, while the right side served as the intra-animal control. Muscle wet weight of the left tibialis anterior increased by 15.6% in young animals compared to the untrained right side, while the aged rats demonstrated no significant hypertrophy based on muscle wet weight. There were no differences in mRNA expression between the control and experimental muscles in either the old or the young animals for any of the four genes examined. On the other hand, HSP72 levels as determined by Western blots were significantly (p<0.01) higher (968.8 and 409.1%) in the trained as compared to the contralateral control muscle in young and old animals, respectively. HSP25 expression was increased significantly (p<0.01) by training in muscles of young rats (943.1%) and old rats (420.3%). Moreover, there was no training by age interaction for HSP72, while a significant age and training by age effects were found in muscles for HSP25. There was no change in HSC70 protein expression in response to the training intervention in either age group. SOD-1 enzyme level increased by 66.6% in the trained muscles of the young rats, while this enzyme was 33% lower in trained muscles compared to the untrained control side in old rats. Moreover, a significant (p<0.05) training by age interaction was found for SOD-1 enzyme levels. This study suggests that fast contracting muscles in young and old animals are capable of increasing HSP expression in response to high intensity contractile stress. Furthermore, the data are consistent with the hypothesis that higher levels of oxidative stress in muscles of old animals limit HSP levels and/or function in response to high intensity contractile stress.