Relationship between NADP-specific isocitrate dehydrogenase and glutathione peroxidase in aging rat skeletal muscle.

The glutathione peroxidase (GPX) system detoxifies hydroperoxides in cells and uses NADPH to regenerate reduced glutathione. Enzymatic sources of NADPH in skeletal muscle include NADP-specific isocitrate dehydrogenase (ICDP), glucose-6-phosphate dehydrogenase (G6PD), and malic enzyme (ME). Our purpose was to explore the relationship in skeletal muscle between GPX and ICDP along with other NADPH-generating enzymes as a function of progressive age and muscle fiber-type. Soleus (SOL), red gastrocnemius (RG), and white gastrocnemius (WG) muscles were extracted from Fischer-344 rats of three different ages: 4 months old (Y); 18 months old (M); and 24 months old (O). Assays were conducted to determine activities of GPX, ICDP, G6PD, and ME along with levels of lipid hydroperoxides. GPX activities were significantly greater in RG and WG of old rats than in younger. ICDP activities were higher in the WG of old and middle aged rats when compared to young adults. GPX and ICDP activities exhibited similar differences among the muscles tested (SOL>RG>WG). In contrast, G6PD and ME activities were not significantly different across muscles. G6PD activities increased in RG with age, but were well over an order of magnitude lower than ICDP in all muscles. ME activities were universally lower than ICDP in all muscles, and decreased with old age in the WG and RG. Lipid hydroperoxides were significantly higher with aging in RG. Significant correlations were found between GPX and ICDP in all muscles. Stepwise regression resulted in a model (R(2)=0.82) that included ICDP and ME in predicting GPX. In summary, these data are consistent with the hypotheses that ICDP is higher in more oxidative fibers, inducible with aging, and most closely associated with the glutathione peroxidase system in skeletal muscle.

Ageing alters aortic antioxidant enzyme activities in Fischer-344 rats.

Oxidative stress imposed by reactive oxygen species is now believed to contribute to hypertension, atherosclerosis and ageing of the vasculature all involving a loss of relaxation. The antioxidant enzymes glutathione peroxidase, superoxide dismutase and catalase play a crucial role in defending against the ravages of oxidative stress. Our purpose was to characterize age-related changes in glutathione peroxidase, superoxide dismutase and catalase in the rat aorta. Aortas were extracted from seven young (4 months), seven middle aged (18 months) and seven old (24 months) animals. Analysis of variance was used with Fisher-LSD post hoc to determine mean differences among glutathione peroxidase, superoxide dismutase and catalase. Aortic glutathione peroxidase activities rose steadily with age expressed in micromol mg protein-1 min-1 +/- SEM (young: 141 +/- 22; middle aged: 198 +/- 18; old: 229 +/- 26) reaching significance between young and old. Superoxide dismutase activities significantly decreased in middle aged when compared with young (young: 22 +/- 2 vs. middle aged: 15 +/- 2 U mg protein-1) before trending upward again in old age (19 +/- 2). Catalase activities dropped significantly between young and old when expressed in mU mg protein-1 (young: 230 +/- 30; middle aged: 173 +/- 18; old: 144 +/- 23). Ratios for the various enzymes indicate a shrinking contribution of catalase with ageing, with an enhanced role for glutathione peroxidase in the antioxidant defence. These data in aortas of ageing rats show a complex alteration of the antioxidant profile.