Fluorescence was normalized to complete protein content material, and expressed as AU per mg of protein All info had been analysed using a 2 (WKY vs. SHR) x two (SED vs. EX) 92831-11-3 factorial ANOVA. Tukey’s publish-hoc check was used the place suitable. In a one instance (WG 685898-44-6 calpain activity) a Student’s T-take a look at was utilized to analyse differences between the SHRSED and SHREX teams. Statistical significance was regarded as at p<0.05.The activity of the apoptosis-related enzyme caspase-3 was higher (p<0.005) in the WG of SHR compared to WKY rats, but was significantly (p<0.001) lower with training. The activity of the lysosomal enzyme cathepsin was elevated (p<0.05) in the WG of the SHRSED group compared to all groups, and significantly (p<0.001) reduced in SHREX animals. Calpain activity was elevated by 26.3% in the SHRSED compared to WKYSED group. T-test analysis revealed that calpain activity was significantly (p<0.05) reduced by 24.9% in exercise-trained compared to sedentary hypertensive animals (SHREX vs. SHRSED) (Fig. 1A). Caspase-3 activity was not significantly different between strains in the LV, but was significantly (p<0.001) lower with training. In the LV, cathepsin activity was elevated (p<0.001) in SHR compared to WKY animals, with exercise training tending (p = 0.09) to reduce cathepsin activity. Calpain activity was not different across strains in the LV however, exercise training reduced (p<0.001) LV calpain activity (Fig. 1A). ROS generation tended (p = 0.09) to be greater in the WG of SHR compared to WKY rats however, exercise did not alter WG ROS generation. In the LV, no difference was observed in ROS generation between SHR and WKY animals (Fig. 1B). However, exercise training tended to reduce (p = 0.08) LV ROS generation. MAFbx and MuRF1 protein content did not differ between strains and was not affected by exercise in both the WG and LV (Fig. 2A). However, proteasome activity in the WG was significantly higher (p<0.001) in SHR compared to WKY animals, and further elevated (p<0.001) with exercise (Fig. 2B). Proteasome activity was also elevated (p<0.05) with exercise training in the LV (Fig. 2B).To examine if the observed changes in proteolytic markers were related to autophagy, we measured several key factors indicative of autophagosome content. There was a trend (p = 0.07) for elevated LC3 mRNA in the WG of SHRSED compared to the WKYSED group. In addition, LC3 mRNA was significantly higher in the WKYEX (p<0.005) and SHREX (p<0.05) groups compared to the WKYSED group (Fig. 3A). p62 mRNA in the WG was not different across strains but was significantly (p<0.05) higher with exercise training (Fig. 3B). Higher levels of LAMP2 mRNA (p<0.05) were present in hypertensive WG muscle, but were not affected by Fig 1. Proteolytic enzyme activity and ROS generation in muscle of sedentary and exercise-trained normotensive and hypertensive rats. A: quantitative analysis of caspase-3, calpain, and cathepsin enzymatic activity in the white gastrocnemius (WG) and left ventricle (LV). B: quantitative analysis of ROS generation in the WG and LV. Values are means SEM (n = 92). p<0.001 vs WKY (main effect) p<0.005 vs WKY (main effect) p<0.001 vs SED (main effect) p<0.05 vs all groups (interaction effect) 1 p<0.001 vs all groups (interaction effect) p<0.05 vs. SHRSED (T-test). exercise (Fig. 3C). In the WG, a significant increase (p<0.005) in LC3I protein was found in hypertensive animals, but there was no change with exercise. LC3II protein did not differ in the WG across strains or by training status. The LC3II:I ratio was lower (p<0.005) in the WG of hypertensive rats, but was not affected by exercise (Fig. 4A). In the LV, LC3 and LAMP2 mRNA levels were not different across strains or exercise condition (Fig. 3A & C). p62 mRNA levels were not different across strains in the LV, but were significantly (p<0.05) higher with exercise (Fig. 3B).