SBP (P = 0.023 and P = 0.01, respectively) and increase in VO2 Max (P = 0.01) along with reduced inflammatory response as indicated by reduced levels of TNF-a and IL-6 (P = 0.004 and P = 0.012, respectively). Exercise triggered also a decrease in TBARS levels (P = 0.004). In order to assess the effect of exercise on the expression of DNAJB3, fat adipose tissue biopsies were collected from obese subjects that completed the exercise program and used to monitor the expression of DNAJB3 (n = 10). As shown in Figure 3A, there was a significant increase in the levels of DNAJB3 mRNA in obese subjects after exercise (P = 0.005). Consistent with the RT-PCR data, IHC analysis confirmed that DNAJB3 protein is significantly increased in the adipose tissue of obese subjects after physical exercise (Fig. 3B; P = 0.003). Taken together, these data suggest that exercise can interfere with obesity-mediated repression of DNAJB3. Since obesity is known to trigger the activation of the stress kinase JNK; particularly its phosphorylation, and with the perspective to establish a correlation between the levels of DNAJB3 and activated JNK, we measured the levels of phosphorylated JNK in adipose tissue by IHC before and after exercise. In contrast to the pattern observed for DNAJB3, there was a significant increase in the levels phosphorylated JNK in obese subjects compared to lean subjects (P = 0.Allantoin 016, data not shown). Furthermore, the expected increase in phosphorylated JNK in obese was significantly reduced by physical exercise (Fig. 3C; P = 0.0013). No effect was observed for total JNK before and after exercise (data not shown). From this experiment we concluded that there was an inverse correlation between DNAJB3 and activated JNK.Correlation analysis of DNAJB3 with physical and clinical profiles, inflammatory and metabolic stress markersTo understand the physiological consequence of the reduction of DNAJB3 in obese subjects on the clinical profile as well as the inflammatory and metabolic stress responses, we investigated their correlation with DNAJB3 mRNA levels using Spearman’s rank test before and after exercise.Fosaprepitant dimeglumine Table 5 shows that before exercise, there was a negative correlation between levels of DNAJB3 and the indicators of obesity such as BMI (r2 = 20.PMID:25558565 71; P,0.0001) and PBF (r2 = 20.66; P = 0.0001). There was also a negative correlation with TG (r2 = 20.36; P,0.035) as well as with the pro-inflammatory chemokines IP-10 and RANTES (r2 = 20.37; P,0.036 and r2 = 20.40; P = 0.02, respectively). No correlation was found for the other parameters (Table 5 and data not shown). After exercise, the increased expression of DNAJB3 mRNA in obese correlated negatively with the PBF (r2 = 20.53; P = 0.044) and positively with RANTES (r2 = 0.75; P,0.008) (Table 5).Validation of the transcriptomic data by Western Blotting and ImmunohistochemistryIn order to validate the RT-PCR data on these members of the Hsp-40 at the protein level, we performed Western blot analysis on PBMCs and immunohistochemical (IHC) analysis on adipose tissue from selected subjects. As shown in Figure 2A, Western blot analysis performed on lean and obese subjects (n = 4 for each group) showed a major reduction in the expression of DNAJB3 protein in obese subjects (P,0.05). Under the same conditions, we were unable to validate the expression of DNAJC5B and DNAJB7 (data not shown). In contrast to DNAJB3, the expression levels of HSC-70 and HSP-90 were increased in obese subjects (Fig. 2A). Consis.
