001–1000 ng, Sigma–Aldrich, USA) were constructed by applying

in bolus injections (100 μl) in the coronary bed at 10 min intervals. After decapitation, blood samples were collected in sterile tubes containing EDTA/K3, centrifuged at 3000 × g for 15 min at 4 °C (Fanem, São Paulo, Brazil) and stored at −80 °C until use. Plasma 17β-estradiol concentrations were analyzed by an electrochemiluminescence immunoassay method (Elecsys 2010, Roche, Basel, Switzerland), with available kits (Estradiol II, Roche, Mannheim, Germany). The measures for right and left retroperitoneal abdominal adiposity (RET) and perirenal (PR), parametrial (PME), and inguinal (ING) adiposities were determined by means of bilateral lipectomy (the surgical extraction of fat pads). A longitudinal incision of ±6 cm was made on the abdominal skin using the

Alba line as a reference. Next, the ING compartments were mechanically collected and measured and Everolimus research buy the peritoneum was cut open, and the RET, PR and PME fat pads were taken out following a similar protocol reported by Shi et al. [49]. The nature of the variables studied or the variability of the means was assessed by biostatistics software Prism 5.0 (Graph-Pad™ Inc., San Diego, CA, USA). Data are expressed as the mean ± SEM. Data from 17-β-estradiol levels, body fat and uterine weight as well as CPP and IHR were analyzed by one-way analysis of variance (ANOVA), with physical training considered as the main factor. The ANG buy BIBF 1120 II-induced vasoconstriction was analyzed using a two-way ANOVA, with physical training and the concentrations of ANG II employed next were considered the main factors. In both cases, the differences among groups were determined by Tukey’s

post hoc test for multiple comparisons. Statistical significance was set at p < 0.05. Plasma 17β-estradiol concentration and the uterus weight (UW) were used to determine the estrogenic status. As expected, there was a significant decrease in both of these parameters in OVX animals (p < 0.05) when compared with the SS and STS groups ( Fig. 1A and B). Table 1 shows the body weight (BW) at the beginning and end of the study. There were no differences in BW among all groups before the experimental period; however, all groups, except for the STS group, had an increased BW after the experimental period (p < 0.05). Fig. 2 shows the fat pad values. The RET and PME fat pad weight in STO group was significantly less than the SO group (p < 0.05). In the RET and PME fat pad weight in STS group was significantly less than the SS group (p < 0.05), demonstrating the efficacy of ST in reducing adiposity. Moreover, the SO group showed an increased RET and PME fat pad weight compared with the SS group (p < 0.05). The PR values did not change among the groups tested. The inguinal fat pad was increased in both ovariectomized groups compared with the SS group (p < 0.05); however, the inguinal fat pad in the STO group was also increased compared with the STS group (p < 0.05).