In addition to ACE2 and Ang-(1-7), Mas is part of the cardioprote

In addition to ACE2 and Ang-(1-7), Mas is part of the cardioprotective axis of the RAS, therefore it is of major importance to determine whether cardiac Mas expression is modulated

at distinct physiological and pathological conditions. In our recent report, we have submitted Wistar rats and spontaneously hypertensive rats (SHRs) to a physical training protocol. Interestingly, only SHR presented an increase in left ventricular Mas expression in response to exercise training [9], indicating that cardiac Mas expression can be regulated not only according to the stimulus, physiological or pathological, but also according to the state of the animal, i.e. healthy or diseased. Therefore, the aim of this study was to evaluate the responsiveness of Mas expression in rat hearts submitted to pathological challenges such as myocardial hypertrophy, MG-132 order infarction and hypertension, and under a physiological condition (physical exercise training). Three month-old male Wistar and Sprague-Dawley (SD) rats were used in this study. The animals were provided by the animal facility of the Biological Sciences Institute (CEBIO, Federal University

of Minas Gerais) and housed in a temperature (22–24 °C) and humidity-controlled room maintained on a 12:12-h light–dark schedule with free access to food and water. All animal procedures were performed in accordance with guidelines for the humane use of laboratory animals at our Institute and were approved by local authorities. Selleck MDV3100 The exercise training was performed in swimming pools with controlled temperature (31 ± 1 °C) for 40–60 min per day, 5 days per week over 10 weeks.

After the first week of adaptation in the swimming pools, Wistar rats (3 month-old, n = 4–6) were submitted to a progressive load test, which consisted of an increasing workload corresponding to 2% of body weight added every 3 min until exhaustion. This test was repeated at the end of the physical training protocol. Exercise intensity of the endurance training was set at 50–80% (second and third weeks: 40–60 min at 50%; fourth week: 40 min at 60%; fifth week: 40 min at 70%; and sixth to tenth weeks: 40–60 min at 80%) of the maximal weight obtained in the progressive test. The maximal weight carried by the Celecoxib animal in the progressive load test was converted to percentage of the animal body weight. Thus, every week the rats were weighed, and using the previously calculated percentage value, a new maximal load was obtained and the 50–80% workload was determined. With this procedure, we eliminated the need for performing the progressive load test on a weekly basis [1]. At the end of the training, the rats were killed by decapitation and the hearts were immediately removed. Left ventricular wet weights were recorded, normalized for body weight and then expressed as cardiac mass index (mg/g). The left ventricles were used for histology and western blot analysis.

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