The computerized non-invasive tail-cuff manometry system (MODEL MK-2000, Muromachikikai Co. Ltd, Tokyo, Japan) was used to measure the systolic and diastolic blood pressure of conscious SD rats (18 (link)). The blood pressures were measured for 15 min between 8:00 AM and 12:00 PM in rats individually restrained in a clear acrylic restrainer at an ambient temperature of 37°C. The average of five blood pressure recordings for each rat was taken as the blood pressure at that particular time. To ensure the reliability of the measurements, the rats were trained for one week to acclimatize them to the procedure.
After the measurement of blood pressure, the rats were then sacrificed under pentobarbital anesthesia (60 mg/kg). The kidneys and mesenteric arteries were removed quickly, lysed in a lysis buffer, sonicated, placed on ice for 1 h and centrifuged at 15,000 rpm for 30 min at 4°C. The supernatants were stored at −70°C until subjected to western blot analysis.
After the measurement of blood pressure, the rats were then sacrificed under pentobarbital anesthesia (60 mg/kg). The kidneys and mesenteric arteries were removed quickly, lysed in a lysis buffer, sonicated, placed on ice for 1 h and centrifuged at 15,000 rpm for 30 min at 4°C. The supernatants were stored at −70°C until subjected to western blot analysis.
