Animal experiments and echocardiography assessment

All mice were maintained in a light/dark cycle of 12 hours/12 hours at a temperature of 21°C and 50% humidity. 

AAC was performed under anesthesia in 12-week-old, male, C57BL/6J mice, weighing between 25 and 30 g. First, the mice were anesthetized with intraperitoneal injection of pentobarbital (50mg/kg). Then, the mice were depilated on the chest with depilatory cream, and the epidermis was disinfected to prepare skin. Importantly, respiratory support was established through airway intubation, and the breathing machine of small animals was connected to maintain the breathing of mice. Taking the second rib as the landmark for the incision, cut the skin along the sternum to the first and third ribs, exposing the joint between the second rib and the sternum. Both layers of thoracic muscles should be cut, taking caution to avoid the vein. The second rib was then cut with a microscissors to expose the white thymus and fat, which should be pulled away with forceps. Be caution to avoid damaging pericardial sac, which was connected with thymus and left superior vena cava. The great vessels and upper part of the left atrial appendage can then be visualized. The ascending aorta was carefully bluntly dissected with the curved forceps from the pulmonary trunk (extreme caution should be taken, since bleeding could be deadly). Then the curved forceps are placed under the ascending aorta. Then, 7–0 silk is grasped by forceps and moved underneath the aorta, and a loose double knot is made. We use a 24-gauge needle for aorta constriction. (The size of the needle depends on the degree of stenosis and hypertrophy/failure desired). The needle is delivered through the loose double knot and placed directly above and parallel to the aorta. Then tie a surgical knot around the aorta and needle (this step should be done very quickly). Next, the needle was removed. The chest cavity is closed by suturing layer by layer. Sham group only placed needles without ligation. After being separated from the ventilator, the mice were placed on a body temperature keeping pad and put back into the feeding cage after awakening. The mice were given a normal diet after operation.

DWI mouse model was conducted in 12-week-old, male, C57BL/6J mice. The anesthesia procedure were the same as that of the AAC model. The skin of neck and upper chest was depilated and disinfected. A longitudinal 1 cm incision was made along the right side of trachea with microscissors. Bluntly dissect the right carotid artery with forceps, then ligate the distal end and tie a slipknot near the heart. FST guide wire (article number: FST18000-10, diameter 0.36 mm) was inserted into the carotid artery, and confirm that the tip of the guide wire enters the ascending aorta and the aortic valve at the outlet of the left ventricular outflow tract with the ultrasonic assistance of a small animal ultrasonic machine (Visual sonics Vevo 2100). Aortic valve injury was induced by scratching the leaflets with the body of the wire for 50 times and spinning the tip of the wire correctly positioned on the LV side of the valve for 100 times. Sham group was performed in the same way, but the guide wire did not enter the ventricle. Remove the guide wire and quickly ligate the right carotid artery. Suture the skin, put it on body temperature keeping pad, and put it back in the feeding cage after awakening. The mice were given a normal diet after operation.