A murine model that temporarily interrupts pulmonary arterial flow while maintaining alveolar ventilation was employed as perviously reported17 (link). In short, mice were anesthetized with avertin (intraperitoneal), intubated, mechanically ventilated and buprenorphine was administered (intraperitoneal) for analgesia. A left thoracotomy was performed and the left pulmonary artery isolated from the left bronchus. A slip knot (8-0 prolene) placed around the pulmonary artery was used to occlude pulmonary arterial flow for time periods ranging from 30 minutes to 2 h (ischemia time). One end of the suture utilized to make the slip knot was externalized to allow the tie to be released at the end of the ischemic period. The left lung was reinflated using positive pressure, the thoracotomy closed, the mouse extubated, and allowed to recover from anesthesia. The period of reperfusion lasted 1–3 h, after which the lungs were harvested and divided for RNA preparation and hematoxylin and eosin (H&E) staining.
For the sham procedure, mice received a thoracotomy but the left pulmonary artery was not isolated. The left lung was reinflated using positive pressure, after which the thoracotomy was closed and the mouse extubated. The mouse was sacrificed after a cumulative period of 30 minutes (analogous to the ischemic period) and additional 1–3 h (analogous to the reperfusion period).
For the sham procedure, mice received a thoracotomy but the left pulmonary artery was not isolated. The left lung was reinflated using positive pressure, after which the thoracotomy was closed and the mouse extubated. The mouse was sacrificed after a cumulative period of 30 minutes (analogous to the ischemic period) and additional 1–3 h (analogous to the reperfusion period).
