Animal model of Ischemia/Reperfusion

Ischemia was induced by coronary artery ligation. Twelve-week- old mice were intraperitoneally anesthetized with ketamine/xylazine (100 and 10 mg/kg, respectively), intubated with a 1-mm steel tube, and ventilated (2ml, 80 strokes/min). The thorax was opened between the second and the third ribs and widened with the aid of a mouse retractor. The pericardium was opened and the left anterior descending (LAD) coronary artery was occluded for 30 min close to its bifurcation with a 6–0 silk suture. Reperfusion was performed by releasing suture, 30 minutes after LAD occlusion. The chest was closed, negative pressure was reestablished, and the skin was sutured. Control mice (sham) were included in the assays, in which the same procedure was performed except for LAD occlusion.