Male adult Sprague–Dawley rats, weighing 270–300 g, obtained from the Animal Center of the Fujian Medical University, were anesthetized with intraperitoneal sodium pentobarbital (50 mg/kg). After intravenous injection of heparinized (50 IU) the hearts were quickly excised and fixed on a Langendorff apparatus (Radnoti, Monrovia, CA, USA) and perfused with 37°C Krebs–Henseleit buffer (KHB) at constant pressure (75 mmHg). After equilibrated with KHB for 15 min, isolated hearts were randomly divided into five groups: 1. Sham group: hearts subjected to 195-min perfusion without I/R injury; 2. I/R group: hearts received 30- min ischemia followed by 120-min reperfusion; 3. SPC group: 2.5% sevoflurane preconditioning was gived for 15 min followed by 15-min washout before ischemia; 4. SPC+MF1 group: combined application of MF1(ImClone, Somerville, NJ, USA), rat anti-VEGFR-1 monoclonal antibody,10 μmol/L and 2.5% sevoflurane preconditioning for 15 min followed by 15-min washout before ischemia; 5. SPC+PIGF group: combined application of PIGF(Abcom, Cambridge, United Kingdom)10 μmol/L and 2.5% sevoflurane preconditioning for 15 min followed by 15-min washout before ischemia. To implement sevoflurane preconditioning, the KHB was pre-equilibrated with sevoflurane under a separate reservoir with a Vapor 2000 (Dräger Medical AG & Co., Lübeck, Germany) using an air bubbler.