Approximately 15 to 20 min after left anterior descending coronary artery ligation surgery, an osmotic minipump (model 2002, Durect, Cupertino, California) was implanted subcutaneously in the scapular region and connected to the ICV cannula using Tygon tubing (0.38-mm inside diameter; Cole Parmer, Vernon Hills, Illinois) to deliver leptin (0.62 μg/h), vehicle (saline, 0.5 μl/h), or the MC4R agonist melanotan II (MTII) (10 ng/h) for 28 consecutive days. The rates of leptin and MTII infusion were based on previous studies showing that these doses effectively decrease food intake and promote weight loss (20,28). On the 14th day of infusion, the rats were briefly anesthetized with isoflurane for osmotic minipump replacement. On day 28, the rats were euthanized and hearts were collected.

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