The invention discloses a preparation method of a rat myocardial ischemia reperfusion model. The preparation method comprises the following steps of: S1, preparing surgical instruments required by anexperiment as follows: 10% water, a 1ml injector, iodophor, a cotton swab, an electric clipper, a surgical plate, four rubber bands, scissors, two large ophthalmic bent forceps, a needle holder, hemostatic forceps, a 6-0 needle suture line, a 3-0 suture line, a suture needle, an electronic scale and a respirator; and S2, selecting a rat which is in a relatively good state, is male and has the weight of 220-250 g, weighing the rat, anesthetizing the rat, lying the rat on the back, fixing the rat on the surgical plate by using the rubber bands, shaving the left side of the sternum, wiping the iodophor on the shaved part for disinfection, longitudinally cutting the skin on the left side of the sternum by 4 cm, bluntly separating subcutaneous muscles, and exposing the pleura between the separated third and fourth ribs. Compared with an existing preparation method of a mouse myocardial ischemia reperfusion model, the preparation method of the rat myocardial ischemia reperfusion model has the advantages that the control is simple, the survival time of the rat is long, and the perfusion effect is good.