Myocardial ischemia/reperfusion (MI/R) injury is a complex pathological process, which causes increased mortality and disability even after treatment of coronary heart disease. Currently, there are few clinical treatments for MI/R injury. Many studies have shown that MI/R injury can cause Ca²⁺ overload that can further aggravate MI/R injury. One of Ca²⁺ permeable channels, transient receptor potential vanilloid 2 (TRPV2) has been suggested as a candidate for Ca²⁺ entry pathways and a potential therapeutic target for MI/R. Here, we examined the involvement of TRPV2 in the pathology of MI/R of mice and assessed whether the experimental blockade of surface accumulation of TRPV2 ameliorates MI/R injury. On reperfusion following global ischemia, wild type mouse hearts exhibited surface expression of TRPV2, low left developed pressure, and increased release of creatine kinase. We observed that overexpression of the amino-terminal (NT) domain of TRPV2 could block the plasma membrane accumulation and influx of Ca²⁺ via TRPV2 in HEK293 cells. Transgenic expression of the NT domain under the control of the alpha-myosin heavy chain promoter in mice caused an effective removal of sarcolemmal TRPV2 that was activated in MI/R and significantly improved the cardiac function and attenuated creatine kinase release in MI/R. These results suggest that cardiac TRPV2 activation likely contribute to MI/R injury and, thus, that TRPV2 may be a novel therapeutic target for reducing MI/R injury.