Using gene-targeted TRPV1 (transient receptor potential vanilloid type 1 channels)-null mutant (TRPV1^-/-) or wild-type (WT) mice, we test the hypothesis that TRPV1 contributes to protease-activated receptors 2 (PAR2)-mediated cardiac protection via increasing the release of calcitonin gene-related peptide (CGRP) and/or substance P (SP). Immunofluorescence labeling showed that TRPV1 coexpressed with PAR2, PKC, or PKAc in WT but not TRPV1^-/- hearts. Hearts were Langendorffly perfused with the selective PAR2 agonist, SLIGRL, followed by global ischemia and reperfusion (I/R). The recovery rate of coronary flow (% CF), the maximum rate of left ventricular pressure development (dP/dt), left ventricular end-diastolic pressure (LVEDP), and left ventricular developed pressure (LVDP) were evaluated after I/R. SLIGRL improved the recovery of hemodynamic parameters, decreased lactate dehydrogenase (LDH) release, and reduced the infarct size in both WT and TRPV1^-/- hearts (p<0.05). The protection of SLIGRL was surpassed for WT compared to TRPV1^-/- hearts (p<0.05). CGRP8-37, a selective CGRP receptor antagonist; RP67580, a selective neurokinin-1 (NK1) receptor antagonist; PKC V1-2, a selective PKC inhibitor; or H-89, a selective PKA inhibitor, abolished SLIGRL protection in WT but not TRPV1^-/- hearts. SLIGRL increased the release of CGRP and SP in WT but not TRPV1^-/- hearts (p<0.05), which were prevented by PKC V1-2 and H-89. Our data show that PAR2 activation improves cardiac recovery after I/R injury in WT and TRPV1^-/- hearts with a greater effect in the former, suggesting that PAR2-mediated protection is TRPV1-dependent and -independent, and that dysfunctional TRPV1 impairs PAR2 action. PAR2 activation of the PKC or PKA pathway stimulates or sensitizes TRPV1 in WT hearts, leading to the release of CGRP and SP that contribute at least in part to PAR2-induced cardiac protection against I/R injury.