Myocardial fibrosis is the fundamental remodeling process in myocardial ischemia (MI) and also the major contributor of heart failure and death. Tanshinol (Danshensu in Chinese, DSS), a major ingredient of salvia mitiorrhiza Bunge (Lamiaceae) root, exerted significant cardio protection effects. In this study, we aimed to identify the action target and then uncover the mechanism of DSS alleviating myocardial fibrosis. The pharmacological activities of DSS protecting ischemic cardiac was assessed and the myocardial proteomics was carried out. To identify the target of DSS, a cellular thermal shift assay combined with LC-MS identification was conducted. Surface plasmon resonance assay, molecular dynamics simulation and pharmacological and molecular biology approaches were adopted to explore the action mechanisms of DSS. Our results revealed that DSS effectively alleviated MI-induced left ventricle dysfunctions and the increasements of circulating myocardial markers. Besides, DSS significantly reversed the proteomic profile related to myocardial fibrotic processes and the ERK2 was identified as a crucial cellular target of DSS. DSS abated the temperature-dependent denaturation of ERK2 in a dose-dependent manner and the K_D value of DSS and ERK2 was 60.19 μM. After Ang II stimulation, DSS suppressed the phosphorylation of Thr188 rather than the classic residues in TEY motif. DSS interfered the ERK2 homo-dimerization and then blocked the intermolecular autophosphorylation at Thr188 site. Thereout, DSS inhibited the nuclear translocation of ERK2 and the expression of downstream fibrotic biomolecules. Collectively, our results demonstrated that DSS targeted ERK2 and suppressed the intermolecular autophosphorylation at Thr188 residue, thus protecting ischemic myocardia from fibrosis remodeling.