Abstract: In this study, normal isolated rat heart model was adopted to investigate the inotropic and electrocardiac effect of AF-HF001 with Lanfendorff technique on the left ventricular systolic pressure(LVSP), left ventricular end diastolic pressure(LVEDP), peak rate of rise of left ventricular pressure(+dp/dt_max)and peak rate of fall of left ventricular pressure(-dp/dt_max). The results indicated that AF-HF001 could significantly enhance LVSP, LVEDP and ±dp/dt_max in a dose-dependent manner and exerted a positive inotropic effect on normal isolated rat hearts. This study also analyzed the electrocardiac effect of AF-HF001 with the electrodes located on tip of heart, ventricular free wall and aortic root. The results exhibited that AF-HF001 did not change PR, QRS, QT or HR at various concentrations, which means that AF-HF001 could safely increase cardiac contractility. This research lays a good foundation for the further study on AF-HF001 and research on structure-activity relationship of its analogs.