Abstract: AIM To study the activation of choline on M3-R in heart and observe the hemodynamic changes of rat and rabbit. METHODS A cardiac catheter was inserted into the left ventricular cavity via the right carotid artery, then the HR, LVSP, LVEDP, and ±dp/dt were measured using a polygraph system. RESULTS Choline was shown to decrease the hemodynamic assessments, such as HR, +dp/dt, LVSP and LVEDP. while the M3-R antagonist 4-DAMP (4-diphenylacetoxy-N-methylpiperidine-methiodide) showed little effect on these assessments. It was found to reverse the hemodynamic effects of choline. CONCLUSION M3 receptor agonist can produce negative inotropic and chronotropic effects on the heart of rat and rabbit.