Abstract: The effects of polymyxin B (PXB) on action potential of the mice right ventricular myocardial cell were studied by means of intracellular capillary glass electrode. The action potential amplitude (APA) and maximal depolarization rate of phase O (V_max) of action potential were lowered after administering PXB for 5 min. APA was lowered from 104.0±4.2 mV to 100.7±5.1mV, decreasing 3.2% (P<0.01); V_max was lowered from 691.5±114.1 v/s to 515.15±71.5 v/s, decreasing 25.5% (P<0.001). Practically, the resting potential remained unchanged. With the continued administration of PXB, the functional refractory period was significantly prolonged. APA₉₀ was temporarily shortened and then prolonged. The results suggest that PXB inactivate or block the fast sodium channel in myocardial cell membrane, which may relate to the antiarrhythmic effect in animal experiment.