In 31 healthy subjects, there was a significant increase in left ventricular filling indexes (peak velocity of early mitral flow [E] and peak velocity of mitral flow during atrial contraction [A], and early distolic mitral annular velocity (E') and mitral annular velocity during atrial contraction [A']) after treadmill exercise; E/A and E'/A' ratios did not change after exercise. Values obtained in this study can serve as reference values in interpreting future diastolic stress testing to identify patients with exercise-induced symptoms caused by diastolic dysfunction.