OBJECTIVE To study the diastolic functions using color Doppler M-mode (CDMM) for noninvasive analysis of the intraventricular pressure difference (IVPD) in diabetic rats. METHODS Two equal groups of rats were included: control and streptozotocin-induced DM (n = 15). The cardiac functions were examined monthly using conventional echocardiography and CDMM with a specific MATLAB software. Echocardiography was performed under 2% isoflurane mask inhalation. Five months thereafter, all rats were killed for macroscopic and microscopic examinations of the cardiac fibrosis. RESULTS DM rats showed higher systolic blood pressure and diastolic dysfunction, i.e., decreases in several parameters such as E, E/A, TDIs, and IVPDs, compared to the controls. Moreover, obvious cardiac fibrosis was seen in perivascular and interstitial tissues, but there were no notable differences in terms of gross lesions. CONCLUSIONS Because of the noninvasive nature of CDMM, IVPD and other conventional echocardiographic parameters can be used as reliable indicators generally for evaluating cardiac function and particularly the change in intraventricular pressure.