The aim of the present study was to investigate the effect of ligustilide (LIG) on angiotensin II (Ang II)-induced hypertrophy in neonatal rat myocardial cells and the expression levels of p53, Bcl-2 and Bax. Myocardial cells were isolated and purified from the ventricles of neonate Sprague-Dawley rats (age, 1–3 days) using a differential adhesion method. The cells were then were stimulated by Ang II and LIG for 1–3 days, following which the cell surface area, intracellular protein concentration, rate of apoptosis and the expression levels of p53, Bcl-2 and Bax were determined. Following stimulation with Ang II, the cell surface area of the neonatal rat myocardial cells increased significantly and the cell morphology was distorted. LIG was shown to significantly suppress the Ang II-induced hypertrophy of neonatal rat myocardial cells in a dose-dependent manner. In addition, administration of LIG restored the expression levels of p53, Bcl-2 and Bax. Therefore, LIG can prevent the hypertrophy of cardiomyocytes induced by Ang II, which may be associated with the inhibitory effect that LIG exhibits on cardiomyocyte apoptosis.