This study was designed to determine the effect of all-trans retinoic acid (RA) on the development of cardiac remodeling in a pressure overload rat model. Male Sprague-Dawley rats were subjected to sham operation and the aortic constriction procedure. A subgroup of sham control and aortic constricted rats were treated with RA for 5 mos after surgery. Pressure overloaded rats showed significantly increased interstitial and perivascular fibrosis, HW/BW ratio (heart weight/body weight) and gene expression of atrial natriuretic peptide and brain natriuretic peptide. Echocardiographic analysis showed that pressure overload induced systolic and diastolic dysfunction, as evidenced by decreased fractional shortening, ejection fraction, stroke volume and increased E/Ea ratio and isovolumic relaxation time. RA treatment prevented the above changes in cardiac structure, function and hypertrophic gene expression in pressure overloaded rats. RA restored the ratio of Bcl-2/Bax, inhibited cleavage of caspase-3 and 9, and prevented the decreases in the levels of SOD-1 and SOD-2. Pressure overload induced phosphorylation of ERK1/2, JNK and p38 was inhibited by RA, via upregulation of MKP-1 (mitogen-activated protein kinase phosphatase) and MKP-2. The pressure overload induced production of angiotensin II was inhibited by RA, via upregulation of expression of ACE2 and through inhibition of the expression of cardiac and renal renin, angiotensinogen, angiotensin converting enzyme and angiotensin type 1 receptor. Similar results were observed in cultured neonatal cardiomyocytes, in response to static stretch. These results demonstrate that RA has a significant inhibitory effect on pressure overload-induced cardiac remodeling, through inhibition of the expression of RAS components.