The nuclear retinoic acid receptors (RARs) function as ligand-dependent transcriptional regulators and include three subtypes (RAR alpha, RAR beta and RAR gamma), which control the expression of specific gene subsets subsequent to ligand binding and to strictly controlled phosphorylation processes. Extracellular matrix (ECM) accumulation is the most important characteristic of renal interstitial fibrosis (RIF). This study was performed to investigate whether RARs were associated with ECM accumulation in the progression of RIF in rats. Eighty Wistar male rats were divided into a sham operation group (SHO) and a model group subjected to unilateral ureteral obstruction (GU) at random; n = 40, respectively. The RIF disease in GU group was established by left ureteral ligation. The renal tissues were collected at two weeks and four weeks after surgery. Protein expressions of RAR alpha, RAR beta, RAR gamma., transforming growth factor-beta l (TGF-beta 1), collagen-IV (Col-IV) and fibronectin (FN) were detected using immunohistochemical analysis, and mRNA expressions of RAR alpha, RAR beta, RAR gamma and TGF-beta 1 in renal tissue were detected by real time reverse transcription polymerase chain reaction. RIF index in renal interstitium was also calculated. When compared with those in SHO group, expressions of RAR alpha and RAR beta (protein and mRNA) were markedly reduced in the GU group (each p < 0.01). There was no marked difference for the expression of RAR gamma (protein and mRNA) between the SHO group and the GU group. The expressions of TGF-beta 1, Col-IV, FN and the RIF index in the GU group were markedly increased when compared with those in the SHO group (each p < 0.01). The protein expression of RAR alpha/RAR beta was negatively correlated with protein expression of TGF-beta 1, Col-IV or FN and the RIF index (all p < 0.01). In conclusion, the low expression of RAR alpha/RAR beta is associated with ECM accumulation in the progression of RIF in rats, suggesting that RAR alpha/RAR beta is a potentially therapeutic target for prevention of RIF.