Renal fibrosis is a common pathological feature of the progression of chronic kidney disease. Although valproic acid (VPA) has been recently shown to induce autophagy, the effect of VPA-induced autophagy on renal fibrosis remains unknown. We, therefore, investigated whether VPA-induced autophagy suppresses renal fibrosis in a mouse model of unilateral ureteral obstruction (UUO).Male C57BL/6 mice were divided into five groups (n = 8 per group): (1) sham group; (2) vehicle group; (3) VPA-treated group; (4) 3-methyladenine (3-MA; autophagy inhibitor)-treated group; and (5) VPA plus 3-MA-treated group. Mice underwent UUO and the kidneys were studied after 5 days. We also investigated the effect of VPA-induced autophagy on alpha-smooth muscle actin (alpha-SMA) in transforming growth factor (TGF)-beta 1-stimulated rat kidney fibroblasts and epithelial cells.VPA attenuated renal fibrosis and induced autophagy in UUO mice, while 3-MA increased renal fibrosis and suppressed autophagy. In addition, the anti-fibrotic effect of VPA was diminished by 3-MA in UUO mice. In rat kidney fibroblasts and epithelial cells, VPA suppressed TGF-beta 1-stimulated alpha-SMA expression and induced autophagy. In contrast, 3-MA enhanced alpha-SMA expression while inhibiting autophagy. Furthermore, the combined use of VPA and 3-MA treatments increased the expression of alpha-SMA compared with VPA treatment alone in TGF-beta 1-stimulated rat kidney fibroblasts and epithelial cells, which was accompanied by the inhibition of autophagy.These findings suggest that VPA may be a candidate drug for the treatment of renal fibrosis through the induction of autophagy.