Purpose: To investigate the utility of diffusion kurtosis imaging (DKI) MRI for evaluation of renal fibrosis in rats with unilateral ureteral obstruction (UUO). Methods: Twenty-five rats had UUO, and ten rats were subjected to sham operation as control. DKI was performed on a 3.0 T MRI scanner on days 1, 3, 5, and 7 after ligation. All rats then underwent F-18-FDG dynamic PET to evaluate unilateral renal function, followed by histological analysis to examine a-smooth muscle actin (alpha-SMA) expression. DKI metrics were assessed among the time points and between two sides, and compared with maximum standardized uptake value (SUVmax), serum levels of creatinine and urea, and fibrosis marker alpha-SMA. Results: Mean kurtosis (MK) on day 7, axial kurtosis (Ka) on days 3 and 7, mean diffusivity (MD) on days 1, 3, 5, and 7, and fractional anisotropy (FA) on days 3, 5, and 7 of cortex and medulla between the UUO and contralateral sides were significantly different (all p < 0.05). Over the course of UUO progression, there were significant changes in Ka, MD and FA of medulla (all p < 0.05). FA of medulla was positively correlated with SUVmax (r = 0.641, p < 0.001), and MD of cortex was negatively correlated with urea (r = - 0.534, p = 0.001). MD of cortex was negatively correlated with alpha-SMA on UUO sides (r = - 0.710, p < 0.001). Conclusions: DKI shows the potential for noninvasive assessment of renal fibrosis and unilateral renal function induced by UUO.