Renal ischemia-reperfusion (I/R), an important cause of acute kidney injury, is unavoidable during various types of operations, including renal transplantation, surgical revascularization of the renal artery, partial nephrectomy, and treatment of suprarenal aortic aneurysms. Exacerbation of I/R injury is mediated by reactive oxygen species (ROS). A recent study has shown that hydrogen has antioxidant properties. In this study, we tested the hypothesis that a hydrogen-rich saline solution (HRSS) attenuates renal I/R injury in a rodent model. Rats were treated with an intravenous injection of HRSS or control saline solution followed by renal I/R. After 24 h of treatment, we performed a histological examination and transmission electron microscopy, and measured serum levels of 8-OHdG. Histological analysis revealed a marked reduction of interstitial congestion, edema, inflammation, and hemorrhage in renal tissue harvested 24 h after HRSS treatment compared to saline administration. Renal I/R injury, which led to altered mitochondrial morphology, was also inhibited by HRSS. Furthermore, serum 8-OHdG levels were significantly lower in rats treated with HRSS and subjected to renal I/R. These protective effects were likely due to the antioxidant properties of HRSS. These results suggest that HRSS is a potential therapeutic candidate for treating various I/R diseases.