Objective To investigate the effect of hydrogen-rich saline on apoptosis in hippocampal neurons induced by cerebral ischemia-reperfusion in rats and PI3K/Akt/FoxO1 signaling pathway. Methods The rat model of focal cerebral ischemia-reperfusion was established by thread-occlusion of the middle cerebral artery in rats. SD rats were randomly divided into sham operation group (Sham group), ischemia-reperfusion group (I/R group) and hydrogen-rich saline treatment group (HRS group), 10 rats in each group. At 24 h after reperfusion, the serum levels of IL-6, TNF-a and IL-1β were detected by ELISA. Histological changes of the hippocampus were observed by pathology using HE staining. Apoptosis in brain tissues was observed by TUNEL staining. The expression changes of p-PI3K, Akt, caspase-3 and FoxO1 proteins were detected by Western blot assay. Results Compared with the sham group, pyramidal cells were arranged loosely in the I/R group and a large number of pyramidal cells were necrotized, and the amount of apoptotic hippocampal cells was increased. The levels of IL-1β, IL-6 and TNF-α were significantly increased (P< 0.05), as well as the expression of p-PI3K, Akt and caspase-3 in the brain tissue. However, the expression of FoxO1 protein was decreased. There were significant differences between the two groups (P< 0.05). Compared with the I/R group, the inflammatory factors were significantly decreased in the HRS group. The expressions of p-PI3K, Akt and caspase-3 were also significantly decreased, while the expression of FoxO1 protein was increased (P< 0.05). Conclusions Hydrogen-rich saline can reduce brain injury caused by ischemia-reperfusion, and its mechanism may be related to PI3K/Akt/FoxO1 signaling pathway.