〔Abstract〕 To explore the mechanism of hesperidin on hindlimb motor function in mice with spinal cord injury (SCI). Methods Oxygen glucose deprivation (OGD) was used to induce SCI in PC12 neuronal cells. CCK-8 assay determined the optimal hesperidin concentration (0, 10, 20, 30, 40, 50, 60 mmol/L) for subsequent experiments after 24h treatment. PC12 cells were divided into Control group, OGD group and Hesperidin (30 mmol/L)+OGD groups. Hoechst staining assessed apoptosis; Western blot (WB), qPCR and immunofluorescence (IF) detected the expression of iNOS, CD206 and TNF-α . Flow cytometry measured apoptosis rates, and WB examined the impact of hesperidin on PI3K, p-PI3K, AKT, p-AKT, and NF-κB expression. Forty-five healthy male C57BL/6 mice were randomly divided into Sham group, Sci group and Hesperidin groups. Sham and Sci groups received 0.1 mL PBS, while the Hesperidin group received 0.1 mL hesperidin. After modeling, iNOS, CD206, and TNF-α expression levels, spinal cord cavitation, and hindlimb motor function were evaluated. Results Hesperidin at 10–30 mmol/L for 24h did not significantly affect PC12 cell activity. Hoechst staining showed reduced apoptosis in PC12 cells after 24h of 30 mmol/L hesperidin treatment. Compared to the OGD group, Hesperidin+OGD group had decreased mRNA and protein expression of inflammatory factors (iNOS and TNF-α) and increased expression of the anti-inflammatory factor (CD206). Hesperidin treatment inhibited PI3K/AKT/NF-κB pathway activation, with similar results in mouse experiments. Compared to Sci group, Hesperidin group had reduced spinal cord cavitation area and improved hindlimb motor function. Conclusion Hesperidin may improve hindlimb motor function in mice with SCI by downregulating the phosphorylation level of the PI3K/AKT/NF-κB pathway, inhibiting inflammatory factor release, promoting anti-inflammatory factor release, regulating the inflammatory microenvironment after SCI, and suppressing the acute inflammatory response.