The expression levels of microrna (mir)-340-5p are reportedly decreased in the peripheral blood during acute ischemic stroke; however, the direct effect and mechanism of action of mir-340-5p in ischemic stroke remains largely unknown. The present study aimed to investigate the effects of mir-340-5p, and its mechanism of action, on Pc12 cells following oxygen-glucose deprivation/reperfusion (oGd/r) induction. oGd/r-induced Pc12 cells served as the cellular model and subsequently, mrna expression levels of mir-340-5p and neuronal differentiation 4 (neurod4) were analyzed using reverse transcription-quantitative Pcr. Tumor necrosis factor-α, interleukin (il)-1β and il-6 expression levels were detected using ELISA kits, and flow cytometry was used to determine the rate of cellular apoptosis. in addition, a nitric oxide (no) synthase activity assay kit was used to detect no levels and a nadPH assay kit was used to measure nadPH levels. Western blotting was also performed to analyze protein expression levels of bax, bcl-2, cleaved caspase 3 and phosphorylated endothelial noS (enoS), and the target gene of mir-340-5p was predicted using TargetScan software and verified using a dual-luciferase reporter assay. The expression levels of mir-340-5p were decreased in Pc12 cells following oGd/r induction and Neurod4 was identified as a target gene of miR-340-5p. In addition, miR-340-5p overexpression reduced inflammation, apoptotic rate, no production and nadPH levels, in addition to increasing enoS expression in Pc12 cells following oGd/r induction. notably, the overexpression of neurod4 reversed the aforementioned effects of mir-340-5p on Pc12 cells following OGD/R induction. In conclusion, the findings of the present study suggested that mir-340-5p may protect Pc12 cells against oGd/r through targeting neurod4, which could provide important implications for the treatment of ischemia-reperfusion injury based on mir-340-5p expression levels in vivo.