Background and Purpose: Ischemia reperfusion (I/R) is the main cause of acute ischemic stroke injury. However, existing treatments for I/R injury are relatively poor, and relevant drugs need to be further explored. Experimental Approach: In this study, the neuroprotective function and related mechanism of OL-FS13 were studied through rat I/R model and PC12 cell model of oxygen glucose deprivation/reoxygenation model (OGD/R), combined with cell viability experiment, tissue H&E and Nissl staining experiment and Western blot experiment. Key Results: In this study, we identified a novel neuroprotective peptide (OL-FS13, amino acid sequence: FSLLLTWWRRRVC) from the odorous frog species Odorrana livida using a constructed cDNA library. OL-FS13 significantly alleviated brain injury from cerebral I/R in rats by reducing infarct volume in the brain, improving behavioral and histological abnormalities, and rescuing PC12 cell from damage caused by oxygen glucose deprivation/reoxygenation (OGD/R). Mechanistically, OL-FS13 increased the level of antioxidative enzymes to resist oxidative stress induced by I/R and OGD/R by activating the Nrf2/HO-1 pathway and alleviated endoplasmic reticulum (ER) stress induced by OGD/R by inhibiting the IRE1α/TRAF2/JNK pathway. Conclusion and Implications: Collectively, the novel peptide exerted significant anti-stroke activities by alleviating oxidative and ER stress via activation of Nrf2 signaling and suppression of the IRE1α/TRAF2/JNK pathway. Thus, this research provides a potential drug candidate for the clinical treatment of cerebral I/R.