Inflammation has been known to be involved in the pathogenesis of mastitis. And anti-inflammatory agent is proposed to be a possible efficient therapeutic strategy for mastitis. Corynoline, a bioactive compound extracted from Corydalis bungeana Turcz., has been reported to have anti-inflammatory effect. However, whether corynoline has protective effect against mastitis remains unclear. The aim of this study was to evaluate the protective effect of corynoline on LPS-induced mastitis in mice. Inflammatory cytokine production was measured by ELISA. The proteins of signaling pathways were detected by western blot analysis. The results showed that treatment of corynoline at the doses of 15, 30, and 60 mg/kg significantly attenuated LPS-induced pathological damage of mammary tissues. Corynoline also ameliorated LPS-induced MPO activity, MDA content, and inflammatory cytokine TNF-α and IL-1β production in mammary tissues. LPS-induced NF-κB activation was inhibited by corynoline. Furthermore, our results showed corynoline significantly increased the expression of Nrf2 and the phosphorylation levels of AKT and GSK3β. In conclusion, our results indicated that corynoline protected against LPS-induced mastitis through regulating AKT/GSK3β/Nrf2 signaling pathway, which subsequently led to the inhibition of NF-κB and inflammatory response. © 2021 Wiley Periodicals LLC.