Abstract As a flavonoid compound, astragalin (AST) is found in a variety of medicinal plants. In clinical studies, AST has anti-inflammatory and analgesia effects on rheumatoid arthritis, bronchopneumonia diseases and so on, but its specific role and mechanism is still not clear. This study aimed to investigate the effect and molecular mechanism of AST on inflammatory pain and pain-related emotions in complete Freund’s adjuvant (CFA) mice. In this study, we observed that AST significantly alleviated CFA-induced inflammatory pain and associated emotional disturbances in mice. The mechanism may be related to down-regulating mGluR5-mediated PKCλ-ERK1/2-FOXO6 signaling pathway in CFA mice. Treatment with the mGluR5-specific inhibitor MTEP resulted in the downregulation of proteins associated with the PKCλ-ERK1/2-FOXO6 pathway, similar to the effects observed with AST administration. These results suggested that AST might play a crucial role in the management of inflammatory pain and related emotions, shedding light on its underlying mechanism for treating such conditions.