Objective This study aims to investigate the effect of resveratrol on neuronal damage in rats with acute cerebral infarction (ACI) by regulating the high-mobility group protein B1 (HMGB1) /Toll-like receptor 4 (TLR4) / nuclear factor-κB (NF-κB) signaling pathway. Methods Rats were randomly separated into blank group, infarction group, resveratrol group, HMGB1 activator (recombinant rat HMGB1 protein, rRHMGB1) group, and resveratrol+ rRHMGB1 group, with 18 rats in each group. Except for the blank group, rats in all other groups were used to construct ACI models using the method of middle cerebral artery occlusion. Drug treatment began at 1 hour after successful modeling, once a day, for 7 days. Changes in neurological impairment score and cerebral infarction volume coefficient were detected. ELISA was applied to detect levels of tumor necrosis factor-α (TNF-α), interleukin-1β (IL-1β), and IL-6 in infarction region of brain tissue. Nissl staining was applied to detect the number of surviving neurons in infarction region of brain tissues. TUNEL staining was applied to detect neuronal apoptosis in infarction region of brain tissue. Western Blot was applied to detect Bcl2 associated X protein (Bax), TLR4, Cleaved Caspase-3, HMGB1, and p-NF-κB p65 protein in brain tissue. Results Compared with the blank group, the neurological damage score, cerebral infarction volume coefficient, levels of TNF- α, IL-1β, IL-6 in the infarction region of brain tissue, neuronal apoptosis rate, Bax, TLR4, Cleaved Caspase-3, HMGB1, p-NF- κB p65 protein increased in the infarction group, while the number of surviving neurons in the infarction region of brain tissue decreased (P<0.05). Compared with the infarction group, the neurological damage score, cerebral infarction volume coefficient, levels of TNF-α, IL-1β, IL-6 in the infarction region of brain tissue, neuronal apoptosis rate, and protein expression of Bax, TLR4, Cleaved Caspase-3, HMGB1, p-NF- κB p65 decreased in resveratrol group and resveratrol+rRHMGB1 group, the number of surviving neurons in the infarction region of brain tissue increased (P<0.05). Change of the corresponding indicators in the rRHMGB1 group was opposite to that in the resveratrol group (P<0.05). Compared with the resveratrol group, the neurological damage score, cerebral infarction volume coefficient, levels of TNF-α, IL-1β, IL-6 in the infarction region of brain tissue, neuronal apoptosis rate, the protein expression of Bax, TLR4, Cleaved Caspase-3, HMGB1, p-NF-κB p65 increased in the resveratrol+rRHMGB1 group, the number of surviving neurons in the infarction region of brain tissue decreased (P<0.05). Conclusion The mechanism by which resveratrol improves neuronal damage in ACI rats may be related to the inhibition of the HMGB1/TLR4/NF-κB signaling pathway. [ABSTRACT FROM AUTHOR]