Objective: To investigate the effect of resveratrol on TNF-α promoter region in human monocyte THP-1 cells tolerated to bacterial lipopolysaccharide (LPS). Methods: The LPS-tolerated THP-1 cells were induced, next the untreated THP-1 cells (control group), LPS-tolerated THP-1 cells (tolerance group) and resveratrol treated LPS-tolerated THP-1 cells (tolerance plus resveratrol group) were stimulated with LPS respectively, and then, the TNF-α mRNA expression and the bindings of transcription factors to the promoter region of TNF-α were determined. Results: After LPS stimulation, the TNF-α mRNA level was increased rapidly in control group, and increased slowly in tolerance group, but were slightly decreased in tolerance plus resveratrol group, which was significantly lower than that in either former group (both P<0.05). Results of chromatin immunoprecipitation (ChIP) showed that before LPS simulation, the levels of p65 along with acetylated p65 (ace-p65), RelB and G9a binding to the promoter region of TNF-α in both tolerance group and tolerance plus resveratrol group were significantly higher than those in control group (all P<0.05), while the binding level of p50 had no significant difference among the three groups (P>0.05); after LPS simulation, the levels of p65 and ace-p65 binding to the promoter region of TNF-α were significantly increased in control group, but significantly decreased in tolerance plus resveratrol group, and level of G9a binding to the promoter region of TNF-α was decreased in control group (all P<0.05), and all other factors had no significant change compared with those before LPS stimulation (all P>0.05). Conclusion: Resveratrol can inhibit TNF-α mRNA expression in LPS-tolered THP-1 cells, which may be partially associated with its inhibiting the bindings of p65/ace-p65 to TNF-α promoter. So, Resveratrol may potentially be used in supplementary treatment of sepsis.