Objective: To investigate the inhibitory effect of heat shock transcription factor 1 (HSF-1) on the inflammatory response induced by high-mobility group box-1 protein (HMGB1) and the mechanism. Methods: RAW264.7 cells were transfected with plasmid bearing HSF-1 gene (HSF-1 overexpression group) or empty plasmid (negative control group) respectively, using the untreated RAW264.7 cells as blank control group, and then the HSF-1 protein expression in each group of cells was determined by Western blot analysis. The above three groups of cells were stimulated with HMGB1 (1 μg/mL), and then in each groups of cells, the TNF-α levels were measured by ELISA assay, the proteins associated with mitogen-activated protein kinases (MAPK) and NF-κB pathways were detected by Western blot analysis, and NF-κB DNA binding activity was analyzed by electrophoretic mobility shift assay (EMSA), respectively. Results: Western blot analysis showed that the expression level of HSF-1 protein in HSF-1 overexpression group was significantly increased compared with negative control group or blank control group (both P<0.05), which had no significant difference between the two latter groups (P>0.05); after exposure to HMGB1 for 4 h, the TNF-α level in HSF-1 overexpression group was significantly decreased compared with negative control group or blank control group (both P<0.05), which had no significant difference between the two latter groups (P>0.05); after exposure to HMGB1 for different time periods, the expressions of MAPK pathway related proteins that included p-ERK, p-JNK and p-p38 as well as NF-κB pathway related protein p-IKα-B all showed no significant difference among the groups of cells (all P>0.05); results of EMSA showed that the expression of the gray-scale for NF-κB in HSF-1 overexpression group was significantly lower than that in negative control group or blank control group (both P<0.05), which had no significant difference between the two latter groups (P>0.05). Conclusion: HSF-1 overexpression can inhibit the TNF-α expression induced by HMGB1, and the mechanism may be associated with the NF-κB DNA binding activity but irrelevant to the MAPK signaling pathway.