Objective: To investigate the role of heat shock transcription factor 1 (HSF1) in inflammatory response induced by high mobility group protein B1 (HMGB1). Methods: The TNF-α contents in the supernatants of RAW264.7 cells after exposure to different concentrations of HMGB1 for different times were determined by ELISA assay. The NF-κB nuclear translocation and HSF-1 expression in RAW264.7 cells after HMGB1 treatment were examined by immunofluorescence and Western blot, respectively. The change in TNF-α expression induced by HMGB1 in RAW264.7 cells after interference of HSF-1 expression was observed. Results: RAW264.7 cells presented two peaks of TNF-α release at 4 and 12 h respectively after HMGB1 treatment, and the TNF-α release level was increased with the elevation of HMGB1 concentration. The NF-κB nuclear translocation was significantly enhanced and HSF-1 expression was significantly increased in RAW264.7 cells after HMGB1 treatment (both P<0.05). The TNF-α expression induced by HMGB1 in RAW264.7 cells after interference of HSF-1 expression was significantly increased compared with RAW264.7 cells without HSF-1 interference (P<0.05). Conclusion: HMGB1 can induce inflammatory response and HSF-1 expression in RAW264.7 cells, but HSF-1 may play a negative feedback inhibitory role in the HMGB1-induced inflammation.