Background: High Mobility Group Box 1 protein (HMGB1) is a chromatin binding nucleus protein and has proinflammatory cytokine potential when released by damaged or necrosis cells during Inflammatory bowel dieases(IBD). This study is aimed to explore the association between ISQ and HMGB1 release in human intestinal cell. Methods: The protein expression of COX-2, fracnation of NF-κB and HMGB1, concentration of culture medium were analyzed by Western blot. Translocation of NF-κ B and HMGB1 were assessed by Fluorescence staining. Co-Immunoprecipitation assay for demonstrated acetyled HMGB1 in medium. HMGB1 and COX-2 mRNA level was analyzed by RT-PCR. Results: ISQ reduce TNF-a induced release of HMGB1 in extracellular and inhibit nucleus/cytosol translocation. ISQ also regulates NF-κB p65 translocation and inhibit the COX-2 expression which is the downstream of the NF-κB. Moreover ISQ suppressed the release of HMGB1 through reduction of the mRNA level and inhibit HMGB1 acetylation. Conclusions: In this study, all data evidence that released HMGB1 is a proinflammatory cytokine and leads to signaling cascades in inflammatory responses in human intestinal cell. These findings highlight the potential of ISQ for clinical applications in the treatment of intestinal inflammation including IBD.