<p>Schisandrin is one of the main components isolated from Schisandra fruit (<i>Schisandra chinesnesis</i>) and has been prescribed for hepatitis treatment. Recent studies have demonstrated that Schisandrin shows anti-inflammation, anti-oxidative and anti-allergic effects in vivo and in vitro models. However, the fundamental mechanism of anti-inflammation by Schisandras has not been fully evaluated. Here, we aimed to study the anti-inflammatory role through the modulation of High mobility group box 1 (HMGB1)/Toll-like receptor 4 (TLR4) signaling pathway by Schisandrin. Schisandrin inhibited HMGB1 expression and translocation, reduced HMGB1/TLR4 binding and lipid raft recruitment, and suppressed the activation of Nuclear Factor-kappa B (NF-κB) and Mitogen-Activated Protein Kinase (MAPK) pathways. These effects led to a concentration-dependent reduction in LPS-induced inflammation, suggesting that Schisandrin attenuates inflammation by modulating HMGB1/TLR4-mediated signaling. These findings provide novel insights into the molecular basis of Schisandrin's anti-inflammatory activity and highlight its potential as a therapeutic agent for HMGB1/TLR4-associated inflammatory diseases.</p>