This article examines berberine's biological effects and molecular mechanisms with an inflammatory response model induced by lipopolysaccharide (LPS) in human gingival fibroblasts (HGFs) using metabolomics. The viability of HGFs was determined using the cell counting kit-8 (CCK8). ELISA was used to measure inflammatory cytokines, including interleukin-6 (IL-6), interleukin-1 beta (IL-1 beta), and tumor necrosis factor-alpha (TNF-alpha). An investigation of western blots was conducted to investigate the related proteins of apoptosis. Low concentrations of berberine (0.1, 0.5, and 1 mu mol L-1) did not affect HGF growth, whereas high concentrations of berberine (5-25 mu mol L-1) significantly activated cell proliferation. Berberine suppressed the elevated secretion of IL-6, IL-1 beta, and TNF-alpha induced by LPS in HGF. Western blot analysis showed that 10 mu mol L-1 of berberine significantly inhibited LPS-induced apoptosis signaling pathway activation. Our results suggested that berberine could inhibit LPS-induced apoptosis and the production of proinflammatory mediators in HGFs cells. Berberine may be a potential therapeutic drug for the management of periodontitis.