Integrating Network Pharmacology and Experimental Validation: Mechanistic Study of the Anti-Porphyromonas gingivalis and Anti-Inflammatory Effects of Berberis hemsleyana Ahrendt Extract

Anti-<i>Porphyromonas gingivalis</i> mechanisms of <i>Berberis hemsleyana</i> bark extract remain to be elucidated, and the anti-inflammatory activity of its n-butanol fraction (BNB) in RAW264.7 cells-mediated through suppression of the NF-κB pathway-require further validation. The minimum inhibitory concentration (MIC) and minimum bactericidal concentration (MBC) of the crude extract from <i>B. hemsleyana</i> were determined against <i>Candida albicans</i>, <i>Escherichia coli</i>, <i>Porphyromonas gingivalis</i>, <i>Staphylococcus aureus</i> and <i>Streptococcus mutans</i>. Scanning electron microscopy (SEM) and bacterial protein leakage assays were used to evaluate its antibacterial activity against <i>P. gingivalis</i>. High-performance liquid chromatography-mass spectrometry (LC-MS) was applied to analyze the ethanol extract of <i>B. hemsleyana</i> bark, leading to the screening of 47 compounds. The antibacterial mechanisms of the compounds were predicted through Network Pharmacology analysis and Molecular docking. Anti-inflammatory activity mediated via the NF-κB pathway was verified using an LPS-induced RAW264.7 cell inflammatory model. Specifically, the BNB showed a significant antibacterial effect on <i>P. gingivalis</i>. Meanwhile, it was confirmed that this fraction damaged the bacterial cell membrane structure, leading to the leakage of intracellular proteins in bacteria and thus impairing their infectivity. Network pharmacology analysis and molecular docking results indicated that <i>B. hemsleyana</i> bark's biologically active compounds (Calenduloside E, Limonin, Acanthoside B, Dihydroberberine) antibacterial activity by regulating cytokines and cell apoptosis, thereby coordinating the body's microbial homeostasis and inflammation. Additionally, BNB significantly reduced the secretion of the inflammatory cytokines IL-1β, TNF-α and IL-6 in vitro via the NF-κB pathway. The crude extract from the bark of <i>B. hemsleyana</i> has antibacterial and Anti-inflammatory activity. The n-butanol fraction showed a significant antibacterial effect on <i>P. gingivalis</i>.