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

## TL;DR

This study explores how an extract from Berberis hemsleyana fights bacteria and reduces inflammation, using both experiments and network analysis.

## Contribution

The paper combines network pharmacology with experimental validation to reveal the anti-bacterial and anti-inflammatory mechanisms of Berberis hemsleyana extract.

## Key findings

- The n-butanol fraction of B. hemsleyana extract significantly damages P. gingivalis cell membranes and reduces bacterial infectivity.
- Network pharmacology and molecular docking identified key compounds like Calenduloside E and Dihydroberberine that regulate inflammation and microbial balance.
- The extract reduces inflammatory cytokines IL-1β, TNF-α, and IL-6 via suppression of the NF-κB pathway in RAW264.7 cells.

## Abstract

Anti-Porphyromonas gingivalis mechanisms of Berberis hemsleyana 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 B. hemsleyana were determined against Candida albicans, Escherichia coli, Porphyromonas gingivalis, Staphylococcus aureus and Streptococcus mutans. Scanning electron microscopy (SEM) and bacterial protein leakage assays were used to evaluate its antibacterial activity against P. gingivalis. High-performance liquid chromatography-mass spectrometry (LC-MS) was applied to analyze the ethanol extract of B. hemsleyana 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 P. gingivalis. 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 B. hemsleyana 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 B. hemsleyana has antibacterial and Anti-inflammatory activity. The n-butanol fraction showed a significant antibacterial effect on P. gingivalis.