Exploration of Berberine Against Ulcerative Colitis via TLR4/NF-κB/HIF-1α Pathway by Bioinformatics and Experimental Validation

Drug Des Devel Ther. 2024 Jul 9:18:2847-2868. doi: 10.2147/DDDT.S436359. eCollection 2024.

Abstract

Purpose: This study aimed to delineate the molecular processes underlying the therapeutic effects of berberine on UC by employing network pharmacology tactics, molecular docking, and dynamic simulations supported by empirical validations both in vivo and in vitro.

Patients and methods: We systematically screened potential targets and relevant pathways affected by berberine for UC treatment from comprehensive databases, including GeneCards, DisGeNET, and GEO. Molecular docking and simulation protocols were used to assess the interaction stability between berberine and its principal targets. The predictions were validated using both a DSS-induced UC mouse model and a lipopolysaccharide (LPS)-stimulated NCM460 cellular inflammation model.

Results: Network pharmacology analysis revealed the regulatory effect of the TLR4/NF-κB/HIF-1α pathway in the ameliorative action of berberine in UC. Docking and simulation studies predicted the high-affinity interactions of berberine with pivotal targets: TLR4, NF-κB, HIF-1α, and the HIF inhibitor KC7F2. Moreover, in vivo analyses demonstrated that berberine attenuates clinical severity, as reflected by decreased disease activity index (DAI) scores, reduced weight loss, and mitigated intestinal inflammation in DSS-challenged mice. These outcomes include suppression of the proinflammatory cytokines IL-6 and TNF-α and downregulation of TLR4/NF-κB/HIF-1α mRNA and protein levels. Correspondingly, in vitro findings indicate that berberine decreases cellular inflammatory injury and suppresses TLR4/NF-κB/HIF-1α signaling, with notable effectiveness similar to that of the HIF-1α inhibitor KC7F2.

Conclusion: Through network pharmacology analysis and experimental substantiation, this study confirmed that berberine enhances UC treatment outcomes by inhibiting the TLR4/NF-κB/HIF-1α axis, thereby mitigating inflammatory reactions and improving colonic pathology.

Keywords: TLR4/NF-κB/HIF-1α; berberine; experimental validation; inhibitor; molecular docking; network pharmacology; ulcerative colitis.

MeSH terms

  • Animals
  • Berberine* / chemistry
  • Berberine* / pharmacology
  • Colitis, Ulcerative* / chemically induced
  • Colitis, Ulcerative* / drug therapy
  • Colitis, Ulcerative* / metabolism
  • Colitis, Ulcerative* / pathology
  • Computational Biology*
  • Dextran Sulfate
  • Disease Models, Animal
  • Humans
  • Hypoxia-Inducible Factor 1, alpha Subunit* / antagonists & inhibitors
  • Hypoxia-Inducible Factor 1, alpha Subunit* / metabolism
  • Lipopolysaccharides / antagonists & inhibitors
  • Lipopolysaccharides / pharmacology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Molecular Docking Simulation
  • NF-kappa B* / antagonists & inhibitors
  • NF-kappa B* / metabolism
  • Network Pharmacology
  • Toll-Like Receptor 4* / antagonists & inhibitors
  • Toll-Like Receptor 4* / metabolism

Substances

  • Berberine
  • Toll-Like Receptor 4
  • Hypoxia-Inducible Factor 1, alpha Subunit
  • NF-kappa B
  • TLR4 protein, human
  • Lipopolysaccharides
  • HIF1A protein, human
  • Dextran Sulfate
  • Tlr4 protein, mouse
  • Hif1a protein, mouse

Grants and funding

This study was supported by National Natural Science Foundation of China (82174304).