Network pharmacology identifies the mechanisms of action of Jianpi Yanggan Pill in treating diarrhea-predominant irritable bowel syndrome.

OBJECTIVE

To investigate the molecular mechanisms of Jianpi Yanggan Pill (JPYGP) in treating Diarrhea-predominant irritable bowel syndrome (IBS-D) using an integrated network pharmacology approach combined with experimental validation.

METHODS

The chemical constituents of JPYGP were identified by ultra-high performance liquid chromatography-high-resolution mass spectrometry (UHPLC-MS/MS), and bioactive compounds were screened using various databases. Candidate targets was predicted, followed by Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway enrichment studies. An IBS-D rat model was established using acetic acid enema combined with restraint stress. The therapeutic potential of JPYGP was further validated through biochemical assays, including Western blotting and enzyme-linked immunosorbent assay (ELISA), to assess the involvement of the C-X-C motif chemokine ligand 8 and extracellular signal-regulated kinase (CXCL8-ERK) signaling pathway.

RESULTS

UHPLC-MS/MS analysis identified 2,309 candidate compounds in JPYGP, among which 20 were bioactive. A total of 660 targets were predicted, of which 414 overlapped with IBS-D-related targets. Enrichment analysis highlighted that these targets were mainly involved in inflammatory and MAPK pathways. In a living organism, JPYGP noticeably alleviated diarrhea symptoms and visceral pain, reduced colonic tissue damage, inhibited mast cell activation, and downregulated the expression of CXCL8 and phosphorylated ERK1/2.

CONCLUSION

JPYGP exerts therapeutic effects on IBS-D via a multi-component, multi-target mechanism, primarily by suppressing the CXCL8-ERK pathway, thereby attenuating colonic inflammation and visceral hypersensitivity.