Inosine, AMP, and Vidarabine: Network Pharmacology and LC-MS Reveal Key Bioactive Compounds in Periplaneta americana for Ulcerative Colitis Management

Abstract

Ulcerative colitis (UC) is a chronic inflammatory bowel disease with unmet therapeutic needs. This study investigates the therapeutic potential of Periplaneta americana L. extract (PAE) and its molecular mechanisms, integrating network pharmacology and experimental validation. Liquid chromatography-mass spectrometry identified 1355 compounds in PAE. Network pharmacology analysis revealed that inosine, vidarabine, and adenosine 5'-monophosphate (AMP) were core components and the core components synergistically regulated key targets and acted on inflammation-related pathways, thereby establishing a multi-target anti-inflammatory regulatory network. In vivo experiments demonstrated that these compounds significantly alleviated colitis symptoms in dextran sulfate sodium-induced mice, as evidenced by reduced disease activity index scores, preserved colonic mucosal architecture, and decreased inflammatory infiltration. Mechanistically, core compounds down-regulated granulocyte-macrophage colony-stimulating factor (GM-CSF), inducible nitric oxide synthase (iNOS)/NOS2, monocyte chemoattractant protein 1 (MCP-1), and transforming growth factor beta 1 (TGF-β1), while they up-regulated interleukin-10 (IL-10) and epidermal growth factor (EGF). Additionally, they activated epidermal growth factor receptor (EGFR)-mediated pathways. Molecular docking analysis revealed that adenosine analogs preferentially bound to A1/A2a receptors, triggering signaling cascades essential for epithelial repair and inflammation resolution. This study established the multi-component, multi-pathway mechanism of PAE in UC, highlighting its dual role in suppressing inflammation and promoting mucosal healing. By bridging traditional herbal use with modern molecular insights, these findings provided a translational foundation for developing PAE-based therapies for UC.

Keywords: Periplaneta americana; adenosine analogs; inflammation; mucosal repair; network pharmacology; ulcerative colitis.

Figure 1

LC-MS/MS chromatogram of PAE. (A) Total ion chromatogram (TIC) of PAE. (B) The enlarged view of the range from 0.5 to 6 min in the TIC.

Figure 2

Potential targets of PAE in the treatment of UC. (A) Overlap between targets of G1 compounds and disease/symptom targets; (B) Overlap between targets of G2 compounds and disease/symptom targets; (C) KEGG enrichment analysis of overlapping genes between drug targets and disease targets (left: G1, right: G2); (D) Network of active components, targets, and pathways.

Figure 3

Venn diagram of intersecting signaling pathways of G1 (A), G2 (B) compounds for the treatment of diseases/symptoms.

Figure 4

Molecular docking results of key targets with main components in PAE. (A) Heatmap of binding energies from molecular docking between core compounds and potential targets. (B) Molecular docking of adenosine 2′,3′-cyclic phosphate with ADORA2a. (C) Molecular docking of adenosine 5′-monophosphate with ADORA1. (D) Molecular docking of adenosine 5′-monophosphate with ADORA2a.

Figure 5

Effects of PAE on colitis symptoms. (A) Body weight changes in UC mice during administration. (B) Colonic morphology of mice. (C) Disease activity index (DAI) scores during administration. (D) Microscopic observation of H&E-stained section, red arrow: Inflammation, green arrow: Crypt destruction. (E) Levels of pro-inflammatory cytokines (GM-CSF, EGF, IL-10, IL-17A, MCP-1, TGF-β1, and iNOS/NOS2) assessed using ELISA kits. All data are presented as mean ± SD (n = 6 per group); * p < 0.05, *** p < 0.001 vs. NC group; ** p < 0.01, *** p < 0.001 vs. MC group.

Figure 6

PAE restores the intestinal barrier in DSS-induced colitis. (A) Expression levels of EGFR, ERK, PI3K, AKT, and Src in colonic tissues assessed by immunohistochemical (IHC) analysis (positive staining is indicated by pale yellow, brown, or tan granules). (B) Quantification of IHC results (20× magnification; scale bar: 50 µm). Data are expressed as mean ± SD; ** p < 0.01, *** p < 0.001 vs. NC group; ** p < 0.01, *** p < 0.001 vs. MC group.