Potential improvement of Isaria cicadae Miquel spores oil(ICSO)alleviate TNBS-induced fibrosis of Caco-2 cells by down-regulating the activation of EGFR and SRC in the MEK/ERK and PI3K/Akt pathways

The natural oil extracted from Isaria cicadae Miquel (ICSO), a rare mushroom species, has demonstrated efficacy in alleviating intestinal fibrosis associated with Crohn's disease (CD), although its precise mechanisms remain unclear. This study employed an integrated approach combining network pharmacology, molecular docking, and experimental validation to investigate ICSO's therapeutic potential. ICSO obtained through Soxhlet extraction was characterized by GC–MS, revealing 21 components including 16 fatty acids (84.82 % of total composition). In a TNBS-induced Caco-2 cell fibrosis model, ICSO (20-200 μg/mL) dose-dependently attenuated intestinal fibrosis, significantly reducing hydroxyproline accumulation ( P < 0.05) and suppressing type I and II collagen deposition ( P < 0.01). Furthermore, Consistent with network pharmacology predictions, ICSO (50, 100, and 200 μg/mL) markedly inhibited EGFR overexpression, suppressed SRC activation, while downregulating AKT ( P < 0.001) and MAPK1 phosphorylation ( P < 0.05). These coordinated mechanisms effectively ameliorated epithelial-mesenchymal transition (EMT) and prevented intestinal fibrosis progression in CD. • Network pharmacology and molecular docking predict key targets of ICSO in Crohn's disease fibrosis. • ICSO improves the excessive deposition of collagen types I and III and reduces the accumulation of extracellular matrix. • ICSO can effectively reduce the mesenchymalization of epithelial cells, thus preventing the occurrence of fibrosis. • ICSO slows down fibrosis development by down-regulating EGFR and inhibiting P13K/AKT and MEK/ERK signaling.