Xin Gao, Shengxia Lv, Yike Wang, Wenyi Zhuang, Mengqi Xiao, Feiye Zhu, Linghui Tao, Yihui Shi, Yongsheng Zhang
Wogonoside and 18β-Glycyrrhetinic acid were identified as candidate bioavailable components, and EPHA7 and LDHB as candidate TCP-responsive targets potentially associated with epithelial-stromal communication, extracellular matrix remodeling, and metabolic regulation. These findings support their functional involvement, although further mechanistic validation is required.
BACKGROUND: Colorectal adenoma (CRA) is a key precancerous lesion, and recurrence or progression remains challenging after endoscopic resection. Shugan Jianpi Tiaochang Pill (TCP), a traditional Chinese medicine formula used for postoperative management, remains insufficiently characterized regarding its bioavailable components and molecular targets.
METHODS: An azoxymethane/dextran sulfate sodium (AOM/DSS)-induced CRA mouse model was used to evaluate TCP. Serum pharmacochemistry identified blood-entering components. Proteomics and network pharmacology were integrated with transcriptomic validation, single-cell and spatial transcriptomics, and virtual knockout analyses to prioritize candidate targets. Molecular docking, molecular dynamics simulations, surface plasmon resonance (SPR), and HCT-116 cell experiments with target-specific knockdown provided validation.
RESULTS: TCP alleviated adenoma-like pathological changes, reduced epithelial proliferation, and showed no obvious hepatic or renal toxicity. Serum pharmacochemistry identified 28 blood-entering components, integrated with proteomics showing partial reversal of disease-associated alterations in mitochondrial energy metabolism. EPHA7 and LDHB were prioritized as candidate TCP-responsive targets. Tissue analyses indicated that TCP restored EPHA7 and LDHB expression toward control levels. Wogonoside and 18β-Glycyrrhetinic acid showed favorable in silico binding and direct SPR binding to EPHA7 and LDHB, with KD values of 4.91-10.3 μM, and inhibited HCT-116 cell viability while promoting apoptosis. EPHA7 or LDHB knockdown partially attenuated the antiproliferative effects of the corresponding compounds.
CONCLUSION: Wogonoside and 18β-Glycyrrhetinic acid were identified as candidate bioavailable components, and EPHA7 and LDHB as candidate TCP-responsive targets potentially associated with epithelial-stromal communication, extracellular matrix remodeling, and metabolic regulation. These findings support their functional involvement, although further mechanistic validation is required.