Abstract
Chemotherapy-induced intestinal mucositis (CIM) is one of the most common side effects of chemotherapy agents. Astragalus licorice prescription (ALP), a traditional Chinese formula, commonly used to treat gastrointestinal disorders, has an unclear mechanism and potential active components in alleviating CIM. This study aims to comprehensively explore the mechanism and bioactive components of ALP in alleviating CIM. ALP's efficacy on CIM was evaluated in Drosophila melanogaster (flies) and C57BL/6 mice using phenotype assays, hematoxylin-eosin (H&E) staining, and immunohistochemistry. ALP's synergistic effect with 5-FU (5-fluorouracil) on tumors was assessed in 615 tumor-bearing mice by measuring tumor volume/weight and performing HE/immunohistochemical staining. Ki67 staining assessed tumor proliferation. Multi-omics integration (transcriptomics, lipidomics, microbiome analysis, network pharmacology) analyzed ALP's mechanism against CIM. Functional pathways were validated via RT-qPCR, biochemical kits, and immunofluorescence, as well as transgenetic flies targeted with GFP. ALP's functional components were characterized by liquid chromatography-mass spectrometry (LC-MS) and validated in CIM flies. ALP significantly mitigated chemotherapy-induced systemic and intestinal damage in flies, evidenced by improved survival rate, elongated intestinal length, reduced acid-base imbalance, and enhanced epithelial and stem cell proliferation. Similarly, ALP alleviated intestinal mucositis symptoms and pathological damage in 5-FU-treated mice, such as reducing diarrhea levels, increasing intestinal length and villus height. Mechanistically, ALP inhibited the expressions of the JAK/STAT pathway related genes (upd3, stat92E, hop, dome, and Dronc) and proteins (UPD3, STAT92E, cleaved caspase-3), and reduced intstinal cells apoptosis. Concurrently, ALP elevated lipid metabolism levels by activating the fatty acid β-oxidation (FAO) pathway related genes expressions (Wdh, Mtp-α, Mtp-β, and Scully) and decreased intestinal free fatty acids. Integrated microbiome, lipidomic, and transcriptomic analyses revealed that ALP corrected multiple gut microbial and lipid metabolic disorders associated with the JAK/STAT apoptotic pathway and FAO lipid metabolism pathway. Furthermore, ALP combined with 5-FU enhanced the anti-tumor effect of 5-FU, as shown by reduced tumor volume and weight, and decreased the proliferation of tumor cells. Finally, four bioactive compounds in ALP, including berberine, dihydrotanshinone I, licochalcone A, and resveratrol, were identified as alleviating CIM. ALP mitigated CIM by inhibiting the JAK/STAT pathway to reduce cellular apoptosis and activating the FAO pathway to improve lipid metabolism, thereby positioning it as a promising novel therapeutic option. Meanwhile, four bioactive compounds of ALP demonstrated protective effects against CIM.
