Molecular Mechanisms of the Anchang Group Prescription in Treating Radiation Enteritis: Network Pharmacology Analysis and Experimental Evidence

Wei Liang ¹ Bo Li ² Yuehong Sun ¹ Dapeng Jia ¹ Tingting Hu ¹ Rujing Huang ¹ Zhilong Liu ¹ Huan Yang ² Baocai Chen ³ Xiaoming Yin ¹ Xinying He ^1* Yun SUN ^1*

• ¹ Cangzhou Hospital of Integrated Traditional Chinese and Western Medicine, Cangzhou, China
• ² School of Pharmacy, North China University of Science and Technology, Tangshan, Hebei Province, China
• ³ Cangzhou Medical College, Cangzhou, Hebei, China

The "Anchang" Group Prescription (ACZF) has demonstrated clinical efficacy in alleviating symptoms of radiation enteritis (RE). This investigation seeks to delineate the active components and explore the molecular mechanisms by which ACZF mitigates RE, utilizing both network pharmacology and experimental approaches .This research constructed a comprehensive "drug-active ingredient-target gene-disease" model leveraging resources such as TCMSP, SwissTargetPrediction, GeneCard, and OMIM. Cytoscape 3.8.2 along with the STRING database were instrumental in developing a protein-protein interaction (PPI) network for the identification of pivotal targets. Functional enrichment analyses were conducted via the DAVID database. Experimentally, a mouse model of RE was induced by X-ray exposure . Parameters measured included body weight, survival rate, incidences of diarrhea, and hematochezia; histological assessments involved hematoxylin and eosin (H&E) and Masson's trichrome staining to examine morphological alterations and collagen deposition in colonic tissues. Levels of cytokines were quantified using enzyme-linked immunosorbent assays (ELISA). Additionally, immunohistochemistry (IHC) and Western blotting (WB) were employed to evaluate the expression of tight junction proteins , as well as proteins linked to the PI3K/AKT pathway. Key bioactive constituents of ACZF in treating RE include quercetin, kaempferol, isorhamnetin, and luteolin, with core target proteins such as SRC, STAT3, AKT. Involved signaling pathways include PI3K/AKT, RAP1, and MAPK. In vivo results revealed that mice treated with ACZF showed enhanced survival, increased body weight, and extended colon lengths compared to controls. H&E staining indicated that radiation-induced mucosal damage, including extensive ulcer formation, inflammatory cell infiltration, crypt structure destruction, and epithelial layer injury, could all be ameliorated by ACZF. Notable reductions were observed in TNF-α, IL-1β, and IL-6 levels, while IL-10 levels saw a significant rise. There was also a marked increase in the expression of ZO-1 and claudin-1. WB analyses demonstrated the activation of the PI3K/AKT pathway in RE, which was significantly curtailed by ACZF, lowering phosphorylation levels within the colonic tissues. Concurrent administration of the PI3K activator YS-49 with ACZF reversed the inhibitory effects on the PI3K/AKT pathway and mitigated impacts on epithelial TJ protein expression and inflammatory cytokine levels, highlighting the critical role of the PI3K/AKT pathway in mediating ACZF’s therapeutic effects in RE.