The Effect and Mechanism of Poria cocos-Atractylodes macrocephala Herb Pair on Chemotherapy-induced Intestinal Mucositis in Mice: A Network Pharmacology and TLR4/MyD88/NF-κB Pathway Study

Abstract: This study aimed to investigate the effect and mechanism of the Poria cocos and Atractylodes macrocephala herb pair on chemotherapy-induced intestinal mucositis (CIM) in mice through network pharmacology prediction and preliminary experimental validation. Network pharmacology was employed to identify potential targets of the herb pair against CIM. Core targets were screened via a protein-protein interaction (PPI) network, and Kyoto Encyclopedia of Genes and Genomes (KEGG) pathway and Gene Ontology (GO) functional analyses were performed on overlapping targets to predict the underlying targets. A CIM mouse model was established by intraperitoneal injection of 5-fluorouracil and intervened with different doses of Poria cocos-Atractylodes macrocephala aqueous decoction or sulfasalazine. Hematoxylin-eosin (HE) staining was used to assess pathological injury in the ileum. Western blot, enzyme-linked immunosorbent assay (ELISA), and immunofluorescence staining were applied to detect relevant target expression and validate network pharmacology predictions. Network pharmacology analysis identified 22 active components of the herb pair and 4993 CIM-related targets, with 111 common targets, including core targets such as Tumor Necrosis Factor (TNF) and SRC. HE staining results demonstrated that the herb pair alleviated ileal pathological damage in CIM mice. ELISA results indicated that the herb pair reduced levels of TNF-α and interleukin-6 (IL-6) in ileal tissue. Western blot and immunofluorescence results showed that the herb pair increased the expression of tight junction proteins (Claudin-1, Occludin, ZO-1) and suppressed activation of the TLR4/MyD88/NF-κB pathway in the ileum. In conclusion, the Poria cocos and Atractylodes macrocephala herb pair ameliorates CIM by inhibiting the TLR4/MyD88/NF-κB pathway, reducing the release of inflammatory cytokines TNF-α and IL-6, and protecting intestinal barrier integrity.