Huayu Xiaopi Decoction regulates the JNK/c-Jun/Slug pathway to inhibit epithelial-mesenchymal transformation and improve precancerous lesions of gastric cancer

Abstract

Ethnopharmacological significance: Huayu Xiaopi Decoction (HYXPT), a traditional Chinese medicine formula, exhibits favorable therapeutic effects on precancerous lesions of gastric cancer (PLGC). However, a comprehensive characterization of its chemical constituents and absorbed components remains elusive, and the mechanisms underlying its efficacy in treating PLGC warrant further investigation.

Aim of the study: To explore the therapeutic effects and underlying mechanisms of HYXPT in the treatment of PLGC through an integrated approach combining bioinformatics analyses with in vivo and in vitro experiments.

Materials and methods: The UHPLC-Q-Exactive Orbitrap MS/MS method was used to analyze the active components in HYXPT water extract. Subsequently, network pharmacology and molecular docking analyses were used to identify the relevant pathways. In vivo, a PLGC rat model was established through five-factor composite modeling with MNNG. In vitro, a malignant transformed cell model was induced in GES-1 cells using MNNG to observe and validate the mechanism of HYXPT intervention in PLGC progression.

Results: A total of 574 active components and 44 active components were successfully identified in the HYXPT water extract and rat serum, respectively. Network pharmacology identified 182 potential targets. Protein-protein interaction (PPI) network analysis showed that the core targets of HYXPT treatment for PLGC were TP53, AKT1, and Jun. GO and KEGG enrichment analysis showed that the therapeutic effects of HYXPT on PLGC were related to the MAPK signaling pathway. Molecular docking results revealed that the active components of HYXPT demonstrated good binding affinity with Jun. In vivo experiments demonstrated that HYXPT alleviated gastric histological injury in PLGC rats in a dose-dependent manner. The protein expression levels of p-JNK, p-c-Jun, Slug, N-cadherin, and Vimentin decreased significantly in gastric tissue, whereas the protein content of E-cadherin increased significantly. In vitro experiments showed that HYXPT, similar to the JNK inhibitor SP600125, could reduce the proliferation and migratory activity of MC cells. Consistent with in vivo findings, the protein expressions of p-JNK, p-c-Jun, Slug, N-cadherin and Vimentin in MC cells decreased significantly, while E-cadherin protein expression increased significantly.

Conclusion: HYXPT exerts its therapeutic effects on PLGC by regulating the JNK/c-Jun/Slug signaling pathway, thereby inhibiting EMT during PLGC progression.

Keywords: Bioinformatics; Epithelial mesenchymal transformation; Huayu Xiaopi Decoction; JNK/c-Jun/Slug; Precancerous lesions of gastric cancer.