Effect of Gynostemma pentaphyllum on Ulcerative Colitis by Regulating the PI3K‐AKT Signaling Pathway

This study employed a methodology that integrates network pharmacology predictions with in vitro cellular experiments to preliminarily investigate the pharmacological mechanisms of Gynostemma pentaphyllum (GP) in the treatment of ulcerative colitis (UC). Through comprehensive database searches, 15 active compounds of GP were identified, encompassing 173 potential targets, while 4218 UC‐related targets were recognized, resulting in the identification of 136 common targets. A protein interaction network was constructed utilizing the STRING database, and Gene Ontology (GO) and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses were conducted using the Metascape database. Molecular docking validation was performed using AutoDock software to assess the interactions between the primary active components and core targets. In vitro experiments were conducted using a Caco‐2 cell model induced by lipopolysaccharide (LPS), where cell viability was assessed via the cell counting kit‐8 (CCK‐8) assay, and levels of IL‐6 and TNF‐α were measured using ELISA. Additionally, the expression of total PI3K and Akt proteins was analyzed through western blotting. The findings of this study indicate that network pharmacology analysis identified 10 core targets, including AKT1, TNF, IL6, TP53, and IL1B. KEGG enrichment analysis suggested the involvement of signaling pathways such as PI3K‐Akt, MAPK, TNF, and IL‐17 in the intervention process of GP in UC. Molecular docking demonstrated that the primary active components (rusine, quercetin, and isorhizanol) exhibited strong binding affinities with target proteins, including AKT1, TNF, IL6, and TP53. Cellular experiments revealed that single‐dose GP treatment resulted in decreased levels of IL‐6 and TNF‐α in LPS‐induced Caco‐2 cells, alongside alterations in the total protein expression of PI3K and Akt. It is important to note that this study did not assess phosphorylated proteins, rendering it unable to ascertain the activity state of the PI3K‐Akt pathway. Furthermore, the use of a single cell line and a single dose limits the extrapolation of these findings to in vivo contexts. In conclusion, the potential therapeutic effect of GP on UC may involve a complex mechanism encompassing multiple targets and pathways. This study provides preliminary in vitro experimental evidence and formulates testable scientific hypotheses for further in‐depth investigation.