Abstract
To explore the mechanism of sophora flavescens, cortex fraxini, and pomegranate peel complex powder (Compound Kushen Powder) in the treatment of animal diarrhea, a network pharmacology approach leveraging databases like TCMSP and SwissTarget was applied in this study. Molecular docking was executed between the primary constituents and pivotal targets, enabling an additional refinement of main targets and key medications. Subsequently, a rat diarrhea model induced by folium sennae leaves was established for in vivo validation. The rats were divided into four groups: negative control group, positive control group, positive drug treatment group, and Compound Kushen Powder treatment group. Key protein targets, such as Caspase-3, IL-1β, IL-10, MMP9, STAT3, TNF, TP53, and VEGFA, essential for mitigating diarrhea in response to the composite medication were found through network pharmacology. Additionally, the results of molecular docking analysis unveiled fundamental constituents of Compound Kushen Powder, namely beta-sitosterol, ursolic acid, formononetin, and matrine, which demonstrated significant binding affinities with those identified key protein targets. The results of mRNA and protein expression analyses of rat colonic tissue validated the in vivo alterations of core genes identified through network screening. Except for IL-10 and STAT3, the expression of all targets exhibited noteworthy reductions when compared to the positive control group (P < 0.05). These results demonstrated that Compound Kushen Powder can inhibit inflammation and regulate cell apoptosis by modulating signaling pathways such as IL-17, TNF-α, MAPK, and NF-κB. Collectively, this study sheds light on the traditional application of complex powder for the prevention and treatment of diarrhea.