Exploring the mechanism of Cassiae semen in regulating lipid metabolism through network pharmacology and experimental validation

Background: Multiple studies have assessed the role of Cassiae semen (CS) in regulat-ing lipid metabolism. However, the mechanism of action of CS on non-alcoholic fatty liver disease (NAFLD) has seen rare scrutiny. Objective: The objective of this study was to ex-plore the regulatory mechanism of CS on lipid metabolism in NAFLD. Methods: Compo-nents of CS ethanol extract (CSEE) were analyzed and identified using UPLC-Q-Orbirap HRMS. The candidate compounds of CS and its relative targets were extracted from the Traditional Chinese Medicine Systems Pharmacology, Swiss-Target-Prediction, and Target -Net web server. The Therapeutic Target Database, Genecards, Online Mendelian Inheri-tance in Man, and DisGeNET were searched for NAFLD targets. Binding affinity between potential core components and key targets was established employing molecular dock-ing simulations. After that, free fatty acid (FFA)-induced HepG2 cells were used to further validate part of the network pharmacology results. Results: Six genes, including Caspase 3 (CASP3), phosphatidylinositol-4,5-bisphosphate 3-kinase, catalytic subunit alpha (PIK3CA), epidermal growth factor receptor (EGFR), and amyloid beta (A4) precursor protein (APP) were identified as key targets. The mitogen-activated protein kinase (MAPK) signaling pathway was found to associate closely with CS's effect on NAFLD. Per molecular docking findings, toralactone and quinizarin formed the most stable combinations with hub genes. About 0.1 (vs. FFA, P<0.01) and 0.2 (vs. FFA, P<0.05) mg/ml CSEE decreased lipid accumulation in vitro by reversing the up-regulation of CASP3, EGFR, and APP and the down-regulation of PIK3CA. Conclusion: CSEE can significantly reduce intracellular lipid accumulation by modulating the MAPK signaling pathway to decrease CASP3 and EGFR expression.

Automated summary

This study explored the regulatory mechanism of Cassiae semen (CS) on lipid metabolism in non-alcoholic fatty liver disease (NAFLD) using network pharmacology and in vitro experiments.