关键词:

"> 化橘红, 高脂血症, 网络药理学, 分子对接, HepG2 细胞

Abstract:

This study aims to explore the mechanism of Citri Grandis Exocarpium (CGE) in the treatment of hyperlipidemia based on network pharmacology and experimental verification. The active components and potential targets of CGE in the treatment of hyperlipidemia were screened through databases such as TCMSP and STRING. The core targets were analyzed through GO functional annotation and KEGG pathway enrichment. The protein-protein interaction network was constructed using the STRING database. Molecular docking technology was used to simulate the binding ability between the active compounds of CGE and the key target proteins. Furthermore,an oleic acid-induced HepG2 cell model was established,and Oil Red O staining,triglyceride (TG) and total cholesterol (TC) assays,as well as Western blot analysis were used to verify the lipid-lowering effect of CGE. After screening and analysis,20 potential targets of CGE were obtained. The core active compounds included naringenin,isosinensetin,and apigenin. The key targets related to the treatment of hyperlipidemia mainly included cysteine-aspartic acid protease 3 ( CASP3),serine/ threonine kinase 1 ( AKT1),and prostaglandin-endoperoxide synthase 2 (PTGS2). GO enrichment analysis indicated that the potential targets of CGE were mainly involved in biological processes such as apoptosis,response to lipopolysaccharide,and protein phosphorylation. KEGG pathway enrichment suggested that CGE was closely related to metabolic pathways and the phosphoinositide 3-kinase/ protein kinase B (PI3K/ AKT) signaling pathway. Molecular docking results showed that the core components exhibited favorable binding activities with the key targets,among which naringenin with CASP3 (-6. 5 kcal/ mol) and apigenin with AKT1 (-6. 15 kcal/ mol) showed the best binding energies. In vitro experiments showed that 0. 2 mmol/ L oleic acid successfully induced lipid accumulation in HepG2 cells, while CGE treatment reduced intracellular lipid droplets in a dose-dependent manner,significantly decreasing the Oil Red O positive area from 39. 89% to 19. 4% -23. 92% ,and markedly lowering TG and TC levels. In addition,CGE inhibited lipid synthesis by suppressing the PI3K/ AKT signaling pathway. This study reveals that CGE alleviates lipid accumulation in HepG2 cells by regulating lipid metabolism-related signaling pathways,which provides a theoretical basis for further investigation of its lipid-lowering mechanism.

Key words:

"> Citri Grandis Exocarpium, hyperlipidemia, network pharmacology, molecular docking, HepG2 cells