基于血清代谢组学与网络药理学探讨橙黄决明素抗动脉粥样硬化的作用机制OA

Objective To investigate the mechanism of aurantio-obtusin against atherosclerosis(AS)based on serum metabolomics and network pharmacology.Methods(1)An AS mouse model was established in ApoE-/-mice(C57BL/6 background)fed a high-fat diet for 24 weeks.Wild-type C57BL/6J mice were used as the control group,ApoE-/-mice were randomly divided into model group,low-dose aurantio-obtusin group(10 mg·kg-1·d-1),and high-dose aurantio-obtusin group(20 mg·kg-1·d-1)(n=6 per group).After 8 weeks of high-fat diet intervention,the low-dose and high-dose aurantio-obtusin groups received the drug by intragastric gavage once daily for 16 consecutive weeks.Histopathological changes in the aortic sinus were observed by HE staining,Masson staining,and Oil Red O staining.(2)Serum non-targeted metabolomics analysis was performed using LC-MS/MS.Orthogonal partial least squares-discriminant analysis(OPLS-DA)was employed to validate metabolic profile differences among groups and screen differential metabolites.Pathway enrichment analysis of differential metabolites was conducted.(3)Potential targets of aurantio-obtusin against AS were screened by network pharmacology.A protein-protein interaction(PPI)network was constructed,and GO function and KEGG pathway enrichment analyses were performed.Correlation analysis between network pharmacology and metabolomics results was conducted to construct a"drug-target-pathway-metabolite"network.Molecular docking was performed to validate the binding affinity between aurantio-obtusin and key targets.Results(1)Compared with the control group,the model group showed significantly increased plaque area in the aortic sinus(P＜0.01),massive lipid deposition(P＜0.01),and increased collagen fiber content in the aortic plaque area(P＞0.05).Compared with the model group,both low-dose and high-dose aurantio-obtusin groups exhibited significantly reduced plaque area in the aortic sinus(P＜0.01),significantly decreased lipid accumulation(P＜0.01),and significantly increased collagen deposition within plaques(P＜0.01).(2)Aurantio-obtusin significantly modulated the serum metabolic profile of AS mice.A total of 32 significantly differential metabolites were identified in the high-dose aurantio-obtusin group vs.the model group,among which 24 were upregulated and 8 were downregulated.These differential metabolites were mainly enriched in pathways including steroid hormone biosynthesis,mTOR signaling pathway,and PPAR signaling pathway.(3)Network pharmacology analysis identified 131 potential targets of aurantio-obtusin in AS,mainly involved in biological processes such as collagen catabolic process and enriched in key signaling pathways including lipid and atherosclerosis pathway,fluid shear stress and atherosclerosis pathway,and PI3K-Akt signaling pathway.Correlation analysis further identified 5 key targets directly associated with serum differential metabolites:NOS3,ARG1,ALDH2,CYP2C9,and CYP19A1,and 4 key metabolites:L-arginine,indole-3-acetic acid,3-methoxy-4-hydroxymandelic acid,and 8-hydroxy-eicosatetraenoic acid.Aurantio-obtusin exhibited favorable binding affinity with all 5 key target proteins.Conclusion Aurantio-obtusin can significantly ameliorate pathological changes in the aortic sinus of AS mice,potentially by targeting key proteins such as NOS3,ARG1,ALDH2,CYP2C9,and CYP19A1,and modulating the L-arginine/NO metabolic axis,tryptophan metabolism pathway,tyrosine metabolism pathway,and arachidonic acid metabolic network,thereby achieving multi-coordinated regulation of vascular endothelial function,inflammatory response,and lipid metabolism to exert its anti-AS effects.