Astragaloside's Role in Microglial Neuroinflammation: A Multi-Method Study Combining Experimental and Computational Analyses

Abstract

This study investigates the mechanism by which astragaloside (AST) modulates microglial metabolism to alleviate inflammatory injury following oxygen-glucose deprivation/reoxygenation (OGD/R), a model simulating ischemic brain injury. Four groups were studied: Control, Model, AST-treated, and Positive Control (NBP). Cell injury was assessed via CCK-8 assay, and microglial M1 polarization was analyzed by flow cytometry. TNF-α and IL-1β protein levels were measured using ELISA, while IL-1β and IL-10 mRNA expression was quantified by PCR. Core targets were identified through network pharmacology, followed by GO/KEGG enrichment analysis and molecular docking. Protein expression was validated by Western blotting, and metabolic changes were evaluated via metabolomics. AST significantly attenuated OGD/R-induced microglial injury, suppressed TNF-α and IL-1β expression, and inhibited M1 polarization while upregulating IL-10 mRNA. Network pharmacology and molecular docking revealed AST's strong binding affinity to key energy metabolism proteins, including SRC, AKT, and mTOR. Western blotting confirmed activation of SRC and AKT pathways. Metabolomics indicated that AST reversed OGD/R-induced metabolic shifts, downregulating linoleic acid metabolism while modulating glycerophospholipid and pentose phosphate pathways. Astragaloside mitigates OGD/R-induced inflammatory injury by modulating SRC and AKT signaling pathways, leading to metabolic reprogramming in microglia. These findings highlight AST's potential as a therapeutic agent for neuroinflammatory disorders.

Keywords: Astragaloside; Cerebral ischemia–reperfusion injury; Inflammatory response; Metabolism; Microglia polarization.