Integrated analysis strategies reveal the pharmacodynamic substances and action mechanisms of Huangqi San in the treatment of Alzheimer's disease.

Alzheimer's disease (AD), as a neurodegenerative disorder characterized primarily by memory impairment in its early stages, imposes a heavy burden on society and medical resources. This study aims to investigate the therapeutic effect of Huangqi San (HQS) on AD and elucidate its potential mechanism of action. This study analyzed the rat serum after treated by HQS using UHPLC-Q-Exactive Orbitrap MS, and identified the potential active components of HQS. Subsequently, key targets and pathways were identified through network pharmacology and molecular docking techniques, and were verified in PC12 cells damaged by Aβ25-35. The verification methods included cell viability assays, analysis of inflammatory/oxidative stress markers, and regulation of the PI3K/AKT/MAPK pathway. The results showed that a total of 40 serum adsorption components and 70 common targets were identified. Network analysis revealed that the PI3K-AKT, MAPK and Rap1 signaling pathways were at the core of the network. Molecular docking demonstrated strong binding affinity between 10 core components (e.g., isoliquiritigenin, moracin M) and pivotal targets (BCL2, MAPK3, PTGS2). In vitro validation showed HQS significantly attenuated Aβ25-35-induced neurotoxicity by enhancing cell viability, suppressing apoptosis, reducing pro-inflammatory cytokines (TNF-α, IL-1β, IL-6) and oxidative markers (MDA, ROS, CYP2E1) while elevating GSH-Px activity. Moreover, HQS can bidirectionally regulate PI3K/AKT/MAPK pathways via upregulating PI3K/AKT/ERK1/2 and inhibiting JNK/p38 phosphorylation. In conclusion, this study innovatively established HQS as a multi-component/multi-target candidate drug for the treatment of AD. Its mechanism of action is related to its synergistic regulation of PI3K/AKT/MAPK-mediated neuroprotection. This comprehensive approach provides an example for interpreting complex traditional Chinese herbal preparations.