Exploring the Mahuang Fuzi Xixin Decoction's Mechanism for Treating Alzheimer's Disease (AD) Using Molecular Docking and Network Pharmacology

Che Chen^1*Qianfeng Shao²Sheng Zhou³

• ¹Ningxia Medical University, Yinchuan, China
• ²Shenzhen Bao'an Hospital of Traditional Chinese Medicine, Shenzhen, China
• ³Qingdao Ruyi Software Co., Ltd., Medical Data Analysis Center, Qingdao, China, Qingdao, China

Objective: To explore the potential mechanism of Mahuang Fuzi Xixin Decoction (MFXD) in treating Alzheimer's disease (AD) via network pharmacology, molecular docking, and test its efficacy by in vitro experiments. Methods: Active components of MFXD were screened from TCMSP, BATMAN-TCM, TCMID, with targets obtained from SwissTargetPrediction and TCMSP. AD-related differential genes were retrieved from GEO. Intersection targets were identified via Venn diagrams, followed by GO/KEGG enrichment, PPI network construction, and molecular docking. In vitro experiments used Aβ₂₅₋₃₅-induced PC12 cells to simulate AD pathology. CCK-8 assay evaluated cell viability; Western blot detected key proteins (p-NF-κB p65, GSK-3β, MMP-9, Tau, etc.); ELISA measured TNF-α secretion to assess anti-inflammatory effects. Results: Thirty-seven active components and 230 targets of MFXD were identified, along with 4913 AD differential genes from GSE122063, yielding 47 intersection targets. GO/KEGG enrichment involved reactive oxygen species metabolism, TNF and NF-κB pathways; core targets included MMP9, EGFR, FOS, with quercetin and luteolin showing good binding. In vitro, quercetin/luteolin increased cell viability concentration-dependently (all P < 0.001); 15% MFXD-containing serum restored viability to ≥95% (P < 0.001 vs. AD model, comparable to DHCL). Western blot showed AD model had abnormal protein levels (all P < 0.05); MFXD reversed these, while DHCL only inhibited p-NF-κB p65/NF-κB p65. ELISA showed both reduced TNF-α (all P < 0.001). Conclusion: MFXD potentially exerts anti-AD effects through a multi-component, multi-target, multi-pathway approach. Key components (quercetin, luteolin) may modulate core target MMP9. It also regulates TNF, Calcium, NF-κB pathways, targets Tau pathology by restoring GSK-3β phosphorylation to suppress abnormal Tau hyperphosphorylation, alleviating AD pathological damage.