Integrating Network Pharmacology and Experimental Validation to Elucidate the Anti-neuroinflammatory Mechanism of Hemerocallis citrina Baroni Against Alzheimer’s Disease
Mengjie Xu, Yilin Zhang, Weirong Jie, Jiao Kang, Tingxu YanIntroduction:
Alzheimer's Disease (AD) is a major global health challenge owing to its increasing prevalence, devastating clinical consequences, and the lack of therapeutic interventions. The flower buds of Hemerocallis citrina Baroni (HBF), a traditional herbal medicine and functional food in China, possess diverse neuroprotective activities. This study aimed to investigate the potential pharmacological basis and molecular mechanisms underlying the neuroprotective and anti-AD effects of HBF.
Methods:
The chemical constituents of HBF were characterised using UHPLC-Q-Orbitrap MS/MS. Potential high-affinity ligands, therapeutic targets, and underlying mechanisms were investigated through network pharmacology, molecular docking, and molecular dynamics simulations. Cognitive function was assessed using behavioural tests in AD mice, while histopathological alterations were evaluated by Haematoxylin and Eosin (H&E) and Nissl staining. The effects of HBF on Aβ deposition, tau phosphorylation, NLRP3 inflammasome activation, and inflammatory cytokine expression were further examined using immunohistochemistry, Western blotting, and Enzyme-Linked Immunosorbent Assay (ELISA). In addition, the selective NLRP3 inhibitor MCC950 was employed to evaluate the synergistic effects of HBF.
Results:
Twenty-eight chemical constituents were identified in HBF. Behavioural and histopathological analyses demonstrated that HBF significantly improved cognitive function and alleviated neuronal damage in AD mice. Mechanistically, HBF reduced Aβ deposition and tau hyperphosphorylation, suppressed microglial activation, decreased the expression of proinflammatory cytokines, and inhibited activation of the NF-κB/NLRP3 signalling pathway both in vivo and in vitro. Molecular docking and molecular dynamics simulations further verified strong binding of HBF’s potential high-affinity ligands to NLRP3. Moreover, HBF exhibited synergistic anti-neuroinflammatory effects when combined with the NLRP3 inhibitor MCC950.
Conclusion:
These findings reveal a previously unrecognised neuroprotective mechanism of HBF mediated by the suppression of neuroinflammation through modulation of the NLRP3 inflammasome pathway. This study provides experimental evidence supporting the potential development of HBF as a therapeutic candidate for Alzheimer's disease.