Exploring the potential mechanisms of piperine against Alzheimer’s disease through network pharmacology, molecular docking, and in vitro experiments

Abstract

Introduction: Alzheimer’s disease (AD) is a complex neurodegenerative disorder with limited treatment options, prompting interest in natural compounds with multi-target activity. Piperine has shown neuroprotective and anti-inflammatory potential, but its mechanisms against AD have not been fully elucidated.

Objective: This study employed a combined strategy of network pharmacology, molecular docking, and in vitroassays to explore the potential mechanisms by which piperine may modulate AD-related neuroinflammation.

Methods: Potential targets of piperine and AD-related targets were retrieved from multiple public databases. Common targets were analyzed via protein-protein interaction (PPI) network, Gene Ontology, and Kyoto Encyclopedia of Genes and Genomes (KEGG) enrichment analyses, and binding affinities were evaluated using molecular docking. For in vitrovalidation, BV2 microglial cells were stimulated with lipopolysaccharide (LPS) and treated with piperine. Cytokine (tumor necrosis factor alpha [TNF-α], IL [interleukin]-18, IL-1β) levels were measured by ELISA, and mRNA expression of key targets was assessed by qRT-PCR.

Results: A total of 134 overlapping targets were identified between piperine and AD. Core targets, including AKT1, HSP90AA1, SRC, RELA, and MAPK1, were discerned from the PPI network. KEGG enrichment analysis implicated several key pathways, such as PI3K-Akt, apoptosis, and cAMP signaling. Molecular docking simulations predicted stable binding interactions between piperine and these core targets, with binding energies all below −5.0 kcal/mol. In cellular experiments, piperine significantly inhibited the LPS-induced release of TNF-α, IL-18, and IL-1β. At the transcriptional level, piperine downregulated the mRNA expression of Src, Rela, and Mapk1while upregulating the expression of Akt1and Hsp90aa1(p< 0.05).

Conclusion: These findings suggest that piperine alleviates neuroinflammation in a microglial model, potentially by modulating a network of targets (AKT1, HSP90AA1, SRC, RELA, and MAPK1)and pivotal pathways, supporting the continued assessment of piperine as a potential anti-neuroinflammatory agent relevant to AD.

Keywords

Piperine / Alzheimer’s disease / Network pharmacology approach / Molecular docking / Neuroinflammation / BV2 microglial cells

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