AccScience Publishing / EJMO / Volume 10 / Issue 3 / DOI: 10.36922/EJMO026030033
Submit to EJMO
Cite this article
3
Download
27
Views
Related Info Links
Google Scholar
More by Authors Links
Kok Hong Tang
Journal Browser
Volume | Year
Issue
Search
Forthcoming Issue
Current Issue
View All
News and Announcements
View All
ORIGINAL RESEARCH ARTICLE

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

Shangzhi Wu1 ,  Kok Hong Tang1*
EJMO 2026, 10(3), 026030033 https://doi.org/10.36922/EJMO026030033
Received: 16 January 2026 | Revised: 8 April 2026 | Accepted: 9 April 2026 | Published online: 29 June 2026
© 2026 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution -Noncommercial 4.0 International License (CC-by the license) ( https://creativecommons.org/licenses/by-nc/4.0/ )
Download PDF
XML
Cite
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 vitro assays 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 vitro validation, 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 Mapk1 while upregulating the expression of Akt1 and 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
Funding
The funding is provided by Guangxi University of Chinese Medicine (grant no.: 2020GXNSFAA297179).
Conflict of interest
The authors declare no conflict of interest.
References
  1. Agrawal M, Saraf S, Saraf S, et al. Nose-to-brain drug delivery: an update on clinical challenges and progress towards approval of anti-Alzheimer drugs. J Control Release. 2018;281:139-177. doi: 10.1016/j.jconrel.2018.05.011

 

  1. Prince M, Bryce R, Albanese E, et al. The global prevalence of dementia: a systematic review and metaanalysis. Alzheimers Dement. 2013;9(1):63-75.e2. doi: 10.1016/j.jalz.2012.11.007

 

  1. Singh G, Chauhan P, Wadhwa K. Unravelling the Alzheimer’s pathogenesis: molecular and cellular pathways to neurodegeneration and therapeutic targets. Eur J Pharmacol. 2025;983:177123. doi: 10.1016/j.prp.2025.156109

 

  1. Zhang T, Pan L, Cao Y, et al. Identifying the mechanisms and molecular targets of Yizhiqingxin formula on Alzheimer’s disease: coupling network pharmacology with GEO database. Pharmgenomics Pers Med. 2020;13:487-502. doi: 10.2147/PGPM.S269726

 

  1. Gorgani L, Mohammadi M, Najafpour GD, et al. Piperine— the bioactive compound of black pepper: from isolation to medicinal formulations. Compr Rev Food Sci Food Saf. 2017;16(1):124-140. doi: 10.1111/1541-4337.12246

 

  1. Tripathi AK, Ray AK, Mishra SK. Molecular and pharmacological aspects of piperine as a potential molecule for disease prevention and management: evidence from clinical trials. Beni Suef Univ J Basic Appl Sci. 2022;11(1):16. doi: 10.1186/s43088-022-00196-1

 

  1. Wang C, Cai Z, Wang W, et al. Piperine attenuates cognitive impairment in an experimental mouse model of sporadic Alzheimer’s disease. J Nutr Biochem. 2019;70:147-155. doi: 10.1016/j.jnutbio.2019.04.009

 

  1. Azramezani Kopi T, Jafari A, Mirzaei M, et al. Piperine prevents scopolamine-induced cognitive impairment via its antioxidant and anti-inflammatory roles; suggesting potential modulation of necroptosis-related genes including MLKL and TNF-α. J Mol Neurosci. 2025;75(4):142. doi: 10.1007/s12031-025-02437-1

 

  1. Head E, Murphey HL, Dowling AL, et al. A combination cocktail improves spatial attention in a canine model of human aging and Alzheimer’s disease. J Alzheimers Dis. 2012;32(4):1029-1042. doi: 10.3233/JAD-2012-120937

 

  1. Nazifi M, Oryan S, Esfahani DE, et al. The functional effects of piperine and piperine plus donepezil on hippocampal synaptic plasticity impairment in rat model of Alzheimer’s disease. Life Sci. 2021;265:118802. doi: 10.1016/j.lfs.2020.118802

 

  1. Bhanukiran K, Singh SK, Singh R, et al. Discovery of multitarget-directed ligands from piperidine alkaloid piperine as a cap group for the management of Alzheimer’s disease. ACS Chem Neurosci. 2023;14(15):2743-2760. doi: 10.1021/acschemneuro.3c00257

 

  1. Aktary N, Jeong Y, Oh S, et al. Unveiling the therapeutic potential of natural products in Alzheimer‘s disease: insights from in vitro, in vivo, and clinical studies. Front Pharmacol. 2025;16:1601712. doi: 10.3389/fphar.2025.1601712

 

  1. Müller A, Schmidt S, Wagner H. Network pharmacology to elucidate the role of phytotherapy in neurocognitive disorders. Phytomedicine. 2025;148:157031. doi: 10.1016/j.phymed.2025.157031

 

  1. Ngo T, Kim B, Lee H, Park M. Leveraging network pharmacology for drug discovery: integrative approaches and emerging insights. Med Drug Discov. 2025;27:100220. doi: 10.1016/j.medidd.2025.100220

 

  1. Yu Z, Wu Z, Wang Z, et al. Network-based methods and their applications in drug discovery. J Chem Inf Model. 2024;64(1):57-75. doi: 10.1021/acs.jcim.3c01613

 

  1. Gfeller D, Grosdidier A, Wirth M, et al. SwissTargetPrediction: a web server for target prediction of bioactive small molecules. Nucleic Acids Res. 2014;42(W1):W32-W38. doi: 10.1093/nar/gku293

 

  1. Li Y, Jiang Y, Zhu B, et al. Unveiling the potential targets and mechanisms of DEHP/BPA exposure on PCOS: insights from network toxicology and molecular docking. J Ovarian Res. 2025;18(1):268. doi: 10.1186/s13048-025-01866-1

 

  1. Szklarczyk D, Gable AL, Lyon D, et al. STRING v11: protein-protein association networks with increased coverage, supporting functional discovery in genome-wide experimental datasets. Nucleic Acids Res. 2019;47(D1):D607-D613. doi: 10.1093/nar/gky1131

 

  1. Shannon P, Markiel A, Ozier O, et al. Cytoscape: a software environment for integrated models of biomolecular interaction networks. Genome Res. 2003;13(11):2498-2504. doi: 10.1101/gr.1239303

 

  1. Ogata H, Goto S, Sato K, et al. KEGG: Kyoto Encyclopedia of Genes and Genomes. Nucleic Acids Res. 1999;27(1):29-34. doi: 10.1093/nar/27.1.29

 

  1. Cheng M, Li M, Zhang Y, et al. Exploring the mechanism of PPCPs on human metabolic diseases based on network toxicology and molecular docking. Environ Int. 2025;196:109324. doi: 10.1016/j.envint.2025.109324

 

  1. Trott O, Olson AJ. AutoDock Vina: improving the speed and accuracy of docking with a new scoring function, efficient optimization, and multithreading. J Comput Chem. 2010;31(2):455-461. doi: 10.1002/jcc.21334

 

  1. Shah-Abadi ME, Ariaei A, Moradi F, et al. In silico interactions of natural and synthetic compounds with key proteins involved in Alzheimer’s disease: prospects for designing new therapeutics. Neurotox Res. 2023;41(5):408- 430. doi: 10.1007/s12640-023-00655-2

 

  1. Huang J, Jin Y, Chen Y, et al. Selected traditional Chinese medicine interventions for post-stroke cerebral edema: a review integrating clinical evidence and mechanistic insights. Front Pharmacol. 2025;16:1709821. doi: 10.3389/fphar.2025.1709821

 

  1. Lv F, Sun M, Qin C, et al. Study of the multitarget mechanism of Astragalus (HUANGQI) in the treatment of Alzheimer’s disease based on network pharmacology and molecular docking technology. Pharm Biol. 2024;62(1):634-647. doi: 10.1080/13880209.2024.2382962

 

  1. Mecca AP, van Dyck CH. Alzheimer’s & Dementia: The Journal of the Alzheimer’s Association. Alzheimers Dement. 2021;17(2):316-317. doi: 10.1002/alz.12190

 

  1. Chen C, Zhou Y, Wang J, et al. A modified CCK-8 assay for measuring cell viability in BV2 microglial cells. J Pharmacol Toxicol Methods. 2020;102:106674. doi: 10.1016/j.vascn.2020.106674

 

  1. Kim J, Kim DH, Kwak SG. Comprehensive guidelines for appropriate statistical analysis methods in research. Korean J Anesthesiol. 2024;77(5):503-517. doi: 10.4097/kja.24016

 

  1. Wang R, Sheng J, Li X. Piperine attenuates lipopolysaccharide-induced inflammatory responses in BV2 microglia through inhibition of NF-κB and MAPK signaling pathways. Neurochem Int. 2024;175:105718. doi: 10.1016/j.neuint.2024.105718

 

  1. Kwon Y, Kim J, Bae S, Lee C. Natural product-derived compounds as modulators of microglial inflammation in Alzheimer‘s disease: a systematic review. Int J Mol Sci. 2025;26(5):2104. doi: 10.3390/ijms26052104

 

  1. Kim M, Lee S, Cho K, Park H. Piperine attenuates lipopolysaccharide (LPS)-induced inflammatory responses in BV2 microglia. Int Immunopharmacol. 2017;42:209-218. doi: 10.1016/j.intimp.2016.11.001

 

  1. Alqahtani SM, Alqahtani AS, Alshehri S, et al. Pharmacological modulation of the PI3K/AKT/GSK3β axis: a new frontier in Alzheimer’s disease treatment. Inflammopharmacology. 2025;33(2):567-582. doi: 10.1007/s10787-025-02000-9

 

  1. Javed A, Youn K, Ho CT, et al. Unveiling the preventive potential of artesin in brain-heart axis: in-silico insights into Alzheimer’s and cardiovascular diseases prevention. Comput Biol Med. 2025;196(Pt C):110866. doi: 10.1016/j.compbiomed.2025.110866

 

  1. Gerschutz A, Heinsen H, Grunblatt E, et al. Neuron-specific alterations in signal transduction pathways associated with Alzheimer’s disease. J Alzheimers Dis. 2014;40(1):135-142. doi: 10.3233/JAD-131280

 

  1. Qian XH, Liu XL, Chen SD, et al. Integrating peripheral blood and brain transcriptomics to identify immunological features associated with Alzheimer’s disease in mild cognitive impairment patients. Front Immunol. 2022;13:986346. doi: 10.3389/fimmu.2022.986346

 

  1. Wei W, Yao JX, Zhang TT, et al. Network pharmacology reveals that berberine may function against Alzheimer’s disease via the AKT signaling pathway. Front Neurosci. 2023;17:1059496. doi: 10.3389/fnins.2023.1059496

 

  1. Tang L, Liu J, Xu X, et al. Pharmacological mechanism of Shen Huang Chong Ji for treating Alzheimer’s disease based on network pharmacology and experimental validation. Evid Based Complement Alternat Med. 2022;2022:9243348. doi: 10.1155/2022/9243348

 

  1. Zhang Y, Liu J, Yao M, Wang L. Flavonoids inhibiting tau hyperphosphorylation: implications for Alzheimer’s disease. Nat Prod Sci. 2025;31(2):75-83. doi: 10.20307/nps.2025.31.2.75

 

  1. Zhang T, Wei W, Chang S, et al. Integrated network pharmacology and comprehensive bioinformatics identifying the mechanisms and molecular targets of Yizhiqingxin formula for treatment of comorbidity with Alzheimer’s disease and depression. Front Pharmacol. 2022;13:853375. doi: 10.3389/fphar.2022.853375

 

  1. Wang D, Zheng J, Sun X, et al. Study on the pharmacological mechanism of icariin for the treatment of Alzheimer’s disease based on network pharmacology and molecular docking. Metabolites. 2023;14(1):10001. doi: 10.3390/metabo14010032

 

  1. Gnanarajan R, Uvarajan D, Ravichandran N, et al. Agnuside enhances neuronal survival and cognitive function by modulating PI3K/AKT/mTOR signaling in aluminium chloride induced Alzheimer’s disease. Prog Neuropsychopharmacol Biol Psychiatry. 2025;142:111532. doi: 10.1016/j.pnpbp.2025.111532

 

  1. Bolton CJ, Hohman TJ, Jefferson AL, et al. Increased neuroplastic activity in the pathogenesis of Alzheimer’s disease. Alzheimers Dement. 2025;21(11):e70897. doi: 10.1002/alz.70897

 

  1. Li M, Wang J, Chen L, Zhang Y. Targeting the PI3K/AKT signaling pathway in Alzheimer’s disease: from molecular mechanisms to therapeutic opportunities. Ageing Res Rev. 2025;95:102245. doi: 10.1016/j.arr.2025.102245

 

  1. Zhao H, Liu X, Chen S, et al. Salidroside alleviates Alzheimer’s disease-like pathology by activating PI3K/ AKT-mediated autophagy and anti-apoptosis. Phytother Res. 2025;39(2):789-803. doi: 10.1002/ptr.8345

 

  1. Chen MK, Mecca AP, Naganawa M, et al. Longitudinal assessment of synaptic density and cognitive decline in preclinical Alzheimer’s disease: a [11C]UCB-J PET cohort study. Alzheimers Dement. 2025;21(4):e14321. doi: 10.1002/alz.14321
Next article in this issue
Share
Back to top
Eurasian Journal of Medicine and Oncology, Electronic ISSN: 2587-196X Print ISSN: 2587-2400, Published by AccScience Publishing
We use cookies on our website to ensure you get the best experience.
Read more about our cookies here
Accept