Integrated in silico and in vitro analysis of oxyphyllacinol in low back pain–associated neuroinflammation

© 2026 by the Author(s). This article is an open access article distributed under the terms and conditions of the Creative Commons Attribution 4.0 International License ( https://creativecommons.org/licenses/by/4.0/ )

Download PDF

XML

Cite

Abstract

Neuroinflammation and sensitization-related mechanisms may contribute to persistent low back pain (LBP), motivating investigation of compounds that can modulate these processes. This study integrated network pharmacology, molecular docking, and in vitro experiments to investigate the anti-neuroinflammatory effects of oxyphyllacinol and its potential relevance to LBP-associated pathways. Candidate targets of oxyphyllacinol were predicted using the SwissTargetPrediction, BATMAN-TCM, and SuperPred databases, and the results were intersected with LBP-associated genes obtained from the GeneCards and OMIM databases. Protein–protein interaction network analysis identified PTGS2, TRPV1, CNR1, and COMT as candidate hub targets. Gene Ontology and Kyoto Encyclopedia of Genes and Genomes enrichment analyses indicated associations with neuroactive ligand–receptor interaction and with the regulation of TRP channels by inflammatory mediators. Molecular docking predicted favorable binding poses of oxyphyllacinol with the four candidate proteins. Biological validation was performed in lipopolysaccharide (LPS)-stimulated BV2 microglia. Oxyphyllacinol reduced LPS-stimulated PGE2, TNF-α, and IL-6 release, and decreased PTGS2 and TRPV1 messenger RNA expression, while increasing CNR1 and COMT messenger RNA expression (all p < 0.01). These findings demonstrate the anti-inflammatory activity of oxyphyllacinol in an in vitro microglial model and support further investigation of PTGS2-, TRPV1-, CNR1-, and COMT-associated pathways. Because the experimental validation was limited to an LPS-stimulated cell model, the results should be interpreted as preliminary mechanistic evidence rather than direct evidence of therapeutic efficacy in LBP.

Keywords

Oxyphyllacinol

Microglia

Neuroinflammation

Network pharmacology

Molecular docking

Low back pain

Funding

The work was funded by Guangxi University of Chinese Medicine (Grant No. 2024GXNSFAA297179).

Conflict of interest

The authors declare no competing interests.

References

Hartvigsen J, Hancock MJ, Kongsted A, et al. What low back pain is and why we need to pay attention. Lancet. 2018;391(10137):2356-2367. doi: 10.1016/S0140-6736(18)30480-X
GBD 2021 Low Back Pain Collaborators. Global, regional, and national burden of low back pain, 1990–2020, its attributable risk factors, and projections to 2050: a systematic analysis of the Global Burden of Disease Study 2021. Lancet Rheumatol. 2023;5(6):e316-e329. doi: 10.1016/S2665-9913(23)00098-X
Maher C, Underwood M, Buchbinder R. Non-specific low back pain. Lancet. 2017;389(10070):736-747. doi: 10.1016/S0140-6736(16)30970-9
Morris P, Ali K, Merritt M, Pelletier J, Macedo LG. A systematic review of the role of inflammatory biomarkers in acute, subacute and chronic non-specific low back pain. BMC Musculoskelet Disord. 2020;21(1):142. doi: 10.1186/s12891-020-3154-3
Licciardone JC, Kearns CM, Hodge LM, Bergamini MV. Associations of cytokine concentrations with key osteopathic lesions and clinical outcomes in patients with nonspecific chronic low back pain: results from the OSTEOPATHIC Trial. J Am Osteopath Assoc. 2012;112(9):596-605. doi: 10.7556/jaoa.2012.112.9.596
Ji RR, Nackley A, Huh Y, Terrando N, Maixner W. Neuroinflammation and Central Sensitization in Chronic and Widespread Pain. Anesthesiology. 2018;129(2):343-366. doi: 10.1097/ALN.0000000000002130
Inoue K, Tsuda M. Microglia in neuropathic pain: cellular and molecular mechanisms and therapeutic potential. Nat Rev Neurosci. 2018;19(3):138-152. doi: 10.1038/nrn.2018.2
Sessler K, Blechschmidt V, Hoheisel U, Mense S, Schirmer L, Treede RD. Spinal cord fractalkine (CX3CL1) signaling is critical for neuronal sensitization in experimental nonspecific, myofascial low back pain. J Neurophysiol. 2021;125(5):1598-1611. doi: 10.1152/jn.00348.2020
Foster NE, Anema JR, Cherkin D, et al. Prevention and treatment of low back pain: evidence, challenges, and promising directions. Lancet. 2018;391(10137):2368-2383. doi: 10.1016/S0140-6736(18)30489-6
Parisien M, Lima LV, Dagostino C, et al. Acute inflammatory response via neutrophil activation protects against the development of chronic pain. Sci Transl Med. 2022;14(644):eabj9954. doi: 10.1126/scitranslmed.abj9954
Qaseem A, Wilt TJ, McLean RM, Forciea MA. Noninvasive Treatments for Acute, Subacute, and Chronic Low Back Pain: A Clinical Practice Guideline From the American College of Physicians. Ann Intern Med. 2017;166(7):514-530. doi: 10.7326/M16-2367
Schreijenberg M, Koes BW, Lin CWC. Guideline recommendations on the pharmacological management of non-specific low back pain in primary care—is there a need to change? Expert Rev Clin Pharmacol. 2019;12(2):145-157. doi: 10.1080/17512433.2019.1565992
George SZ, Fritz JM, Silfies SP, et al. Interventions for the Management of Acute and Chronic Low Back Pain: Revision 2021. J Orthop Sports Phys Ther. 2021;51(11):CPG1-CPG60. doi: 10.2519/jospt.2021.0304
Zhai Y, Liu L, Zhang F, et al. Network pharmacology: a crucial approach in traditional Chinese medicine research. Chin Med. 2025;20:8. doi: 10.1186/s13020-024-01056-z
Hopkins AL. Network pharmacology: the next paradigm in drug discovery. Nat Chem Biol. 2008;4(11):682-690. doi: 10.1038/nchembio.118
Li S. Network pharmacology evaluation method guidance-draft. World J Tradit Chin Med. 2021;7(1):146-154. doi: 10.4103/wjtcm.wjtcm_11_21
Eberhardt J, Santos-Martins D, Tillack AF, Forli S. AutoDock Vina 1.2.0: New Docking Methods, Expanded Force Field, and Python Bindings. J Chem Inf Model. 2021;61(8):3891-3898. doi: 10.1021/acs.jcim.1c00203
Liao J, Zhao X. Recent Research Progress on the Chemical Constituents, Pharmacology, and Pharmacokinetics of Alpinae oxyphyllae Fructus. Molecules. 2024;29(16):3905. doi: 10.3390/molecules29163905
Qiu C, Mu L, Wang J, et al. Sesquiterpenoids from the fruits of Alpinia oxyphylla Miq. and their neuroprotective effect. Phytochemistry. 2023;211:113680. doi: 10.1016/j.phytochem.2023.113680
Shi W, Zhong J, Zhang Q, Yan C. Structural characterization and antineuroinflammatory activity of a novel heteropolysaccharide obtained from the fruits of Alpinia oxyphylla. Carbohydr Polym. 2020;229:115405. doi: 10.1016/j.carbpol.2019.115405
Dong J, Li H, Zhou M, Yao X, Geng J, Yu Y. Four new diarylheptanoids and two new terpenoids from the fruits of Alpinia oxyphylla and their anti-inflammatory activities. Chin J Nat Med. 2024;22(10):929-936. doi: 10.1016/S1875-5364(24)60723-5
Xu M, Yang Y, Peng J, et al. Effects of Alpinae Oxyphyllae Fructus on microglial polarization in a LPS-induced BV2 cells model of neuroinflammation via TREM2. J Ethnopharmacol. 2023;302:115914. doi: 10.1016/j.jep.2022.115914
Chang X, Zhang D, Shi W, et al. An arabinoxylan (AOP70-1) isolated from Alpinia oxyphylla alleviates neuroinflammation and neurotoxicity by TLR4/MyD88/NF-κB pathway. Int J Biol Macromol. 2024;277:134339. doi: 10.1016/j.ijbiomac.2024.134339
Wu B, Gan A, Wang R, Lin F, Yan T, Jia Y. Alpinia oxyphylla Miq. volatile oil ameliorates depressive behaviors and inhibits neuroinflammation in CUMS-exposed mice by inhibiting the TLR4-medicated MyD88/NF-κB signaling pathway. J Chem Neuroanat. 2023;130:102270. doi: 10.1016/j.jchemneu.2023.102270
Lin Y, Wongkrajang K, Shen X, et al. Discovery of diarylheptanoids that activate α7 nAchR-JAK2-STAT3 signaling in macrophages with anti-inflammatory activity in vitro and in vivo. Bioorg Med Chem. 2022;66:116811. doi: 10.1016/j.bmc.2022.116811
Mlost J, Kostrzewa M, Malek N, Starowicz K. Molecular Understanding of the Activation of CB1 and Blockade of TRPV1 Receptors: Implications for Novel Treatment Strategies in Osteoarthritis. Int J Mol Sci. 2018;19(2):342. doi: 10.3390/ijms19020342
Chen X, Chen Z, Ma G, et al. Reactive Oxygen Species Induced Upregulation of TRPV1 in Dorsal Root Ganglia Results in Low Back Pain in Rats. J Inflamm Res. 2024;17:2245-2256. doi: 10.2147/JIR.S446841
Mickle AD, Shepherd AJ, Mohapatra DP. Sensory TRP Channels: The Key Transducers of Nociception and Pain. In: Progress in Molecular Biology and Translational Science. Elsevier; 2015;131:73-118. doi: 10.1016/bs.pmbts.2015.01.002
Starowicz K, Di Marzo V. Non-psychotropic analgesic drugs from the endocannabinoid system: “magic bullet” or “multiple-target” strategies? Eur J Pharmacol. 2013;716(1-3):41-53. doi: 10.1016/j.ejphar.2013.01.075
Vetterlein A, Monzel M, Reuter M. Are catechol-O-methyltransferase gene polymorphisms genetic markers for pain sensitivity after all? A review and meta-analysis. Neurosci Biobehav Rev. 2023;148:105112. doi: 10.1016/j.neubiorev.2023.105112
Li X, Wei S, Niu S, et al. Network pharmacology prediction and molecular docking-based strategy to explore the potential mechanism of Huanglian Jiedu Decoction against sepsis. Comput Biol Med. 2022;144:105389. doi: 10.1016/j.compbiomed.2022.105389
Gfeller D, Grosdidier A, Wirth M, Daina A, Michielin O, Zoete V. SwissTargetPrediction: a web server for target prediction of bioactive small molecules. Nucleic Acids Res. 2014;42(W1):W32-W38. doi: 10.1093/nar/gku293
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
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
Laskowski RA, Swindells MB. LigPlot+: multiple ligand-protein interaction diagrams for drug discovery. J Chem Inf Model. 2011;51(10):2778-2786. doi: 10.1021/ci200227u
Huang P, Zhang Z, Zhang P, et al. TREM2 deficiency aggravates NLRP3 inflammasome activation and pyroptosis in MPTP-induced Parkinson’s disease mice and LPS-induced BV2 cells. Mol Neurobiol. 2024;61(5):2590-2605. doi: 10.1007/s12035-023-03713-0
Jin CY, Lee JD, Park C, Choi YH, Kim GY. Curcumin attenuates the release of pro-inflammatory cytokines in lipopolysaccharide-stimulated BV2 microglia. Acta Pharmacol Sin. 2007;28(10):1645-1651. doi: 10.1111/j.1745-7254.2007.00651.x
Bao Y, Zhu Y, He G, et al. Dexmedetomidine attenuates neuroinflammation in LPS-stimulated BV2 microglia cells through upregulation of miR-340. Drug Des Devel Ther. 2019;13:3465-3475. doi: 10.2147/DDDT.S210511
Munoz OB. Determination of Proinflammatory and Antiinflammatory Cytokines by ELISA Technique. In: Matson RS, eds. ELISA. Humana; 2023:101-108. Walker JM, ed. Methods in Molecular Biology; vol 2612. doi: 10.1007/978-1-0716-2903-1_8
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2^−ΔΔCT method. Methods. 2001;25(4):402-408. doi: 10.1006/meth.2001.1262
Spinsanti G, Zannolli R, Panti C, et al. Quantitative Real-Time PCR detection of TRPV1-4 gene expression in human leukocytes from healthy and hyposensitive subjects. Mol Pain. 2008;4:51. doi: 10.1186/1744-8069-4-51
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

Previous article in this issue

Next article in this issue

Advanced Neurology, Electronic ISSN: 2810-9619 Print ISSN: 3060-8589, Published by AccScience Publishing