Abstract
In this study, we aimed to evaluate the effects of the spirocyclopiperazinium salt compound LXM-15 on rheumatoid arthritis induced by complete Freund’s adjuvant (CFA) in rats and investigate the underlying mechanism. The results showed that LXM-15 significantly inhibited the paw edema and ankle swelling, and alleviated the mechanical allodynia and thermal hyperalgesia responses in the CFA rats. The histopathological results revealed that LXM-15 ameliorated the infiltration of inflammatory cells and joint destruction. The micro-CT scan showed that LXM-15 alleviated bone erosion and increased BMD in the ankle joints of the CFA rats. Western blot analyses showed that LXM-15 significantly reduced the upregulation of phospho-JAK2, phospho-STAT3, phospho-IκBα, and phospho-NF-κBp65, and the overexpression of BDNF in the dorsal root ganglions. ELISA result showed that the protein level of TNF-α in the paw tissue was decreased upon LXM-15 treatment. RT-PCR analysis showed that the mRNA expression levels of c-fos and BDNF were reduced in the dorsal root ganglions by LXM-15 treatment. The LXM-15-mediated anti-arthritic effects were abolished by treatment with hexamethonium (a peripheral nicotinic receptor antagonist), atropine methylnitrate (a peripheral muscarinic receptor antagonist), methyllycaconitine citrate (a selective α7 nicotinic receptor antagonist), and tropicamide (a selective M4 muscarinic receptor antagonist). Collectively, our results demonstrate that LXM-15 exerts anti-arthritic effects in CFA rats. The underlying mechanism may be related to the activation of the peripheral α7 nicotinic receptor and M4 muscarinic receptor by LXM-15, further suppressing the activation of the JAK2/STAT3 and IκBα/NF-κBp65 signaling pathways and, eventually, inhibiting the expression levels of TNF-α, BDNF, and c-fos.
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References
AlSharari, S.D., K. Freitas, and M.I. Damaj. 2013. Functional role of alpha7 nicotinic receptor in chronic neuropathic and inflammatory pain: studies in transgenic mice. Biochemical Pharmacology 86: 1201–1207.
De Jonge, W., and L. Ulloa. 2007. The alpha7 nicotinic acetylcholine receptor as a pharmacological target for inflammation. British Journal of Pharmacology 151: 915–929.
Egea, J., I. Buendia, E. Parada, E. Navarro, R. León, and M.G. Lopez. 2015. Anti-inflammatory role of microglial alpha7 nAChRs and its role in neuroprotection. Biochemical Pharmacology 97: 463–472.
El-Gaphar, O.A.M.A., A.M. Abo-Youssef, and G.K. Halal. 2018. Levetiracetam mitigates lipopolysaccharide-induced JAK2/STAT3 and TLR4/MAPK signaling pathways activation in a rat model of adjuvant-induced arthritis. European Journal of Pharmacology 826: 85–95.
El-Sayed, R., Y. Moustafa, and M. El-Azab. 2014. Evening primrose oil and celecoxib inhibited pathological angiogenesis, inflammation, and oxidative stress in adjuvant-induced arthritis: novel role of angiopoietin-1. Inflammopharmacology 22: 305–317.
Gao, X., Q. Sun, W. Zhang, Y. Jiang, R. Li, and J. Ye. 2018a. Anti-inflammatory effect and mechanism of the spirocyclopiperazinium salt compound LXM-15 in rats and mice. Inflammation Research 67: 363–370.
Gao, Y., H. Zhao, P. Wang, J. Wang, and L. Zou. 2018b. The roles of SOCS 3 and STAT 3 in bacterial infection and inflammatory diseases. Scandinavian Journal of Immunology 88: e12727.
Grimsholm, O., S. Rantapää-Dahlqvist, T. Dalén, and S. Forsgren. 2008. BDNF in RA: downregulated in plasma following anti-TNF treatment but no correlation with inflammatory parameters. Clinical Rheumatology 27: 1289–1297.
Helen, B., et al. 2015. Citrullinated peptide dendritic cell immunotherapy in HLA risk genotype-positive rheumatoid arthritis patients. Science Translational Medicine 7: 290ra287.
Lalisse, S., J. Hua, M. Lenoir, N. Linck, F. Rassendren, and L. Ulmann. 2018. Sensory neuronal P2RX4 receptors controls BDNF signaling in inflammatory pain. Scientific Reports 8: 964.
Li, R., L. Cai, D.-Y. Ren, X.-F. Xie, C.-M. Hu, and J. Li. 2012. Therapeutic effect of 7, 3′-dimethoxy hesperetin on adjuvant arthritis in rats through inhibiting JAK2-STAT3 signal pathway. International Immunopharmacology 14: 157–163.
Lin, Y.-T., L.-S. Ro, H.-L. Wang, and J.-C. Chen. 2011. Up-regulation of dorsal root ganglia BDNF and trkB receptor in inflammatory pain: an in vivo and in vitro study. Journal of Neuroinflammation 8: 126.
Liu, Y.-l., et al. 2009. Suppression of complete Freund’s adjuvant-induced adjuvant arthritis by cobratoxin. Acta Pharmacologica Sinica 30: 219.
Luo, J.G., et al. 2014. Activation of spinal NF-κB/p65 contributes to peripheral inflammation and hyperalgesia in rat adjuvant-induced arthritis. Arthritis & Rheumatology 66: 896–906.
Maanen, M.A.V., et al. 2009. The α7 nicotinic acetylcholine receptor on fibroblast-like synoviocytes and in synovial tissue from rheumatoid arthritis patients: a possible role for a key neurotransmitter in synovial inflammation. Arthritis and Rheumatism 60: 1272–1281.
Martino, G., et al. 2011. The M1/M4 preferring agonist xanomeline is analgesic in rodent models of chronic inflammatory and neuropathic pain via central site of action. Pain 152: 2852–2860.
McInnes, I.B., and G. Schett. 2011. The pathogenesis of rheumatoid arthritis. The New England Journal of Medicine 365: 2205–2219.
McNamee, K., R. Williams, and M. Seed. 2015. Animal models of rheumatoid arthritis: how informative are they? European Journal of Pharmacology 759: 278–286.
Mellado, M., L. Martínez-Muñoz, G. Cascio, P. Lucas, J.L. Pablos, and J.M. Rodríguez-Frade. 2015. T Cell migration in rheumatoid arthritis. Frontiers in Immunology 6: 384.
Moelants, E.A.V., M. Anneleen, V.D. Jo, and P. Paul. 2013. Regulation of TNF-α with a focus on rheumatoid arthritis. Immunology and Cell Biology 91: 393–401.
Morita, Y., N. Kashihara, M. Yamamura, H. Okamoto, S. Harada, M. Kawashima, and H. Makino. 1998. Antisense oligonucleotides targeting c-fos mRNA inhibit rheumatoid synovial fibroblast proliferation. Annals of the Rheumatic Diseases 57: 122–124.
Smolen, J.S., D. Aletaha, and I.B. Mcinnes. 2016. Rheumatoid arthritis. Lancet 388: 2023–2038.
Sullivan, N.R., et al. 2007. Pharmacological characterization of the muscarinic agonist (3R,4R)-3-(3-hexylsulfanyl-pyrazin-2-yloxy)-1-aza-bicyclo[2.2.1]heptane (WAY-132983) in in vitro and in vivo models of chronic pain. The Journal of Pharmacology and Experimental Therapeutics 322: 1294.
Voulgari, P.V. 2008. Emerging drugs for rheumatoid arthritis. Expert Opinion on Emerging Drugs 13: 175–196.
Wang, L., Z. Li, Y. Tan, Q. Li, H. Yang, P. Wang, J. Lu, and P. Liu. 2018. PARP1 interacts with STAT3 and retains active phosphorylated-STAT3 in nucleus during pathological myocardial hypertrophy. Molecular and Cellular Endocrinology 474: 137–150.
Yang, H., Q. Sun, Y. Liang, Y. Jiang, R. Li, and J. Ye. 2018. Antinociception of the spirocyclopiperazinium salt compound LXM-15 via activating α7 nAChR and M4 mAChR and inhibiting CaMKIIα/cAMP/CREB/CGRP signalling pathway in mice. Regulatory Toxicology and Pharmacology 94: 108–114.
Yoshida, K., T. Hashimoto, Y. Sakai, and A. Hashiramoto. 2014. Involvement of the circadian rhythm and inflammatory cytokines in the pathogenesis of rheumatoid arthritis. Journal of Immunology Research 2014: 282495.
Zhang, W., Q. Sun, X. Gao, Y. Jiang, R. Li, and J. Ye. 2013. Anti-inflammation of spirocyclopiperazinium salt compound LXM-10 targeting α7 nAChR and M4 mAChR and inhibiting JAK2/STAT3 pathway in rats. PLoS One 8: e66895.
Zhao, X., J. Ye, Q. Sun, Y. Xiong, R. Li, and Y. Jiang. 2011. Antinociceptive effect of spirocyclopiperazinium salt compound LXM-15 via activating peripheral α7 nAChR and M4 mAChR in mice. Neuropharmacology 60: 446–452.
Zimmermann, M. 1983. Ethical guidelines for investigations of experimental pain in conscious animals. Pain 16: 109–110.
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This research was supported by grants from the National Natural Science Foundation of People’s Republic of China (grant nos. 81870876 and 81470050).
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The animal-related protocols were approved by the Institutional Animal Care and Use Committee of Peking University (ethics approval number: LA2014189); the experiments were performed in accordance with the ethics guidelines of the International Association for the Study of Pain [30].
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Li, N., Liu, Q., Li, R.T. et al. Anti-arthritic Effect of the Spirocyclopiperazinium Salt Compound LXM-15 in Rats and Its Underlying Mechanism. Inflammation 43, 903–915 (2020). https://doi.org/10.1007/s10753-020-01177-1
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DOI: https://doi.org/10.1007/s10753-020-01177-1