P2X and P2Y Receptor Antagonists Reduce Inflammation in ATPinduced Microglia

Main Article Content

Amer Imraish https://orcid.org/0000-0003-1191-2905
Tuqa Abu-Thiab https://orcid.org/0000-0002-3054-4047
Hana Hammad https://orcid.org/0000-0003-3130-1749

Keywords

Microglia, P2X receptor, P2Y receptor, ATP, IL-1β, IL-6, TNF-α

Abstract

Background: P2 receptors have been implicated in the release of neurotransmitter and pro-inflammatory cytokines due to their response to neuroexcitatory substances in the microglia. The P2X4, P2X7 and P2Y12 receptors are involved in the development of pain behavior induced by peripheral nerve injury. However, it is not known if blocking P2X4, P2X7 and P2Y12 receptors is associated with the expression and the release of interleukin-1B (IL-1β), interleukin-6 (IL-6), or tumor necrosis factor-α (TNF-α) in cultured neonatal spinal cord microglia. Objective: For this reason, we examined the effects of P2X4, P2X7 and P2Y12 antagonists on the expression and the release of IL-1β, IL-6, and TNF-α in ATP-stimulated microglia. Methods: In this study, we observed the effect of A-740003, PSB-12062 and MRS 2395 (P2X4, P2X7 and P2Y12 receptors antagonist, respectively), on the expression and release of IL-1β, IL-6 and TNF-α by using real-time fluorescence quantitative polymerase chain reaction (PCR) and enzyme-linked immunosorbent assay (ELISA). Results: ATP induced the increased expression of IL-1β, IL-6 and TNF-α at the level of messenger RNA (mRNA). ATP-evoked increase in IL-1β, IL-6 and TNF-α mRNA expression was inhibited by the P2X4 receptor antagonist A-740003 or P2X7 receptor antagonist PSB-12062, respectively. Similarly, ATP-evoked release of IL-1β, IL-6 and TNF-α was inhibited by A-740003 and PSB-12062. Furthermore, ATP-evoked increased expression of Iba-1, IL-1β, IL-6 and TNF-α mRNA, and release of IL-1β, IL-6 and TNF-α were nearly all blocked after co-administration of A-740003 plus PSB-12062. Finally, ATP-evoked increased gene expression and release of IL-1β, IL-6 and TNF-α were also inhibited by MRS 2395 (P2Y12 antagonist). Conclusion: These observations suggest a new clue for therapeutic strategies to treat the neuro-inflammation.

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References

1. Eyo UB, Dailey ME. Microglia: key elements in neural development, plasticity, and pathology. J Neuroimmune Pharmacol. 2013;8(3):494-509. https://doi.org/10.1007/s11481-013-9434-z
2. Kaushal V, Schlichter LC. Mechanisms of microglia-mediated neurotoxicity in a new model of the stroke penumbra. J Neurosci. 2008;28(9):2221-2230. https://doi.org/10.1523/jneurosci.5643-07.2008
3. Smith JA, Das A, Ray SK, et al. Role of pro-inflammatory cytokines released from microglia in neurodegenerative diseases. Brain Res Bull. 2012;87(1):10-20. https://doi.org/10.1016/j.brainresbull.2011.10.004
4. Crain JM, Watters JJ. Microglial P2 Purinergic Receptor and Immunomodulatory Gene Transcripts Vary By Region, Sex, and Age in the Healthy Mouse CNS. Transcr Open Access. 2015;3(2). https://doi.org/10.4172/2329-8936.1000124
5. Burnstock G, Kennedy C. Is there a basis for distinguishing two types of P2-purinoceptor? General Pharmacology: The Vascular System. 1985;16(5):433-440. https://doi.org/10.1016/0306-3623(85)90001-1
6. Hide I, Tanaka M, Inoue A, et al. Extracellular ATP triggers tumor necrosis factor‐α release from rat microglia. Journal of Neurochemistry. 2000;75(3):965-972. https://doi.org/10.1046/j.1471-4159.2000.0750965.x
7. Coddou C, Yan Z, Obsil T, et al. Activation and regulation of purinergic P2X receptor channels. Pharmacological reviews. 2011;63(3):641-683. https://doi.org/10.1124/pr.110.003129
8. Tsuda M, Shigemoto-Mogami Y, Koizumi S, et al. P2X4 receptors induced in spinal microglia gate tactile allodynia after nerve injury. Nature. 2003;424(6950):778-783. https://doi.org/10.1038/nature01786
9. Zhang Z, Artelt M, Burnet M, et al. Lesional accumulation of P2X4 receptor+ monocytes following experimental traumatic brain injury. Experimental Neurology. 2006;197(1):252-257. https://doi.org/10.1016/j.expneurol.2005.09.015
10. Cavaliere F, Florenzano F, Amadio S, et al. Up-regulation of P2X2, P2X4 receptor and ischemic cell death: prevention by P2 antagonists. Neuroscience. 2003;120(1):85-98. https://doi.org/10.1016/s0306-4522(03)00228-8
11. Sandkuhler J. Models and mechanisms of hyperalgesia and allodynia. Physiological reviews. 2009;89(2):707-758. https://doi. org/10.1152/physrev.00025.2008
12. Labasi JM, Petrushova N, Donovan C, et al. Absence of the P2X7 receptor alters leukocyte function and attenuates an inflammatory response. The Journal of Immunology. 2002;168(12):6436-6445. https://doi.org/10.4049/jimmunol.168.12.6436
13. Quintas C, Pinho D, Pereira C, et al. Microglia P2Y6 receptors mediate nitric oxide release and astrocyte apoptosis. Journal of Neuroinflammation. 2014;11(1):1-12. https://doi.org/10.1186/s12974-014-0141-3
14. Morioka N, Tokuhara M, Harano S, et al. The activation of P2Y6 receptor in cultured spinal microglia induces the production of CCL2 through the MAP kinases-NF-κB pathway. Neuropharmacology. 2013;75:116-125. https://doi.org/10.1016/j.
neuropharm.2013.07.017
15. Tatsumi E, Yamanaka H, Kobayashi K, et al. RhoA/ROCK pathway mediates p38 MAPK activation and morphological changes downstream of P2Y12/13 receptors in spinal microglia in neuropathic pain. Glia. 2015;63(2):216-228. https://doi.org/10.1002/
glia.22745
16. Shigemoto‐Mogami Y, Koizumi S, Tsuda M, et al. Mechanisms underlying extracellular ATP‐evoked interleukin‐6 release in mouse microglial cell line, MG‐5. Journal of Neurochemistry. 2001;78(6):1339-1349. https://doi.org/10.1046/j.1471-
4159.2001.00514.x
17. Sung C-S, Wen Z-H, Chang W-K, et al. Intrathecal interleukin-1β administration induces thermal hyperalgesia by activating inducible nitric oxide synthase expression in the rat spinal cord. Brain Research. 2004;1015(1-2):145-153. https://doi.
org/10.1016/j.brainres.2004.04.068
18. Burnstock G, Fredholm B, North R, et al. The birth and postnatal development of purinergic signalling. Acta Physiologica. 2010;199(2):93-147. https://doi.org/10.1111/j.1748-1716.2010.02114.x
19. Idzko M, Ferrari D, Eltzschig HK. Nucleotide signalling during inflammation. Nature. 2014;509(7500):310-317. https://doi. org/10.1038/nature13085
20. Clark AK, Old EA, Malcangio M. Neuropathic pain and cytokines: current perspectives. Journal of Pain Research. 2013;6:803.https://doi.org/10.2147/jpr.s53660
21. Tsuda M, Masuda T, Tozaki-Saitoh H, et al. Microglial regulation of neuropathic pain. Journal of Pharmacological Sciences. 2013;121(2):89-94. https://doi.org/10.1254/jphs.12r14cp
22. Mitchell CH, Reigada D. Purinergic signalling in the subretinal space: a role in the communication between the retina and the RPE. Purinergic Signalling. 2008;4(2):101-107. https://doi.org/10.1007/s11302-007-9054-2
23. Fresta CG, Caruso G, Fidilio A, et al. Dihydrotanshinone, a natural diterpenoid, preserves blood-retinal barrier integrity via P2X7 receptor. International Journal of Molecular Sciences. 2020;21(23):9305. https://doi.org/10.3390/ijms21239305
24. Pérez-Flores G, Lévesque SA, Pacheco J, et al. The P2X7/P2X4 interaction shapes the purinergic response in murine macrophages. Biochemical and Biophysical Research Communications. 2015;467(3):484-490. https://doi.org/10.1016/j.bbrc.2015.10.025
25. Kobayashi K, Yamanaka H, Fukuoka T, et al. P2Y12 receptor upregulation in activated microglia is a gateway of p38 signaling and neuropathic pain. Journal of Neuroscience. 2008;28(11):2892-2902. https://doi.org/10.1523/jneurosci.5589-07.2008
26. Gu N, Eyo UB, Murugan M, et al. Microglial P2Y12 receptors regulate microglial activation and surveillance during neuropathic pain. Brain, behavior, and immunity. 2016;55:82-92. https://doi.org/10.1016/j.bbi.2015.11.007
27. Bekő K, Koványi B, Gölöncsér F, et al. Contribution of platelet P2Y12 receptors to chronic Complete Freund’s adjuvant‐induced inflammatory pain. Journal of Thrombosis and Haemostasis. 2017;15(6):1223-1235. https://doi.org/10.1111/jth.13684
28. Van Wageningen TA, Vlaar E, Kooij G, et al. Regulation of microglial TMEM119 and P2RY12 immunoreactivity in multiple sclerosis white and grey matter lesions is dependent on their inflammatory environment. Acta Neuropathologica Communications.
2019;7(1):1-16. https://doi.org/10.1186/s40478-019-0850-z
29. Maeda J, Minamihisamatsu T, Shimojo M, et al. Distinct microglial response against Alzheimer’s amyloid and tau pathologies characterized by P2Y12 receptor. Brain Communications. 2021;3(1):fcab011. https://doi.org/10.1093/braincomms/fcab011
30. Masuda T, Iwamoto S, Yoshinaga R, et al. Transcription factor IRF5 drives P2X4R+-reactive microglia gating neuropathic pain. Nature Communications. 2014;5(1):1-11. https://doi.org/10.3410/f.718384688.793495457
31. Krausgruber T, Blazek K, Smallie T, et al. IRF5 promotes inflammatory macrophage polarization and TH1-TH17 responses. Nature Immunology. 2011;12(3):231-238. https://doi.org/10.1038/ni.1990
32. Tsuda M, Toyomitsu E, Kometani M, et al. Mechanisms underlying fibronectin‐induced up‐regulation of P2X4R expression in microglia: distinct roles of PI3K–Akt and MEK–ERK signalling pathways. Journal of Cellular and Molecular Medicine.
2009;13(9b):3251-3259. https://doi.org/10.1111/j.1582-4934.2009.00719.x
33. Masuda T, Tsuda M, Yoshinaga R, et al. IRF8 is a critical transcription factor for transforming microglia into a reactive phenotype. Cell Reports. 2012;1(4):334-340. https://doi.org/10.1016/j.celrep.2012.02.014
34. Minten C, Terry R, Deffrasnes C, et al. IFN regulatory factor 8 is a key constitutive determinant of the morphological and molecular properties of microglia in the CNS. PloS one. 2012;7(11):e49851. https://doi.org/10.1371/journal.pone.0049851

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