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Interleukin-4 signalling pathway underlies the anxiolytic effect induced by 3-deoxyadenosine

  • Tangxin Gao
  • Bai Li
  • Yangyang Hou
  • Shaolei Luo
  • Lei Feng
  • Jun Nie
  • Yi Ma
  • Le Xiao
  • Xu Chen
  • Hongkun Bao
  • Xianmin Lu
  • Feilong Huang
  • Gang Wang
  • Chunjie Xiao
  • Jing DuEmail author
Original Investigation
  • 51 Downloads

Abstract

Rationale

Converging evidence suggests that neuroimmunity plays an important role in the pathophysiology of anxiety. Interleukin (IL)-4 is a key cytokine regulating neuroimmune functions in the central nervous system. More efficient anxiolytics with neuro-immune mechanisms are urgently needed.

Objective

To determine whether 3′-deoxyadenosine (3′-dA) exerts an anxiolytic effect and to examine the role of IL-4 in the anxiolytic effect of 3′-dA in mice.

Methods

We investigated the effects of 3′-dA on anxiety-like behaviors using elevated plus maze (EPM) or light-dark box (LDB) tests after 45 min or 5 days of treatment. Expression of IL-4, IL-10, IL-1β, TNF-α, and IL-6 in the prefrontal cortex (PFC) was detected by Western blot and/or double immunostaining. Intracerebroventricular injection of RIL-4Rα (an IL-4-specific inhibitor) and intraperitoneal injection of 3′-dA or imipramine were co-administered, followed by EPM test.

Results

3′-dA exhibited a stronger and faster anxiolytic effect than imipramine in behavioral tests. Furthermore, 3′-dA enhanced IL-4 expression after 45 min or 5 days, TNF-α and IL-1β expression decreased significantly after a 5-day treatment with 3′-dA, and IL-10 expression increased after a 5-day treatment with 3′-dA or imipramine in the PFC. IL-4 was expressed in neurons and in some astrocytes and microglia. IL-4 expression showed a strong positive correlation with reduced anxiety behaviors. RIL-4Rα completely blocked the anxiolytic effects induced by 3′-dA and imipramine.

Conclusions

This study identifies a novel and common anxiolytic IL-4 signaling pathway and provides an innovative drug with a novel neuro-immune mechanism for treating anxiety disorder.

Keywords

3′-deoxyadenosine Anxiolytic effect Neuroimmune Cytokines Interleukin-4 

Notes

Funding information

This work was supported by the National Natural Science Foundation of China (Grant No. 31560274, 31650005, and 81760651), Yunnan Province Funding (Grant No. KC1710123), and the Yunnan High-level Professional Funding (Grant No. 2012HA004) from the Department of Science and Technology of Yunnan Province.

Compliance with ethical standards

Conflict of interests

The authors declare that they have no conflict of interest.

Ethical approval

All experimental procedures were performed in accordance with the Medical Ethics Committee of the School of Medicine of Yunnan University and followed the Guide for the Care and Use of Laboratory Animals (ISBN 0-309-05377-3).

References

  1. Adhikari A, Topiwala MA, Gordon JA (2010) Synchronized activity between the ventral hippocampus and the medial prefrontal cortex during anxiety. Neuron 65:257–269CrossRefGoogle Scholar
  2. Adhikari A, Topiwala MA, Gordon JA (2011) Single units in the medial prefrontal cortex with anxiety-related firing patterns are preferentially influenced by ventral hippocampal activity. Neuron 71:898–910CrossRefGoogle Scholar
  3. Barnes PJ (2011) Glucocorticosteroids: current and future directions. Br J Pharmacol 163:29–43CrossRefGoogle Scholar
  4. Birn RM, Shackman AJ, Oler JA, Williams LE, McFarlin DR, Rogers GM, Shelton SE, Alexander AL, Pine DS, Slattery MJ, Davidson RJ, Fox AS, Kalin NH (2014) Evolutionarily conserved prefrontal-amygdalar dysfunction in early-life anxiety. Mol Psychiatry 19:915–922CrossRefGoogle Scholar
  5. Caddy C, Giaroli G, White TP, Shergill SS, Tracy DK (2014) Ketamine as the prototype glutamatergic antidepressant: pharmacodynamic actions, and a systematic review and meta-analysis of efficacy. Ther Adv Psychopharmacol 4:75–99CrossRefGoogle Scholar
  6. Clarke RM, Lyons A, O'Connell F, Deighan BF, Barry CE, Anyakoha NG, Nicolaou A, Lynch MA (2008) A pivotal role for interleukin-4 in atorvastatin-associated neuroprotection in rat brain. J Biol Chem 283:1808–1817CrossRefGoogle Scholar
  7. Cox BJ, Endler NS, Lee PS, Swinson RP (1992) A meta-analysis of treatments for panic disorder with agoraphobia: imipramine, alprazolam, and in vivo exposure. J Behav Ther Exp Psychiatry 23:175–182CrossRefGoogle Scholar
  8. Craske MG, Stein MB (2016) Anxiety. Lancet 388:3048–3059CrossRefGoogle Scholar
  9. Curtin NM, Mills KH, Connor TJ (2009) Psychological stress increases expression of IL-10 and its homolog IL-19 via beta-adrenoceptor activation: reversal by the anxiolytic chlordiazepoxide. Brain Behav Immun 23:371–379CrossRefGoogle Scholar
  10. Diogo GR, Sparrow A, Paul MJ, Copland A, Hart PJ, Stelter S, van Dolleweerd C, Drake PMW, Macallan DC, Reljic R (2017) Murine IL-4Delta2 splice variant down-regulates IL-4 activities independently of IL-4Ralpha binding and STAT-6 phosphorylation. Cytokine 99:154–162CrossRefGoogle Scholar
  11. Dorn LD, Gayles JG, Engeland CG, Houts R, Cizza G, Denson LA (2016) Cytokine patterns in healthy adolescent girls: heterogeneity captured by variable and person-centered statistical strategies. Psychosom Med 78:646–656CrossRefGoogle Scholar
  12. Frye CA (2014) Endocrine-disrupting chemicals: elucidating our understanding of their role in sex and gender-relevant end points Vitamins & Hormones, pp Volume 94, 2014, Pages 41–98Google Scholar
  13. Gomes EP, Aguiar JC, Fonseca-Silva T, Dias LC, Moura-Boas KP, Roy A, Velloso NA, Rodrigues-Neto JF, De-Paula AM, Guimaraes AL (2013) Diazepam reverses the alveolar bone loss and hippocampal interleukin-1beta and interleukin-6 enhanced by conditioned fear stress in ligature-induced periodontal disease in rats. J Periodontal Res 48:151–158CrossRefGoogle Scholar
  14. Gommoll C, Durgam S, Mathews M, Forero G, Nunez R, Tang X, Thase ME (2015) A double-blind, randomized, placebo-controlled, fixed-dose phase III study of vilazodone in patients with generalized anxiety disorder. Depress Anxiety 32:451–459CrossRefGoogle Scholar
  15. Han A, Yeo H, Park MJ, Kim SH, Choi HJ, Hong CW, Kwon MS (2015) IL-4/10 prevents stress vulnerability following imipramine discontinuation. J Neuroinflammation 12:197CrossRefGoogle Scholar
  16. Hirano S (2012) Western blot analysis. Methods Mol Biol 926:87–97CrossRefGoogle Scholar
  17. Hu Z, Lee CI, Shah VK, Oh EH, Han JY, Bae JR, Lee K, Chong MS, Hong JT, Oh KW (2013) Cordycepin increases nonrapid eye movement sleep via adenosine receptors in rats. Evid Based Complement Alternat Med 2013:840134PubMedPubMedCentralGoogle Scholar
  18. Huang H, Zhang X, Fu X, Zhang X, Lang B, Xiang X, Hao W (2018) Alcohol-induced conditioned place preference negatively correlates with anxiety-like behavior in adolescent mice: inhibition by a neurokinin-1 receptor antagonist. Psychopharmacology 235:2847–2857CrossRefGoogle Scholar
  19. Hwang J, Zheng LT, Ock J, Lee MG, Kim SH, Lee HW, Lee WH, Park HC, Suk K (2008) Inhibition of glial inflammatory activation and neurotoxicity by tricyclic antidepressants. Neuropharmacology 55:826–834CrossRefGoogle Scholar
  20. Karlsson L, Nousiainen N, Scheinin NM, Maksimow M, Salmi M, Lehto SM, Tolvanen M, Lukkarinen H, Karlsson H (2017) Cytokine profile and maternal depression and anxiety symptoms in mid-pregnancy-the FinnBrain Birth Cohort Study. Arch Womens Ment Health 20:39–48CrossRefGoogle Scholar
  21. Kokare DM, Shelkar GP, Borkar CD, Nakhate KT, Subhedar NK (2011) A simple and inexpensive method to fabricate a cannula system for intracranial injections in rats and mice. J Pharmacol Toxicol Methods 64:246–250CrossRefGoogle Scholar
  22. Lee HJ, Park HJ, Starkweather A, An K, Shim I (2016) Decreased Interleukin-4 release from the neurons of the locus coeruleus in response to immobilization stress. Mediat Inflamm 2016:3501905Google Scholar
  23. Levitan MN, Papelbaum M, Nardi AE (2015) Profile of agomelatine and its potential in the treatment of generalized anxiety disorder. Neuropsychiatr Dis Treat 11:1149–1155CrossRefGoogle Scholar
  24. Li JY, Boado RJ, Pardridge WM (2001) Cloned blood-brain barrier adenosine transporter is identical to the rat concentrative Na+ nucleoside cotransporter CNT2. J Cereb Blood Flow Metab 21:929–936CrossRefGoogle Scholar
  25. Li Z, Zhang Z, Ming WK, Chen X, Xiao XM (2017) Tracing GFP-labeled WJMSCs in vivo using a chronic salpingitis model: an animal experiment. Stem Cell Res Ther 8:272CrossRefGoogle Scholar
  26. Likhtik E, Stujenske JM, Topiwala MA, Harris AZ, Gordon JA (2014) Prefrontal entrainment of amygdala activity signals safety in learned fear and innate anxiety. Nat Neurosci 17:106–113CrossRefGoogle Scholar
  27. Lucas EK, Wu WC, Roman-Ortiz C, Clem RL (2018) Prazosin during fear conditioning facilitates subsequent extinction in male C57Bl/6N mice. Psychopharmacology.  https://doi.org/10.1007/s00213-018-5001-x CrossRefGoogle Scholar
  28. Lyons A, Griffin RJ, Costelloe CE, Clarke RM, Lynch MA (2007) IL-4 attenuates the neuroinflammation induced by amyloid-beta in vivo and in vitro. J Neurochem 101(3):771–781.  https://doi.org/10.1111/j.1471-4159.2006.04370.x CrossRefPubMedGoogle Scholar
  29. Lyons A, McQuillan K, Deighan BF, O'Reilly JA, Downer EJ, Murphy AC, Watson M, Piazza A, O'Connell F, Griffin R, Mills KH, Lynch MA (2009) Decreased neuronal CD200 expression in IL-4-deficient mice results in increased neuroinflammation in response to lipopolysaccharide. Brain Behav Immun 23:1020–1027CrossRefGoogle Scholar
  30. Maher FO, Nolan Y, Lynch MA (2005) Downregulation of IL-4-induced signalling in hippocampus contributes to deficits in LTP in the aged rat. Neurobiol Aging 26:717–728CrossRefGoogle Scholar
  31. Mello NK, Mendelson JH (2009) Cocaine, hormones, and behavior: clinical and preclinical studies. Horm Brain Behav (second edition), pp part V, 2009, Pages 3081–3140CrossRefGoogle Scholar
  32. Moon ML, Joesting JJ, Blevins NA, Lawson MA, Gainey SJ, Towers AE, McNeil LK, Freund GG (2015) IL-4 knock out mice display anxiety-like behavior. Behav Genet 45:451–460CrossRefGoogle Scholar
  33. Nolan Y, Maher FO, Martin DS, Clarke RM, Brady MT, Bolton AE, Mills KH, Lynch MA (2005) Role of interleukin-4 in regulation of age-related inflammatory changes in the hippocampus. J Biol Chem 280:9354–9362CrossRefGoogle Scholar
  34. Obuchowicz E, Bielecka AM, Paul-Samojedny M, Pudelko A, Kowalski J (2014) Imipramine and fluoxetine inhibit LPS-induced activation and affect morphology of microglial cells in the rat glial culture. Pharmacol Rep: PR 66:34–43CrossRefGoogle Scholar
  35. Pardridge WM (2012) Drug transport across the blood-brain barrier. J Cereb Blood Flow Metab 32:1959–1972CrossRefGoogle Scholar
  36. Quagliato LA, Nardi AE (2017) Cytokine alterations in panic disorder: a systematic review. J Affect Disord 228:91–96CrossRefGoogle Scholar
  37. Ramirez K, Sheridan JF (2016) Antidepressant imipramine diminishes stress-induced inflammation in the periphery and central nervous system and related anxiety- and depressive- like behaviors. Brain Behav Immun 57:293–303CrossRefGoogle Scholar
  38. Ramirez K, Shea DT, McKim DB, Reader BF, Sheridan JF (2015) Imipramine attenuates neuroinflammatory signaling and reverses stress-induced social avoidance. Brain Behav Immun 46:212–220CrossRefGoogle Scholar
  39. Rao YP, Sugasini D, Lokesh BR (2016) Dietary gamma oryzanol plays a significant role in the anti-inflammatory activity of rice bran oil by decreasing pro-inflammatory mediators secreted by peritoneal macrophages of rats. Biochem Biophys Res Commun 479:747–752CrossRefGoogle Scholar
  40. Seo MJ, Kim MJ, Lee HH, Park JU, Kang BW, Kim GY, Rhu EJ, Kim JI, Kim KH, Jeong YK (2013) Effect of cordycepin on the expression of the inflammatory cytokines TNF-alpha, IL-6, and IL-17A in C57BL/6 mice. J Microbiol Biotechnol 23:156–160CrossRefGoogle Scholar
  41. Shin S, Moon S, Park Y, Kwon J, Lee S, Lee CK, Cho K, Ha NJ, Kim K (2009) Role of cordycepin and adenosine on the phenotypic switch of macrophages via induced anti-inflammatory cytokines. Immune Netw 9:255–264CrossRefGoogle Scholar
  42. Shukla R, Prevot TD, French L, Isserlin R, Rocco BR, Banasr M, Bader GD, Sibille E (2019) The relative contributions of cell-dependent cortical microcircuit aging to cognition and anxiety. Biol Psychiatry 85:257–267CrossRefGoogle Scholar
  43. Stein MB, Stein DJ (2008) Social anxiety disorder. Lancet 371:1115–1125CrossRefGoogle Scholar
  44. Tovote P, Fadok JP, Luthi A (2015) Neuronal circuits for fear and anxiety. Nat Rev Neurosci 16:317–331CrossRefGoogle Scholar
  45. Tuli HS, Sharma AK, Sandhu SS, Kashyap D (2013) Cordycepin: a bioactive metabolite with therapeutic potential. Life Sci 93:863–869CrossRefGoogle Scholar
  46. Venmar KT, Carter KJ, Hwang DG, Dozier EA, Fingleton B (2014) IL4 receptor ILR4alpha regulates metastatic colonization by mammary tumors through multiple signaling pathways. Cancer Res 74:4329–4340CrossRefGoogle Scholar
  47. Vogelzangs N, Beekman AT, de Jonge P, Penninx BW (2013) Anxiety disorders and inflammation in a large adult cohort. Transl Psychiatry 3:e249CrossRefGoogle Scholar
  48. Zhao X, Wang H, Sun G, Zhang J, Edwards NJ, Aronowski J (2015) Neuronal Interleukin-4 as a modulator of microglial pathways and ischemic brain damage. J Neurosci 35:11281–11291CrossRefGoogle Scholar
  49. Zhou X, Meyer CU, Schmidtke P, Zepp F (2002) Effect of cordycepin on interleukin-10 production of human peripheral blood mononuclear cells. Eur J Pharmacol 453:309–317CrossRefGoogle Scholar

Copyright information

© Springer-Verlag GmbH Germany, part of Springer Nature 2019

Authors and Affiliations

  • Tangxin Gao
    • 1
    • 2
  • Bai Li
    • 1
    • 2
  • Yangyang Hou
    • 1
    • 2
  • Shaolei Luo
    • 1
    • 2
  • Lei Feng
    • 3
  • Jun Nie
    • 1
    • 2
  • Yi Ma
    • 1
    • 2
  • Le Xiao
    • 3
  • Xu Chen
    • 3
  • Hongkun Bao
    • 1
    • 2
  • Xianmin Lu
    • 4
  • Feilong Huang
    • 5
  • Gang Wang
    • 3
  • Chunjie Xiao
    • 1
    • 2
  • Jing Du
    • 1
    • 2
    • 4
    Email author
  1. 1.State Key Laboratory for Conservation and Utilization of Bio-resources in YunnanYunnan UniversityKunmingChina
  2. 2.School of MedicineYunnan UniversityKunmingChina
  3. 3.The National Clinical Research Center for Mental Disorders & Beijing Key Laboratory of Mental Disorders & Beijing Anding HospitalCapital Medical UniversityBeijingChina
  4. 4.Beijing Gragen Biotechnology Co. LtdBeijingChina
  5. 5.Beijing Los-Portland Academy of MedicineBeijingChina

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