Abstract
Anxiety disorders and cognitive decline are highly prevalent in rheumatic diseases, including Juvenile Idiopathic Arthritis (JIA). In this study, we investigated the effect of long-term treatment with infliximab and tocilizumab on anxiety-like behaviour and cognitive performance in a juvenile collagen–induced arthritis (CIA) rat model. Forty-nine rats with established moderate arthritis were randomly allocated into 7 equal groups: negative control, vehicle, methotrexate, infliximab, tocilizumab, methotrexate + infliximab and methotrexate + tocilizumab groups. Behavioural tests were performed to evaluate anxiety-like behaviour and cognitive function. Neuropathological changes were investigated by histological examination at the level of the hippocampus, the amygdala and the prefrontal cortex. Also, the expression of Brain-Derived Neurotrophic Factor (BDNF), a biomarker associated with neuronal survival and plasticity, was determined in the hippocampus and the amygdala by RT-qPCR. We found that both infliximab and tocilizumab reduced anxiety-like behaviour in the elevated-plus and elevated-zero maze tests. Tocilizumab, also, improved cognitive function in the olfactory social memory and passive avoidance tests. Anti-cytokine treatment reversed the histopathological changes in the brain induced by CIA. BDNF expression was higher in all treatment groups and especially those receiving monoclonal antibodies combined with methotrexate. Our data provide evidence that chronic infliximab and tocilizumab treatment reduces anxiety-like behaviour, improves cognitive function, reverses neuropathological changes and increases central BDNF expression in a juvenile arthritis rat model. These findings may be translated to humans to address behavioural comorbidities associated with JIA.
Similar content being viewed by others
AVAILABILITY OF DATA AND MATERIAL
The data underlying this article will be shared on reasonable request to the corresponding author.
CODE AVAILABILITY
Not applicable.
References
Ravelli, A., and A. Martini. 2007. Juvenile idiopathic arthritis. Lancet 369: 767–778.
Thierry, S., B. Fautrel, I. Lemelle, and F. Guillemin. 2014. Prevalence and incidence of juvenile idiopathic arthritis: A systematic review. Joint, Bone, Spine 81: 112–117.
Giancane, G., A. Alongi, and A. Ravelli. 2017. Update on the pathogenesis and treatment of juvenile idiopathic arthritis. Current Opinion in Rheumatology 29: 523–529.
Appenzeller, S., M.B. Bertolo, and L.T. Costallat. 2004. Cognitive impairment in rheumatoid arthritis. Methods and Findings in Experimental and Clinical Pharmacology 26: 339–343.
Fiest, K.M., C.A. Hitchon, C.N. Bernstein, C.A. Peschken, J.R. Walker, L.A. Graff, R. Zarychanski, A. Abou-Setta, S.B. Patten, J. Sareen, J. Bolton, R.A. Marrie, and the CIHR team. . 2017. Systematic review and meta-analysis of interventions for depression and anxiety in persons with rheumatoid arthritis. Journal of Clinical Rheumatology 23: 425–434.
Meade, T., N. Manolios, S.R. Cumming, P.G. Conaghan, and P. Katz. 2018. Cognitive impairment in rheumatoid arthritis: A systematic review. Arthritis Care Res (Hoboken) 70: 39–52.
Fair, D.C., M. Rodriguez, A.M. Knight, and T.B. Rubinstein. 2019. Depression and anxiety in patients with juvenile idiopathic arthritis: Current insights and impact on quality of life, a systematic review. Open Access Rheumatology: Research and Reviews 11: 237–252.
Machin, A.R., O. Babatunde, R. Haththotuwa, I. Scott, M. Blagojevic-Bucknall, N. Corp, C.A. Chew-Graham, and S.L. Hider. 2020. The association between anxiety and disease activity and quality of life in rheumatoid arthritis: A systematic review and meta-analysis. Clinical Rheumatology 39: 1471–1482.
Quan, N. 2008. Immune-to-brain signaling: How important are the blood-brain barrier-independent pathways? Molecular Neurobiology 37: 142–152.
Chavan, S.S., V.A. Pavlov, and K.J. Tracey. 2017. Mechanisms and therapeutic relevance of neuro-immune communication. Immunity 46: 927–942.
Dantzer, R. 2018. Neuroimmune interactions: From the brain to the immune system and vice versa. Physiological Reviews 98: 477–504.
Banks, W.A., A.J. Kastin, and R.D. Broadwell. 1995. Passage of cytokines across the blood-brain barrier. NeuroImmunoModulation 2: 241–248.
Galic, M.A., K. Riazi, and Q.J. Pittman. 2012. Cytokines and brain excitability. Frontiers in Neuroendocrinology 33: 116–125.
Loftis, J.M., M. Huckans, and B.J. Morasco. 2010. Neuroimmune mechanisms of cytokine-induced depression: Current theories and novel treatment strategies. Neurobiology of Diseases 37: 519–533.
Himmerich, H., O. Patsalos, N. Lichtblau, M.A.A. Ibrahim, and B. Dalton. 2019. Cytokine research in depression: Principles, challenges, and open questions. Front Psychiatry. 10: 30.
Chandrashekara, S., K. Jayashree, H.B. Veeranna, H.S. Vadiraj, M.N. Ramesh, A. Shobha, Y. Sarvanan, and Y.K. Vikram. 2007. Effects of anxiety on TNF-alpha levels during psychological stress. Journal of Psychosomatic Research 63: 65–69.
Duivis, H.E., N. Vogelzangs, N. Kupper, P. de Jonge, and B.W. Penninx. 2013. Differential association of somatic and cognitive symptoms of depression and anxiety with inflammation: Findings from the Netherlands Study of Depression and Anxiety (NESDA). Psychoneuroendocrinology 38: 1573–1585.
Alshogran, O.Y., A.A. Khalil, A.O. Oweis, S.M. Altawalbeh, and M.A.Y. Alqudah. 2018. Association of brain-derived neurotrophic factor and interleukin-6 serum levels with depressive and anxiety symptoms in hemodialysis patients. General Hospital Psychiatry 53: 25–31.
Trapero, I., and O. Cauli. 2014. Interleukin 6 and cognitive dysfunction. Metabolic Brain Disease 29: 593–608.
Rosenblat, J.D., E. Brietzke, R.B. Mansur, N.A. Maruschak, Y. Lee, and R.S. McIntyre. 2015. Inflammation as a neurobiological substrate of cognitive impairment in bipolar disorder: Evidence, pathophysiology and treatment implications. Journal of Affective Disorders 188: 149–159.
Kim, Y.S., K.J. Lee, and H. Kim. 2017. Serum tumour necrosis factor-alpha and interleukin-6 levels in Alzheimer’s disease and mild cognitive impairment. Psychogeriatrics 17: 224–230.
Shen, X.N., L.D. Niu, Y.J. Wang, X.P. Cao, Q. Liu, L. Tan, C. Zhang, and J.T. Yu. 2019. Inflammatory markers in Alzheimer’s disease and mild cognitive impairment: A meta-analysis and systematic review of 170 studies. Journal of Neurology, Neurosurgery and Psychiatry 90: 590–598.
Bathina, S., and U.N. Das. 2015. Brain-derived neurotrophic factor and its clinical implications. Archives of Medical Science 11: 1164–1178.
Felger, J.C., and F.E. Lotrich. 2013. Inflammatory cytokines in depression: Neurobiological mechanisms and therapeutic implications. Neuroscience 246: 199–229.
Suliman, S., S.M. Hemmings, and S. Seedat. 2013. Brain-Derived Neurotrophic Factor (BDNF) protein levels in anxiety disorders: Systematic review and meta-regression analysis. Frontiers in Integrative Neuroscience 7: 55.
Kilkenny, C., Browne, W. J., Cuthill, I. C., Emerson, M., Altman, D. G. 2010. Improving bioscience research reporting: the ARRIVE guidelines for reporting animal research. PLoS Biol. 8:e1000412.
Rosloniec, E. F., Cremer, M., Kang, A. H., Myers, L. K., Brand, D. D. 2010. Collagen-induced arthritis. Curr Protoc Immunol Chapter 15: Unit 15 5 1–25.
Bilasy, S.E., S.S. Essawy, M.F. Mandour, E.A. Ali, and S.A. Zaitone. 2015. Myelosuppressive and hepatotoxic potential of leflunomide and methotrexate combination in a rat model of rheumatoid arthritis. Pharmacological Reports 67: 102–114.
Karson, A., T. Demirtas, D. Bayramgurler, F. Balci, and T. Utkan. 2013. Chronic administration of infliximab (TNF-alpha inhibitor) decreases depression and anxiety-like behaviour in rat model of chronic mild stress. Basic & Clinical Pharmacology & Toxicology 112: 335–340.
Taskin, M.I., A.C. Gungor, E. Adali, A. Yay, G.O. Onder, and U. Inceboz. 2016. A humanized anti-interleukin 6 receptor monoclonal antibody, tocilizumab, for the treatment of endometriosis in a rat model. Reproductive Sciences 23: 662–669.
Marques, M.R., F. Stigger, E. Segabinazi, O.A. Augustin, S. Barbosa, F.V. Piazza, M. Achaval, and S. Marcuzzo. 2014. Beneficial effects of early environmental enrichment on motor development and spinal cord plasticity in a rat model of cerebral palsy. Behavioural Brain Research 263: 149–157.
Rosique, C., D. Lebsir, P. Lestaevel, S. Benatia, P. Guigon, F. Caire-Maurisier, M. Benderitter, D. Bennouna, M. Souidi, and J.C. Martin. 2019. Assessment of the effects of repeated doses of potassium iodide intake during pregnancy on male and female rat offspring using metabolomics and lipidomics. Journal of Toxicology and Environmental Health. Part A 82: 603–615.
Kouvelas, D., C. Pourzitaki, G. Papazisis, K. Tsilkos, M. Chourdakis, and M.M. Kraus. 2009. Chronic aortic denervation decreases anxiety and impairs social memory in rats. Life Sciences 85: 602–608.
Walf, A.A., and C.A. Frye. 2007. The use of the elevated plus maze as an assay of anxiety-related behavior in rodents. Nature Protocols 2: 322–328.
Shepherd, J.K., S.S. Grewal, A. Fletcher, D.J. Bill, and C.T. Dourish. 1994. Behavioural and pharmacological characterisation of the elevated “zero-maze” as an animal model of anxiety. Psychopharmacology (Berl) 116: 56–64.
Suzuki, T., M. Inayama, and M. Misawa. 1990. The effect of diazepam on exploratory behavior and its strain differences in inbred rats. Yakubutsu, Seishin, Kodo 10: 307–314.
Kaesermann, H.P. 1986. Stretched attend posture, a non-social form of ambivalence, is sensitive to a conflict-reducing drug action. Psychopharmacology (Berl) 89: 31–37.
Dantzer, R., R.M. Bluthe, G.F. Koob, and M. Le Moal. 1987. Modulation of social memory in male rats by neurohypophyseal peptides. Psychopharmacology (Berl) 91: 363–368.
Letty, S., R. Child, A. Dumuis, A. Pantaloni, J. Bockaert, and G. Rondouin. 1997. 5-HT4 receptors improve social olfactory memory in the rat. Neuropharmacology 36: 681–687.
Sawyer, T.F., A.K. Hengehold, and W.A. Perez. 1984. Chemosensory and hormonal mediation of social memory in male rats. Behavioral Neuroscience 98: 908–913.
Dimitrova, D.S., and D.P. Getova-Spassova. 2006. Effects of galantamine and donepezil on active and passive avoidance tests in rats with induced hypoxia. Journal of Pharmacological Sciences 101: 199–204.
Faridkia, Z., Yaghmaei, P., Nassiri-Asl, M. 2016. Protective effect of quinine on chemical kindling and passive avoidance test in rats. Iran Red Crescent Med J 18: e25490.
Paxinos, G., and C. Watson. 2002. The rat brain in stereotaxic coordinates, 4th ed. New York: Academic Press.
Livak, K.J., and T.D. Schmittgen. 2001. Analysis of relative gene expression data using real-time quantitative PCR and the 2(-Delta Delta C(T)) method. Methods 25: 402–408.
Brand, D.D., K.A. Latham, and E.F. Rosloniec. 2007. Collagen-induced arthritis. Nature Protocols 2: 1269–1275.
Bendele, A. 2001. Animal models of rheumatoid arthritis. Journal of Musculoskeletal and Neuronal Interactions 1: 377–385.
Sengupta, P. 2013. The laboratory rat: Relating its age with human’s. IInt J Prev Med 4: 624–630.
Verazza, S., G. Negro, D. Marafon, A. Consolaro, A. Martini, and A. Ravelli. 2013. Possible discontinuation of therapies after clinical remission in juvenile idiopathic arthritis. Clinical and Experimental Rheumatology 31: S98-101.
Jiang, P., Q. Ling, H. Liu, and W. Tu. 2015. Intracisternal administration of an interleukin-6 receptor antagonist attenuates surgery-induced cognitive impairment by inhibition of neuroinflammatory responses in aged rats. Experimental and Therapeutic Medicine 9: 982–986.
Elcioglu, H.K., E. Aslan, S. Ahmad, S. Alan, E. Salva, O.H. Elcioglu, and L. Kabasakal. 2016. Tocilizumab’s effect on cognitive deficits induced by intracerebroventricular administration of streptozotocin in Alzheimer’s model. Molecular and Cellular Biochemistry 420: 21–28.
Stone, R. H., Hong, J., Jeong, H. 2014. Pharmacokinetics of monoclonal antibodies used for inflammatory bowel diseases in pregnant women. J Clin Toxicol. 4.
Luheshi, G.N., R.M. Bluthe, D. Rushforth, N. Mulcahy, J.P. Konsman, M. Goldbach, and R. Dantzer. 2000. Vagotomy attenuates the behavioural but not the pyrogenic effects of interleukin-1 in rats. Autonomic Neuroscience 85: 127–132.
Wiener, C.D., F.P. Moreira, L.V. Portela, N.R. Strogulski, D.R. Lara, R. da Silva, L.D. de Souza, and M., Jansen, K., Oses, J. P. . 2019. Interleukin-6 and Interleukin-10 in mood disorders: A population-based study. Psychiatry Research 273: 685–689.
Muller, N., and M.J. Schwarz. 2007. The immune-mediated alteration of serotonin and glutamate: Towards an integrated view of depression. Molecular Psychiatry 12: 988–1000.
Nishioku, T., A. Yamauchi, F. Takata, T. Watanabe, K. Furusho, H. Shuto, A. Yamauchi, and Y. Kataoka. 2010. Disruption of the blood-brain barrier in collagen-induced arthritic mice. Neuroscience Letters 482: 208–211.
Nishioku, T., K. Furusho, A. Tomita, H. Ohishi, S. Dohgu, H. Shuto, A. Yamautsi, and Y. Kataoka. 2011. Potential role for S100A4 in the disruption of the blood-brain barrier in collagen-induced arthritic mice, an animal model of rheumatoid arthritis. Neuroscience 189: 286–292.
Mokotedi, L., Millen, A. M. E., Mogane, C., Gomes, M., Woodiwiss, A. J., Norton, G. R., Michel, F. S. 2019. Associations of inflammatory markers and vascular cell adhesion molecule-1 with endothelial dysfunction in collagen-induced arthritis. Eur J Pharmacol 865: 172786.
Lassman, A.B., L.E. Abrey, G.D. Shah, K.S. Panageas, M. Begemann, M.G. Malkin, and J.J. Raizer. 2006. Systemic high-dose intravenous methotrexate for central nervous system metastases. Journal of Neuro-oncology 78: 255–260.
Murinova, J., N. Hlavacova, M. Chmelova, and I. Riecansky. 2017. The evidence for altered BDNF expression in the brain of rats reared or housed in social isolation: A systematic review. Frontiers in Behavioral Neuroscience 11: 101.
Yang, M., J. Kim, S.H. Kim, J.S. Kim, T. Shin, and C. Moon. 2012. Temporal profiles of synaptic plasticity-related signals in adult mouse hippocampus with methotrexate treatment. Neural Regeneration Research 7: 1651–1658.
ACKNOWLEDGEMENTS
We thank Athanasia Pappa, president of the Hellenic League Against Rheumatism, for her contribution during various stages of the research project. We also thank Roche Hellas for providing tocilizumab.
Author information
Authors and Affiliations
Contributions
F.P., A.S. and F.M. performed the in vivo and in vitro experiments. F.P. drafted the original manuscript and conducted the statistical analysis. She is the guarantor. C.P. contributed to the conception and design of the study, reviewed and edited the manuscript. M.E.M. performed the histological examination of the samples. D.K. contributed to the conception and design of the study and supervised the project. All authors critically reviewed and approved the final manuscript. All authors agree to be accountable for the work and to ensure that any questions relating to the accuracy and integrity of the paper are investigated and properly resolved.
Corresponding author
Ethics declarations
Ethics Approval
The study was approved by the Directorate of Veterinary Services of the Region of Central Macedonia according to national legislation (Presidential Decree 56/2013, in conformance with the European Directive (2010/63/EU) (reference number: 668476(3484) 21643(87), 06/10/2019).
Consent to Participate
Not applicable.
Consent for Publication
Not applicable.
Conflict of Interest
The authors declare no competing interests.
Additional information
Publisher's Note
Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.
Rights and permissions
About this article
Cite this article
Poutoglidou, F., Pourzitaki, C., Manthou, M.E. et al. Infliximab and Tocilizumab Reduce Anxiety-Like Behaviour and Improve Cognitive Performance in a Juvenile Collagen–Induced Arthritis Rat Model. Inflammation 45, 445–459 (2022). https://doi.org/10.1007/s10753-021-01560-6
Received:
Accepted:
Published:
Issue Date:
DOI: https://doi.org/10.1007/s10753-021-01560-6