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Journal of Neuroinflammation

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Open Access 01-12-2019 | Stroke | Research

Augmented β2-adrenergic signaling dampens the neuroinflammatory response following ischemic stroke and increases stroke size

Authors: Kendra J. Lechtenberg, Scott T. Meyer, Janelle B. Doyle, Todd C. Peterson, Marion S. Buckwalter

Published in: Journal of Neuroinflammation | Issue 1/2019

Abstract

Background

Ischemic stroke provokes a neuroinflammatory response and simultaneously promotes release of epinephrine and norepinephrine by the sympathetic nervous system. This increased sympathetic outflow can act on β2-adrenergic receptors expressed by immune cells such as brain-resident microglia and monocyte-derived macrophages (MDMs), but the effect on post-stroke neuroinflammation is unknown. Thus, we investigated how changes in β2-adrenergic signaling after stroke onset influence the microglia/MDM stroke response, and the specific importance of microglia/MDM β2-adrenergic receptors to post-stroke neuroinflammation.

Methods

To investigate the effects of β2-adrenergic receptor manipulation on post-stroke neuroinflammation, we administered the β2-adrenergic receptor agonist clenbuterol to mice 3 h after the onset of photothrombotic stroke. We immunostained to quantify microglia/MDM numbers and proliferation and to assess morphology and activation 3 days later. We assessed stroke outcomes by measuring infarct volume and functional motor recovery and analyzed gene expression levels of neuroinflammatory molecules. Finally, we evaluated changes in cytokine expression and microglia/MDM response in brains of mice with selective knockout of the β2-adrenergic receptor from microglia and monocyte-lineage cells.

Results

We report that clenbuterol treatment after stroke onset causes enlarged microglia/MDMs and impairs their proliferation, resulting in reduced numbers of these cells in the peri-infarct cortex by 1.7-fold at 3 days after stroke. These changes in microglia/MDMs were associated with increased infarct volume in clenbuterol-treated animals. In mice that had the β2-adrenergic receptor specifically knocked out of microglia/MDMs, there was no change in morphology or numbers of these cells after stroke. However, knockdown of β2-adrenergic receptors in microglia and MDMs resulted in increased expression of TNFα and IL-10 in peri-infarct tissue, while stimulation of β2-adrenergic receptors with clenbuterol had the opposite effect, suppressing TNFα and IL-10 expression.

Conclusions

We identified β2-adrenergic receptor signaling as an important regulator of the neuroimmune response after ischemic stroke. Increased β2-adrenergic signaling after stroke onset generally suppressed the microglia/MDM response, reducing upregulation of both pro- and anti-inflammatory cytokines, and increasing stroke size. In contrast, diminished β2-adrenergic signaling in microglia/MDMs augmented both pro- and anti-inflammatory cytokine expression after stroke. The β2-adrenergic receptor may therefore present a therapeutic target for improving the post-stroke neuroinflammatory and repair process.

Iadecola C, Anrather J. The immunology of stroke: from mechanisms to translation. Nat Med. 2011;17:796–808. https://doi.org/10.1038/nm.2399.CrossRefPubMedPubMedCentral

Kim E, Cho S. Microglia and monocyte-derived macrophages in stroke. Neurotherapeutics. 2016;13:702–18. https://doi.org/10.1007/s13311-016-0463-1.CrossRefPubMedPubMedCentral

Meyer JS, Stoica E, Pascu I, Shimazu K, Hartmann A. Catecholamine concentrations in CSF and plasma of patients with cerebral infarction and haemorrhage. Brain. 1973;96:277–88.CrossRefPubMed

Sander D, Winbeck K, Klingelhöfer J, Etgen T, Conrad B. Prognostic relevance of pathological sympathetic activation after acute thromboembolic stroke. Neurology. 2001;57:833–8.CrossRefPubMed

Mracsko E, Liesz A, Karcher S, Zorn M, Bari F, Veltkamp R. Differential effects of sympathetic nervous system and hypothalamic-pituitary-adrenal axis on systemic immune cells after severe experimental stroke. Brain Behav Immun. 2014;41:200–9.CrossRefPubMed

Jauch EC, Saver JL, Adams HP, Bruno A, Connors JJB, Demaerschalk BM, et al. Guidelines for the early management of patients with acute ischemic stroke: a guideline for healthcare professionals from the American Heart Association/American Stroke Association. Stroke. 2013;44:870–947.CrossRefPubMed

Marino F, Cosentino M. Adrenergic modulation of immune cells: an update. Amino Acids. 2013;45:55–71.CrossRefPubMed

Prass K, Meisel C, Höflich C, Braun J, Halle E, Wolf T, et al. Stroke-induced immunodeficiency promotes spontaneous bacterial infections and is mediated by sympathetic activation reversal by poststroke T helper cell type 1-like immunostimulation. J Exp Med. 2003;198:725–36. https://doi.org/10.1084/jem.20021098.CrossRefPubMedPubMedCentral

McCulloch L, Smith CJ, McColl BW. Adrenergic-mediated loss of splenic marginal zone B cells contributes to infection susceptibility after stroke. Nat Commun. 2017;8:15051. https://doi.org/10.1038/ncomms15051.CrossRefPubMedPubMedCentral

10.

Semkova I, Schilling M, Henrich-Noack P, Rami A, Krieglstein J. Clenbuterol protects mouse cerebral cortex and rat hippocampus from ischemic damage and attenuates glutamate neurotoxicity in cultured hippocampal neurons by induction of NGF. Brain Res. 1996;717:44–54 http://www.ncbi.nlm.nih.gov/pubmed/8738252.CrossRefPubMed

11.

Culmsee C, Semkova I, Krieglstein J. NGF mediates the neuroprotective effect of the beta2-adrenoceptor agonist clenbuterol in vitro and in vivo: evidence from an NGF-antisense study. Neurochem Int. 1999;35:47–57 http://www.ncbi.nlm.nih.gov/pubmed/10403429.CrossRefPubMed

12.

Culmsee C, Stumm RK, Schäfer MK, Weihe E, Krieglstein J. Clenbuterol induces growth factor mRNA, activates astrocytes, and protects rat brain tissue against ischemic damage. Eur J Pharmacol. 1999;379:33–45 http://www.ncbi.nlm.nih.gov/pubmed/10499369.CrossRefPubMed

13.

Han RQ, Ouyang YB, Xu L, Agrawal R, Patterson AJ, Giffard RG. Postischemic brain injury is attenuated in mice lacking the β2-adrenergic receptor. Anesth Analg. 2009;108:280–7.CrossRefPubMedPubMedCentral

14.

White RE, Palm C, Xu L, Ling E, Ginsburg M, Daigle BJ, et al. Mice lacking the β2 adrenergic receptor have a unique genetic profile before and after focal brain ischaemia. ASN Neuro. 2012;4:343–56.CrossRef

15.

Wattananit S, Tornero D, Graubardt N, Memanishvili T, Monni E, Tatarishvili J, et al. Monocyte-derived macrophages contribute to spontaneous long-term functional recovery after stroke in mice. J Neurosci. 2016;36:4182–95. https://doi.org/10.1523/JNEUROSCI.4317-15.2016.CrossRefPubMedPubMedCentral

16.

Gelderblom M, Leypoldt F, Steinbach K, Behrens D, Choe CU, Siler DA, et al. Temporal and spatial dynamics of cerebral immune cell accumulation in stroke. Stroke. 2009;40:1849–57.CrossRefPubMed

17.

Radojcic T, Baird S, Darko D, Smith D, Bulloch K. Changes in β-adrenergic receptor distribution on immunocytes during differentiation: an analysis of T cells and macrophages. J Neurosci Res. 1991;30:328–35.CrossRefPubMed

18.

Mori K, Ozaki E, Zhang B, Yang L, Yokoyama A, Takeda I, et al. Effects of norepinephrine on rat cultured microglial cells that express alpha1, alpha2, beta1 and beta2 adrenergic receptors. Neuropharmacology. 2002;43:1026–34.CrossRefPubMed

19.

Prinz M, Häusler KG, Kettenmann H. Hanisch UK. β-adrenergic receptor stimulation selectively inhibits IL-12p40 release in microglia. Brain Res. 2001;899:264–70.CrossRefPubMed

20.

Zhang Y, Chen K, Sloan SA, Bennett ML, Scholze AR, Keeffe SO, et al. An RNA-sequencing transcriptome and splicing database of glia, neurons, and vascular cells of the cerebral cortex. J Neurosci. 2014;34:11929–47.CrossRefPubMedPubMedCentral

21.

Ishii Y, Yamaizumi A, Kawakami A, Islam A, Choudhury ME, Takahashi H, et al. Anti-inflammatory effects of noradrenaline on LPS-treated microglial cells: suppression of NFκB nuclear translocation and subsequent STAT1 phosphorylation. Neurochem Int. 2015;90:56–66. https://doi.org/10.1016/j.neuint.2015.07.010.CrossRefPubMed

22.

Dello Russo C, Boullerne AI, Gavrilyuk V, Feinstein DL. Inhibition of microglial inflammatory responses by norepinephrine: effects on nitric oxide and interleukin-1β production. J Neuroinflammation. 2004;1:9.CrossRef

23.

Sharma M, Flood PM. β-arrestin2 regulates the anti-inflammatory effects of salmeterol in lipopolysaccharide-stimulated BV2 cells. J Neuroimmunol. 2018;325(July):10–9. https://doi.org/10.1016/j.jneuroim.2018.10.001.CrossRefPubMed

24.

Färber K, Pannasch U, Kettenmann H. Dopamine and noradrenaline control distinct functions in rodent microglial cells. Mol Cell Neurosci. 2005;29:128–38.CrossRefPubMed

25.

Jiang L, Chen SH, Chu CH, Wang SJ, Oyarzabal E, Wilson B, et al. A novel role of microglial NADPH oxidase in mediating extra-synaptic function of norepinephrine in regulating brain immune homeostasis. Glia. 2015;63:1057–72. https://doi.org/10.1002/glia.22801.CrossRefPubMedPubMedCentral

26.

Heneka MT, Nadrigny F, Regen T, Martinez-Hernandez A, Dumitrescu-Ozimek L, Terwel D, et al. Locus ceruleus controls Alzheimer’s disease pathology by modulating microglial functions through norepinephrine. Proc Natl Acad Sci U S A. 2010;107:6058–63. https://doi.org/10.1073/pnas.0909586107.CrossRefPubMedPubMedCentral

27.

Parkhurst CN, Yang G, Ninan I, Savas JN, Yates JR, Lafaille JJ, et al. Microglia promote learning-dependent synapse formation through BDNF. Cell. 2013;155:1596–609. https://doi.org/10.1016/j.cell.2013.11.030.CrossRefPubMedPubMedCentral

28.

Labat-gest V, Tomasi S. Photothrombotic ischemia: a minimally invasive and reproducible photochemical cortical lesion model for mouse stroke studies. J Vis Exp. 2013:1–6. https://doi.org/10.3791/50370.

29.

Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 22DDCT method. Methods. 2001;25:402–8.CrossRefPubMed

30.

Schaar KL, Brenneman MM, Savitz SI. Functional assessments in the rodent stroke model. Exp Transl Stroke Med. 2010;2:1–11.CrossRef

31.

Cheng MY, Aswendt M, Steinberg GK. Optogenetic approaches to target specific neural circuits in post-stroke recovery. Neurotherapeutics. 2016;13:325–40. https://doi.org/10.1007/s13311-015-0411-5.CrossRefPubMed

32.

Brown R, Motulsky H. Detecting outliers when fitting data with nonlinear regression - a new method based on robust nonlinear regression and the false discovery rate. BMC Bioinformatics. 2006;7:1–20. https://doi.org/10.1186/1471-2105-7-123.CrossRef

33.

Urra X, Villamor N, Planas AM. Harms and benefits of lymphocyte subpopulations in patients with acute stroke. NSC. 2009;158:1174–83. https://doi.org/10.1016/j.neuroscience.2008.06.014.CrossRef

34.

Armstrong L, Jordan N, Millar A. Interleukin 10 (IL-10) regulation of tumour necrosis factor α ( TNF-α) from human alveolar macrophages and peripheral blood monocytes. Thorax. 1996;51:143–9.CrossRefPubMedPubMedCentral

35.

Eizenberg Y, Grossman E, Tanne D, Koton S. Pre admission treatment with beta-blockers in hypertensive patients with acute stroke and 3-month outcome—data from a national stroke registry. J Clin Hypertens. 2018;20:568–72.CrossRef

36.

Maier I, Becker J, Leyhe J, Schnieder M, Behme D, Psychogios M-K, et al. Influence of beta-blocker therapy on the risk of infections and death in patients at high risk for stroke induced immunodepression. PLoS One. 2018;13. https://doi.org/10.1371/journal.pone.0196174.

37.

Sykora M, Siarnik P, Diedler J, Lees KR, Alexandrov A, Bath PM, et al. β-Blockers, pneumonia, and outcome after ischemic stroke: evidence from virtual international stroke trials archive. Stroke. 2015;46:1269–74. https://doi.org/10.1161/STROKEAHA.114.008260.CrossRefPubMed

38.

Westendorp WF, Nederkoorn PJ, Vermeij J-D, Dijkgraaf MG, van de Beek D. Post-stroke infection: a systematic review and meta-analysis. BMC Neurol. 2011;11:110. https://doi.org/10.1186/1471-2377-11-110.CrossRefPubMedPubMedCentral

39.

Shim R, Wong CHY. Complex interplay of multiple biological systems that contribute to post-stroke infections. Brain Behav Immun. 2018;70:10–20. https://doi.org/10.1016/j.bbi.2018.03.019.CrossRefPubMed

40.

Zhu Y, Culmsee C, Semkova I, Krieglstein J. Stimulation of beta2-adrenoceptors inhibits apoptosis in rat brain after transient forebrain ischemia. J Cereb Blood Flow Metab. 1998;18:1032–9. https://doi.org/10.1097/00004647-199809000-00013.CrossRefPubMed

41.

Wang H, Deng QW, Peng AN, Xing FL, Zuo L, Li S, et al. β-arrestin2 functions as a key regulator in the sympathetic-triggered immunodepression after stroke. J Neuroinflammation. 2018;15:1–11.CrossRef

42.

Gyoneva S, Traynelis SF. Norepinephrine modulates the motility of resting and activated microglia via different adrenergic receptors. J Biol Chem. 2013;288:15291–302.CrossRefPubMedPubMedCentral

43.

Fujita H, Tanaka J, Maeda N, Sakanaka M. Adrenergic agonists suppress the proliferation of microglia through β2- adrenergic receptor. Neurosci Lett. 1998;242:37–40.CrossRefPubMed

44.

Moriyama S, Brestoff JR, Flamar A-L, Moeller JB, Klose CSN, Rankin LC, et al. β2-adrenergic receptor-mediated negative regulation of group 2 innate lymphoid cell responses. Science. 2018;359:1056–61.CrossRefPubMed

45.

Ryan KJ, Griffin É, Yssel JD, Ryan KM, McNamee EN, Harkin A, et al. Stimulation of central β2-adrenoceptors suppresses NFκB activity in rat brain: a role for IκB. Neurochem Int. 2013;63:368–78. https://doi.org/10.1016/j.neuint.2013.07.006.CrossRefPubMed

46.

Gleeson LC, Ryan KJ, Griffin EW, Connor TJ, Harkin A. The β2-adrenoceptor agonist clenbuterol elicits neuroprotective, anti-inflammatory and neurotrophic actions in the kainic acid model of excitotoxicity. Brain Behav Immun. 2010;24:1354–61. https://doi.org/10.1016/j.bbi.2010.06.015.CrossRefPubMed

47.

Szalay G, Martinecz B, Lénárt N, Környei Z, Orsolits B, Judák L, et al. Microglia protect against brain injury and their selective elimination dysregulates neuronal network activity after stroke. Nat Commun. 2016;7. https://doi.org/10.1038/ncomms11499.

48.

Shichita T, Ito M, Morita R, Komai K, Noguchi Y, Ooboshi H, et al. MAFB prevents excess inflammation after ischemic stroke by accelerating clearance of damage signals through MSR1. Nat Med. 2017;23:723–32. https://doi.org/10.1038/nm.4312.CrossRefPubMed

49.

McNamee EN, Ryan KM, Griffin EW, González-Reyes RE, Ryan KJ, Harkin A, et al. Noradrenaline acting at central beta-adrenoceptors induces interleukin-10 and suppressor of cytokine signaling-3 expression in rat brain: implications for neurodegeneration. Brain Behav Immun. 2010;24:660–71. https://doi.org/10.1016/j.bbi.2010.02.005.CrossRefPubMed

50.

Giordani L, Cuzziol N, Del Pinto T, Sanchez M, Maccari S, Massimi A, et al. β2-agonist clenbuterol hinders human monocyte differentiation into dendritic cells. Exp Cell Res. 2015;339:163–73. https://doi.org/10.1016/j.yexcr.2015.10.032.CrossRefPubMed

51.

Ağaç D, Estrada LD, Maples R, Hooper LV, Farrar JD. The β2-adrenergic receptor controls inflammation by driving rapid IL-10 secretion. Brain Behav Immun. 2018;74:176–85.CrossRefPubMedPubMedCentral

52.

Platzer C, Meisel C, Vogt K, Platzer M, Volk HD. Up-regulation of monocytic IL-10 by tumor necrosis factor-α and cAMP elevating drugs. Int Immunol. 1995;7:517–23.CrossRefPubMed

53.

Sato TA, Keelan JA, Mitchell MD. Critical paracrine interactions between TNF-alpha and IL-10 regulate lipopolysaccharide-stimulated human choriodecidual cytokine and prostaglandin E2 production. J Immunol. 2003;170:158–66. https://doi.org/10.4049/jimmunol.170.1.158.CrossRefPubMed

54.

Meng A, Wang B, Zhang X, Qi N, Liu D, Wu J. Additive suppression of LPS-induced IL-10 and TNF-α by pre-treatment of dexamethasone and SB203580 in a murine alveolar macrophage cell line (MH-S). Inflammation. 2015;38:1260–6.CrossRefPubMed

55.

Farmer P, Pugin J. β-Adrenergic agonists exert their “anti-inflammatory” effects in monocytic cells through the IκB/NF-κB pathway. Am J Physiol Lung Cell Mol Physiol. 2000;279:675–82.CrossRef

56.

Nance DM, Sanders VM. Autonomic innervation and regulation of the immune system (1987-2007). Brain Behav Immun. 2007;21:736–45. https://doi.org/10.1016/j.bbi.2007.03.008.CrossRefPubMedPubMedCentral

57.

Izeboud CA, Monshouwer M, Van MASJPAM, Witkamp RF. The β-adrenoceptor agonist clenbuterol is a potent inhibitor of the LPS-induced production of TNF-α and IL-6 in vitro and in vivo. Inflamm Res. 1999;48:497–502.CrossRefPubMed

58.

Lorton D, Bellinger DL. Molecular mechanisms underlying β-adrenergic receptor-mediated cross-talk between sympathetic neurons and immune cells. Int J Mol Sci. 2015;16:5635–65.CrossRefPubMedPubMedCentral

59.

Kolmus K, Tavernier J, Gerlo S. β2-adrenergic receptors in immunity and inflammation: stressing NF-κB. Brain Behav Immun. 2015;45:297–310. https://doi.org/10.1016/j.bbi.2014.10.007.CrossRefPubMed

60.

Szelenyi J, Selmeczy Z, Brozik A, Medgyesi D, Magocsi M. Dual β-adrenergic modulation in the immune system: stimulus-dependent effect of isoproterenol on MAPK activation and inflammatory mediator production in macrophages. Neurochem Int. 2006;49:94–103.CrossRefPubMed

61.

Lefkowitz RJ. G protein-coupled receptors. J Biol Chem. 1998;273:18677–80.CrossRefPubMed

62.

Stanojević S, Dimitrijević M, Kuštrimović N, Mitić K, Vujić V, Leposavić G. Adrenal hormone deprivation affects macrophage catecholamine metabolism and β2-adrenoceptor density, but not propranolol stimulation of tumour necrosis factor-α production. Exp Physiol. 2013;98:665–78.CrossRefPubMed

Title: Augmented β2-adrenergic signaling dampens the neuroinflammatory response following ischemic stroke and increases stroke size
Authors: Kendra J. Lechtenberg
Scott T. Meyer
Janelle B. Doyle
Todd C. Peterson
Marion S. Buckwalter
Publication date: 01-12-2019
Publisher: BioMed Central
Keywords: Stroke
Clenbuterol
Published in: Journal of Neuroinflammation / Issue 1/2019
Electronic ISSN: 1742-2094
DOI: https://doi.org/10.1186/s12974-019-1506-4

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Augmented β2-adrenergic signaling dampens the neuroinflammatory response following ischemic stroke and increases stroke size

Abstract

Background

Methods

Results

Conclusions

Keynote webinar | Spotlight on medication adherence

Springer Medicine

Abstract

Background

Methods

Results

Conclusions

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