Inflammation Research

, Volume 65, Issue 11, pp 869–879 | Cite as

The cannabinoid 2 receptor agonist β-caryophyllene modulates the inflammatory reaction induced by Mycobacterium bovis BCG by inhibiting neutrophil migration

  • Magaiver Andrade-Silva
  • Luana Barbosa Correa
  • André Luis Peixoto Candéa
  • Simone C. Cavalher-Machado
  • Helene Santos Barbosa
  • Elaine Cruz Rosas
  • Maria G HenriquesEmail author
Original Research Paper


Objective and design

β-Caryophyllene (BCP) is a sesquiterpene that binds to the cannabinoid 2 (CB2) receptor and exerts anti-inflammatory effects. In this study, we investigated the anti-inflammatory effect of BCP and another CB2 agonist, GP1a in inflammatory experimental model induced by Mycobacterium bovis (BCG).


C57Bl/6 mice were pretreated orally with BCP (0.5–50 mg/kg) or intraperitonealy with GP1a (10 mg/kg) 1 h before the induction of pleurisy or pulmonary inflammation by BCG. The direct action of CB2 agonists on neutrophils function was evaluated in vitro.


β-Caryophyllene (50 mg/kg) impaired BCG-induced neutrophil accumulation in pleurisy without affecting mononuclear cells or the production of TNF-α and CCL2/MCP-1. However, BCP inhibited CXCL1/KC, leukotriene B4 (LTB4), IL-12, and nitric oxide production. GP1a had a similar effect to BCP. Preincubation of neutrophils with BCP (10 µM) impaired chemotaxis toward LTB4 and adhesion to endothelial cells stimulated with TNF-α, and both, BCP and GP1a, impaired LTB4-induced actin polymerization.


These results suggest that the CB2 receptor may represent a new target for modulating the inflammatory reaction induced by mycobacteria.


CB2 receptor GP1a Neutrophil Mycobacteria Chemotaxis 



This work was supported by the Fundação Carlos Chagas Filho de Amparo a Pesquisa do Estado do Rio de Janeiro (FAPERJ) Sediadas 32/2013), Conselho Nacional de Desenvolvimento Científico e Tecnologico (CNPq) 305986/2014-7 and Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES) PROCAD 149/2007. M. A. Silva and L. B. Correa are students of the Programa de Pós-Graduação em Biologia Celular e Molecular from Instituto Oswaldo Cruz, FIOCRUZ, Rio de Janeiro, Brazil.

Compliance with ethical standards

Conflict of interest

The authors declare no conflicts of interests.

Supplementary material

11_2016_969_MOESM1_ESM.pdf (567 kb)
Supplementary material 1 (PDF 566 kb)
11_2016_969_MOESM2_ESM.pdf (367 kb)
Supplementary material 2 (PDF 367 kb)


  1. 1.
    Gertsch J, Leonti M, Raduner S, Racz I, Chen JZ, Xie XQ, et al. Beta-caryophyllene is a dietary cannabinoid. Proc Natl Acad Sci USA. 2008;105:9099–104.CrossRefPubMedPubMedCentralGoogle Scholar
  2. 2.
    Astani A, Reichling J, Schnitzler P. Screening for antiviral activities of isolated compounds from essential oils. Evid Based Complement Altern Med. 2011;2011:253643.CrossRefGoogle Scholar
  3. 3.
    Basha RH, Sankaranarayanan C. β-Caryophyllene, a natural sesquiterpene lactone attenuates hyperglycemia mediated oxidative and inflammatory stress in experimental diabetic rats. Chem Biol Interact. 2016;245:50–8.CrossRefPubMedGoogle Scholar
  4. 4.
    Hammami S, Jmii H, Mokni RE, Khmiri A, Faidi K, Dhaouadi H, et al. Essential oil composition, antioxidant, cytotoxic and antiviral activities of teucrium pseudochamaepitys growing spontaneously in Tunisia. Molecules. 2015;20:20426–33.CrossRefPubMedGoogle Scholar
  5. 5.
    Asdadi A, Hamdouch A, Oukacha A, Moutaj R, Gharby S, Harhar H, et al. Study on chemical analysis, antioxidant and in vitro antifungal activities of essential oil from wild Vitex agnus-castus L. seeds growing in area of argan tree of Morocco against clinical strains of Candida responsible for nosocomial infections. J Mycol Med. 2015;25:e118–27.CrossRefPubMedGoogle Scholar
  6. 6.
    Klauke AL, Racz I, Pradier B, Markert A, Zimmer AM, Gertsch J, et al. The cannabinoid CB2 receptor-selective phytocannabinoid beta-caryophyllene exerts analgesic effects in mouse models of inflammatory and neuropathic pain. Eur Neuropsychopharmacol. 2014;24:608–20.CrossRefPubMedGoogle Scholar
  7. 7.
    Fernandes ES, Passos GF, Medeiros R, da Cunha FM, Ferreira J, Campos MM, et al. Anti-inflammatory effects of compounds alpha-humulene and (−)-trans-caryophyllene isolated from the essential oil of Cordia verbenacea. Eur J Pharmacol. 2007;569:228–36.CrossRefPubMedGoogle Scholar
  8. 8.
    Singh UP, Singh NP, Singh B, Price RL, Nagarkatti M, Nagarkatti PS. Cannabinoid receptor-2 (CB2) agonist ameliorates colitis in IL-10(−/−) mice by attenuating the activation of T cells and promoting their apoptosis. Toxicol Appl Pharmacol. 2012;258:256–67.CrossRefPubMedGoogle Scholar
  9. 9.
    Tschöp J, Kasten KR, Nogueiras R, Goetzman HS, Cave CM, England LG, et al. The cannabinoid receptor 2 is critical for the host response to sepsis. J Immunol. 2009;183:499–505.CrossRefPubMedPubMedCentralGoogle Scholar
  10. 10.
    Murikinati S, Jüttler E, Keinert T, Ridder DA, Muhammad S, Waibler Z, et al. Activation of cannabinoid 2 receptors protects against cerebral ischemia by inhibiting neutrophil recruitment. FASEB J. 2010;24:788–98.CrossRefPubMedGoogle Scholar
  11. 11.
    Klein TW. Cannabinoid-based drugs as anti-inflammatory therapeutics. Nat Rev Immunol. 2005;5:400–11.CrossRefPubMedGoogle Scholar
  12. 12.
    Toguri JT, Lehmann C, Laprairie RB, Szczesniak AM, Zhou J, Denovan-Wright EM, et al. Anti-inflammatory effects of cannabinoid CB(2) receptor activation in endotoxin-induced uveitis. Br J Pharmacol. 2014;171:1448–61.CrossRefPubMedPubMedCentralGoogle Scholar
  13. 13.
    Fukuda S, Kohsaka H, Takayasu A, Yokoyama W, Miyabe C, Miyabe Y, et al. Cannabinoid receptor 2 as a potential therapeutic target in rheumatoid arthritis. BMC Musculoskelet Disord. 2014;15:275.CrossRefPubMedPubMedCentralGoogle Scholar
  14. 14.
    Smith SR, Denhardt G, Terminelli C. The anti-inflammatory activities of cannabinoid receptor ligands in mouse peritonitis models. Eur J Pharmacol. 2001;432:107–19.CrossRefPubMedGoogle Scholar
  15. 15.
    Bento AF, Marcon R, Dutra RC, Claudino RF, Cola M, Pereira Leite DF, et al. β-Caryophyllene inhibits dextran sulfate sodium-induced colitis in mice through CB2 receptor activation and PPARγ pathway. Am J Pathol. 2011;178:1153–66.CrossRefPubMedPubMedCentralGoogle Scholar
  16. 16.
    Horváth B, Mukhopadhyay P, Kechrid M, Patel V, Tanchian G, Wink DA, et al. β-Caryophyllene ameliorates cisplatin-induced nephrotoxicity in a cannabinoid 2 receptor-dependent manner. Free Radic Biol Med. 2012;52:1325–33.CrossRefPubMedPubMedCentralGoogle Scholar
  17. 17.
    Lawn SD, Zumla AI. Tuberculosis. Lancet. 2011;378:57–72.CrossRefPubMedGoogle Scholar
  18. 18.
    O’Garra A, Redford PS, McNab FW, Bloom CI, Wilkinson RJ, Berry MP. The immune response in tuberculosis. Annu Rev Immunol. 2013;31:475–527.CrossRefPubMedGoogle Scholar
  19. 19.
    Berrington WR, Hawn TR. Mycobacterium tuberculosis, macrophages, and the innate immune response: does common variation matter? Immunol Rev. 2007;219:167–86.CrossRefPubMedPubMedCentralGoogle Scholar
  20. 20.
    Antony VB, Sahn SA, Antony AC, Repine JE. Bacillus Calmette–Guérin-stimulated neutrophils release chemotaxins for monocytes in rabbit pleural spaces and in vitro. J Clin Invest. 1985;76:1514–21.CrossRefPubMedPubMedCentralGoogle Scholar
  21. 21.
    Kasahara K, Sato I, Ogura K, Takeuchi H, Kobayashi K, Adachi M. Expression of chemokines and induction of rapid cell death in human blood neutrophils by Mycobacterium tuberculosis. J Infect Dis. 1998;178:127–37.CrossRefPubMedGoogle Scholar
  22. 22.
    Menezes-de-Lima-Júnior O, Werneck-Barroso E, Cordeiro RS, Henriques MG. Effects of inhibitors of inflammatory mediators and cytokines on eosinophil and neutrophil accumulation induced by Mycobacterium bovis bacillus Calmette–Guérin in mouse pleurisy. J Leukoc Biol. 1997;62:778–85.PubMedGoogle Scholar
  23. 23.
    Souza MC, Penido C, Costa MF, Henriques MG. Mechanisms of T-lymphocyte accumulation during experimental pleural infection induced by Mycobacterium bovis BCG. Infect Immun. 2008;76:5686–93.CrossRefPubMedPubMedCentralGoogle Scholar
  24. 24.
    Penido C, Vieira-de-Abreu A, Bozza MT, Castro-Faria-Neto HC, Bozza PT. Role of monocyte chemotactic protein-1/CC chemokine ligand 2 on gamma delta T lymphocyte trafficking during inflammation induced by lipopolysaccharide or Mycobacterium bovis bacille Calmette–Guérin. J Immunol. 2003;171:6788–94.CrossRefPubMedGoogle Scholar
  25. 25.
    Costa MF, de Souza-Martins R, de Souza MC, Benjamim CF, Piva B, Diaz BL, et al. Leukotriene B4 mediates gammadelta T lymphocyte migration in response to diverse stimuli. J Leukoc Biol. 2010;87:323–32.CrossRefPubMedGoogle Scholar
  26. 26.
    Eruslanov EB, Lyadova IV, Kondratieva TK, Majorov KB, Scheglov IV, Orlova MO, et al. Neutrophil responses to Mycobacterium tuberculosis infection in genetically susceptible and resistant mice. Infect Immun. 2005;73:1744–53.CrossRefPubMedPubMedCentralGoogle Scholar
  27. 27.
    D’Avila H, Roque NR, Cardoso RM, Castro-Faria-Neto HC, Melo RC, Bozza PT. Neutrophils recruited to the site of Mycobacterium bovis BCG infection undergo apoptosis and modulate lipid body biogenesis and prostaglandin E production by macrophages. Cell Microbiol. 2008;10:2589–604.CrossRefPubMedGoogle Scholar
  28. 28.
    Marzo E, Vilaplana C, Tapia G, Diaz J, Garcia V, Cardona PJ. Damaging role of neutrophilic infiltration in a mouse model of progressive tuberculosis. Tuberculosis (Edinb). 2014;94:55–64.CrossRefGoogle Scholar
  29. 29.
    Dorhoi A, Kaufmann SH. Versatile myeloid cell subsets contribute to tuberculosis-associated inflammation. Eur J Immunol. 2015;45:2191–202.CrossRefPubMedGoogle Scholar
  30. 30.
    Dorhoi A, Kaufmann SH. Pathology and immune reactivity: understanding multidimensionality in pulmonary tuberculosis. Semin Immunopathol. 2016;38:153–66.CrossRefPubMedGoogle Scholar
  31. 31.
    Zimmermann M. Ethical guidelines for investigations of experimental pain in conscious animals. Pain. 1983;16:109–10.CrossRefPubMedGoogle Scholar
  32. 32.
    Henriques MG, Weg VB, Martins MA, Silva PM, Fernandes PD, Cordeiro RS, et al. Differential inhibition by two hetrazepine PAF antagonists of acute inflammation in the mouse. Br J Pharmacol. 1990;99:164–8.CrossRefPubMedPubMedCentralGoogle Scholar
  33. 33.
    Ferraris FK, Moret KH, Figueiredo AB, Penido C, Henriques M. Gedunin, a natural tetranortriterpenoid, modulates T lymphocyte responses and ameliorates allergic inflammation. Int Immunopharmacol. 2012;14:82–93.CrossRefPubMedGoogle Scholar
  34. 34.
    Werneck-Barroso E, Moura AC, Monteiro MM, de Menezes Lima O Jr, de Meirelles MN, Henriques MG. Distinct ability to accumulate eosinophils during the inflammatory cellular response to M. bovis BCG in the mouse pleural cavity. Inflamm Res. 2000;49:206–13.CrossRefPubMedGoogle Scholar
  35. 35.
    Aleman M, de la Barrera SS, Schierloh PL, Alves L, Yokobori N, Baldini M, et al. In tuberculous pleural effusions, activated neutrophils undergo apoptosis and acquire a dendritic cell-like phenotype. J Infect Dis. 2005;192:399–409.CrossRefPubMedGoogle Scholar
  36. 36.
    Afonso PV, Janka-Junttila M, Lee YJ, McCann CP, Oliver CM, Aamer KA, et al. LTB4 is a signal-relay molecule during neutrophil chemotaxis. Dev Cell. 2012;22:1079–91.CrossRefPubMedPubMedCentralGoogle Scholar
  37. 37.
    Niggli V. Signaling to migration in neutrophils: importance of localized pathways. Int J Biochem Cell Biol. 2003;35:1619–38.CrossRefPubMedGoogle Scholar
  38. 38.
    Medeiros R, Passos GF, Vitor CE, Koepp J, Mazzuco TL, Pianowski LF, et al. Effect of two active compounds obtained from the essential oil of Cordia verbenacea on the acute inflammatory responses elicited by LPS in the rat paw. Br J Pharmacol. 2007;151:618–27.CrossRefPubMedPubMedCentralGoogle Scholar
  39. 39.
    Montecucco F, Lenglet S, Braunersreuther V, Burger F, Pelli G, Bertolotto M, et al. CB(2) cannabinoid receptor activation is cardioprotective in a mouse model of ischemia/reperfusion. J Mol Cell Cardiol. 2009;46:612–20.CrossRefPubMedGoogle Scholar
  40. 40.
    Lowe DM, Redford PS, Wilkinson RJ, O’Garra A, Martineau AR. Neutrophils in tuberculosis: friend or foe? Trends Immunol. 2012;33:14–25.CrossRefPubMedGoogle Scholar
  41. 41.
    Gopal R, Monin L, Torres D, Slight S, Mehra S, McKenna KC, et al. S100A8/A9 proteins mediate neutrophilic inflammation and lung pathology during tuberculosis. Am J Respir Crit Care Med. 2013;188:1137–46.CrossRefPubMedPubMedCentralGoogle Scholar
  42. 42.
    Yeremeev V, Linge I, Kondratieva T, Apt A. Neutrophils exacerbate tuberculosis infection in genetically susceptible mice. Tuberculosis. 2015;95:447–51.CrossRefPubMedGoogle Scholar
  43. 43.
    Zhang X, Majlessi L, Deriaud E, Leclerc C, Lo-Man R. Coactivation of Syk kinase and MyD88 adaptor protein pathways by bacteria promotes regulatory properties of neutrophils. Immunity. 2009;31:761–71.CrossRefPubMedGoogle Scholar
  44. 44.
    Gui H, Liu X, Liu LR, Su DF, Dai SM. Activation of cannabinoid receptor 2 attenuates synovitis and joint distruction in collagen-induced arthritis. Immunobiology. 2015;220:817–22.CrossRefPubMedGoogle Scholar
  45. 45.
    Sawant KV, McMurray DN. Guinea pig neutrophils infected with Mycobacterium tuberculosis produce cytokines which activate alveolar macrophages in noncontact cultures. Infect Immun. 2007;75:1870–7.CrossRefPubMedPubMedCentralGoogle Scholar
  46. 46.
    Dimasi D, Sun WY, Bonder CS. Neutrophil interactions with the vascular endothelium. Int Immunopharmacol. 2013;17:1167–75.CrossRefPubMedGoogle Scholar
  47. 47.
    Rajesh M, Mukhopadhyay P, Bátkai S, Haskó G, Liaudet L, Huffman JW, et al. CB2-receptor stimulation attenuates TNF-alpha-induced human endothelial cell activation, transendothelial migration of monocytes, and monocyte-endothelial adhesion. Am J Physiol Heart Circ Physiol. 2007;293:H2210–8.CrossRefPubMedPubMedCentralGoogle Scholar
  48. 48.
    Ramirez SH, Haskó J, Skuba A, Fan S, Dykstra H, McCormick R, et al. Activation of cannabinoid receptor 2 attenuates leukocyte–endothelial cell interactions and blood–brain barrier dysfunction under inflammatory conditions. J Neurosci. 2012;32:4004–16.CrossRefPubMedPubMedCentralGoogle Scholar
  49. 49.
    Heit B, Robbins SM, Downey CM, Guan Z, Colarusso P, Miller BJ, et al. PTEN functions to ‘prioritize’ chemotactic cues and prevent ‘distraction’ in migrating neutrophils. Nat Immunol. 2008;9:743–52.CrossRefPubMedGoogle Scholar
  50. 50.
    Kurihara R, Tohyama Y, Matsusaka S, Naruse H, Kinoshita E, Tsujioka T, et al. Effects of peripheral cannabinoid receptor ligands on motility and polarization in neutrophil-like HL60 cells and human neutrophils. J Biol Chem. 2006;281:12908–18.CrossRefPubMedGoogle Scholar

Copyright information

© Springer International Publishing 2016

Authors and Affiliations

  • Magaiver Andrade-Silva
    • 1
    • 3
  • Luana Barbosa Correa
    • 1
    • 3
  • André Luis Peixoto Candéa
    • 1
    • 3
  • Simone C. Cavalher-Machado
    • 1
  • Helene Santos Barbosa
    • 2
  • Elaine Cruz Rosas
    • 1
    • 3
  • Maria G Henriques
    • 1
    • 3
    • 4
    Email author
  1. 1.Laboratório de Farmacologia Aplicada, Instituto de Tecnologia em Fármacos-FarmanguinhosFundação Oswaldo CruzRio de JaneiroBrasil
  2. 2.Laboratório de Biologia Estrutural, Instituto Oswaldo CruzFundação Oswaldo CruzRio de JaneiroBrasil
  3. 3.Instituto Nacional de Ciência e Tecnologia de Inovação em Doenças de Populações Negligenciadas (INCT-IDPN)Rio de JaneiroBrasil
  4. 4.Centro de Desenvolvimento Tecnológico em Saúde (CDTS)Rio de JaneiroBrasil

Personalised recommendations