Cannabinoid receptor 2 activation mitigates lipopolysaccharide-induced neuroinflammation and sickness behavior in mice

  • Puspita Sahu
  • Jayesh Mudgal
  • Devinder Arora
  • Manas Kinra
  • Sanchari Basu Mallik
  • Chamallamudi Mallikarjuna Rao
  • K. S. R. Pai
  • Madhavan NampoothiriEmail author
Original Investigation


Rationale and objectives

Cannabinoid receptor 2 (CB2R) signaling in the brain is associated with the pathophysiology of depression. Sickness behavior, characterized by lessened mobility, social interaction, and depressive behavior, is linked with neuroinflammation, oxidative stress, and immune system. The present study was aimed at evaluating 1-phenylisatin (PI), a CB2R agonist, in sickness behavior.


Influence of acute and 7-day activation of CB2R using PI in lipopolysaccharide (LPS)-induced sickness behavior was assessed in mice. An acute injection of LPS (1.5 mg/kg) produced a fully developed sickness behavior in animals within 1 h of administration. The behavioral paradigm was assessed by open field test, forced swim test, and tail suspension test. Further, tumor necrosis factor-α (TNF-α), antioxidant enzymes, and lipid peroxidation were measured in the brain to correlate neuroinflammation and oxidative stress with sickness behavior. Both treatments, PI (20 mg/kg) and imipramine (15 mg/kg), were administered orally (once for acute and once daily for 7-day protocols).


LPS elevated the brain TNF-α level, augmented oxidative stress, and induced the sickness behavior in mice. Acute and 7-day treatment of mice with PI significantly reduced the LPS-induced sickness behavior. In addition, PI inhibited the neuroinflammation evidenced by a reduction in brain TNF-α and oxidative stress.


Our data propose that acute and long-term activation of CB2R might prevent neuroinflammation and oxidative stress-associated sickness behavior.


Cannabinoid Phenylisatin Sickness behavior Neuroinflammation Oxidative stress Depression 



The authors thank the Manipal Academy of Higher Education, Manipal, for providing the research facilities.

Compliance with ethical standards

Institutional Animal Ethics Committee approved the experimental protocol. The experiments were performed following the Committee for the Purpose of Control and Supervision of Experiments on Animals (CPCSEA) guidelines.

Conflict of interest

The authors declare that they have no conflict of interest.


  1. Aebi H (1984) Catalase in vitro. Methods Enzymol 105:121–126. CrossRefPubMedPubMedCentralGoogle Scholar
  2. Benito C, Tolon RM, Pazos MR, Nunez E, Castillo AI, Romero J (2008) Cannabinoid CB2 receptors in human brain inflammation. Br J Pharmacol 153(2):277–285. CrossRefPubMedGoogle Scholar
  3. Bhattacharya SK, Bhattacharya A, Kumar A, Ghosal S (2000) Antioxidant activity of Bacopa monniera in rat frontal cortex, striatum and hippocampus. Phytother Res 14(3):174–179Google Scholar
  4. Bluthe RM, Laye S, Michaud B, Combe C, Dantzer R, Parnet P (2000) Role of interleukin-1β and tumour necrosis factor-α in lipopolysaccharide-induced sickness behaviour: a study with interleukin-1 type I receptor-deficient mice. Eur J Neurosci 12(12):4447–4456. CrossRefPubMedGoogle Scholar
  5. Bradford MM (1976) A rapid and sensitive method for the quantitation of microgram quantities of protein utilizing the principle of protein-dye binding. Anal Biochem 72(1–2):248–254. CrossRefPubMedPubMedCentralGoogle Scholar
  6. Buch SJ (2013) Cannabinoid receptor 2 activation: a means to prevent monocyte–endothelium engagement. Am J Pathol 183(5):1375–1377CrossRefGoogle Scholar
  7. Chung YC, Shin WH, Baek JY, Cho EJ, Baik HH, Kim SR, Won SY, Jin BK (2016) CB2 receptor activation prevents glial-derived neurotoxic mediator production, BBB leakage and peripheral immune cell infiltration and rescues dopamine neurons in the MPTP model of Parkinson’s disease. Exp Mol Med 48(1):e205. CrossRefPubMedPubMedCentralGoogle Scholar
  8. García-Gutierrez MS, Manzanares J (2011) Overexpression of CB2 cannabinoid receptors decreased vulnerability to anxiety and impaired anxiolytic action of alprazolam in mice. J Psychopharmacol 25(1):111–120. CrossRefPubMedGoogle Scholar
  9. Howlett AC (2005) Cannabinoid receptor signaling. Handb Exp Pharmacol 168:53–79. CrossRefGoogle Scholar
  10. Janero DR (1990) Malondialdehyde and thiobarbituric acid-reactivity as diagnostic indices of lipid peroxidation and peroxidative tissue injury. Free Radic Biol Med 9(6):515–540. CrossRefGoogle Scholar
  11. Jayant S, Sharma BM, Bansal R, Sharma B (2016) Pharmacological benefits of selective modulation of cannabinoid receptor type 2 (CB2) in experimental Alzheimer’s disease. Pharmacol Biochem Behav 140:39–50. CrossRefPubMedGoogle Scholar
  12. Justo LA, Durán R, Alfonso M, Fajardo D, Faro LR (2016) Effects and mechanism of action of isatin, a MAO inhibitor, on in vivo striatal dopamine release. Neurochem Int 99:147–157. CrossRefPubMedGoogle Scholar
  13. Kruk-Slomka M, Banaszkiewicz I, Biala G (2017) The impact of CB2 receptor ligands on the MK-801-induced hyperactivity in mice. Neurotox Res 31(3):410–420Google Scholar
  14. Kumar N, Dhayabaran D, Nampoothiri M, Nandakumar K, Puratchikody A, Lalani N, Dawood K, Ghosh A (2014). Atypical antidepressant activity of 3,4-Bis (3,4-dimethoxyphenyl) furan-2,5-dione isolated from heart wood of cedrus deodara, in rodents. The Korean Journal of Physiology & Pharmacology 18(5):365–369Google Scholar
  15. Kurosawa N, Shimizu K, Seki K (2016) The development of depression-like behavior is consolidated by IL-6-induced activation of locus coeruleus neurons and IL-1β-induced elevated leptin levels in mice. Psychopharmacol 233(9):1725–1737. CrossRefGoogle Scholar
  16. Liu QR, Canseco-Alba A, Zhang HY, Tagliaferro P, Chung M, Dennis E, Sanabria B, Schanz N, Escosteguy-Neto JC, Ishiguro H, Lin Z (2017) Cannabinoid type 2 receptors in dopamine neurons inhibits psychomotor behaviors, alters anxiety, depression and alcohol preference. Sci Rep 7(1):17410. CrossRefPubMedPubMedCentralGoogle Scholar
  17. Lou ZY, Chen C, He Q, Zhao CB, Xiao BG (2011) Targeting CB2 receptor as a neuroinflammatory modulator in experimental autoimmune encephalomyelitis. Mol Immunol 49(3):453–461. CrossRefPubMedGoogle Scholar
  18. Mallik SB, Mudgal J, Nampoothiri M, Hall S, Anoopkumar-Dukie S, Grant G, Rao CM, Arora D (2016) Caffeic acid attenuates lipopolysaccharide-induced sickness behaviour and neuroinflammation in mice. Neurosci Lett 632:218–223. CrossRefGoogle Scholar
  19. Moron MS, Depierre JW, Mannervik B (1979) Levels of glutathione, glutathione reductase and glutathione S-transferase activities in rat lung and liver. Biochim Biophys Acta 582(1):67–78.
  20. Nestler EJ, Carlezon WA (2006) The mesolimbic dopamine reward circuit in depression. Biol Psychiatry 59(12):1151–1159. CrossRefPubMedGoogle Scholar
  21. O’connor JC, Lawson MA, Andre C, Moreau M, Lestage J, Castanon N, Kelley KW, Dantzer R (2009) Lipopolysaccharide-induced depressive-like behavior is mediated by indoleamine 2, 3-dioxygenase activation in mice. Mol Psychiatry 14(5):511–522. CrossRefPubMedGoogle Scholar
  22. Painsipp E, Kofer MJ, Sinner F, Holzer P (2011) Prolonged depression-like behavior caused by immune challenge: influence of mouse strain and social environment. PLoS One 6(6):e20719. CrossRefPubMedPubMedCentralGoogle Scholar
  23. Perdikaris P, Tsarouchi M, Fanarioti E, Natsaridis E, Mitsacos A, Giompres P (2018) Long lasting effects of chronic WIN55, 212-2 treatment on mesostriatal dopaminergic and cannabinoid systems in the rat brain. Neuropharmacol 129:1–5. CrossRefGoogle Scholar
  24. Pertwee RG, Howlett AC, Abood ME, Alexander SPH, Di Marzo V, Elphick MR, Greasley PJ, Hansen HS, Kunos G, Mackie K, Mechoulam R (2010) International Union of Basic and Clinical Pharmacology. LXXIX. Cannabinoid receptors and their ligands: beyond CB1 and CB2. Pharmacol Rev 62:588–631. CrossRefPubMedPubMedCentralGoogle Scholar
  25. Porsolt RD, Bertin A, Jalfre M (1978) “Behavioural despair” in rats and mice: strain differences and the effects of imipramine. Eur J Pharmacol 51(3):291–294. CrossRefPubMedGoogle Scholar
  26. Ren Z, Yan P, Zhu L, Yang H, Zhao Y, Kirby BP, Waddington JL, Zhen X (2018) Dihydromyricetin exerts a rapid antidepressant-like effect in association with enhancement of BDNF expression and inhibition of neuroinflammation. Psychopharmacol 235(1):233–244. CrossRefGoogle Scholar
  27. Tambaro S, Casu MA, Mastinu A, Lazzari P (2014) Evaluation of selective cannabinoid CB1 and CB2 receptor agonists in a mouse model of lipopolysaccharide-induced interstitial cystitis. Eur J Pharmacol 729:67–74. CrossRefPubMedGoogle Scholar
  28. Teixeira RC, Zangrossi H Jr, Graeff FG (2000) Behavioral effects of acute and chronic imipramine in the elevated T-maze model of anxiety. Pharmacol Biochem Behav 65(4):571–576. CrossRefPubMedGoogle Scholar
  29. Tye KM, Mirzabekov JJ, Warden MR, Ferenczi EA, Tsai HC, Finkelstein J, Kim SY, Adhikari A, Thompson KR, Andalman AS, Gunaydin LA (2013) Dopamine neurons modulate neural encoding and expression of depression-related behaviour. Nature 493(7433):537–541. CrossRefPubMedGoogle Scholar
  30. Varatharaj A, Galea I (2017) The blood-brain barrier in systemic inflammation. Brain Behav Immun 60:1–2. CrossRefPubMedGoogle Scholar
  31. Wu J, Hocevar M, Foss JF, Bie B, Naguib M (2017) Activation of CB2 receptor system restores cognitive capacity and hippocampal Sox2 expression in a transgenic mouse model of Alzheimer’s disease. Eur J Pharmacol 811:12–20. CrossRefPubMedGoogle Scholar
  32. Yirmiya R (1996) Endotoxin produces a depressive-like episode in rats. Brain Res 711(1–2):163–174. CrossRefPubMedGoogle Scholar

Copyright information

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

Authors and Affiliations

  1. 1.Department of Pharmacology, Manipal College of Pharmaceutical SciencesManipal Academy of Higher EducationManipalIndia
  2. 2.School of Pharmacy and Pharmacology, MHIQ, QUM NetworkGriffith UniversityGold CoastAustralia

Personalised recommendations