Skip to main content
SpringerLink
Log in

Inhibition of neuroinflammation in BV2 microglia by the biflavonoid kolaviron is dependent on the Nrf2/ARE antioxidant protective mechanism

  • Published:
Molecular and Cellular Biochemistry Aims and scope Submit manuscript

Abstract

Kolaviron is a mixture of biflavonoids found in the nut of the West African edible seed Garcinia kola, and it has been reported to exhibit a wide range of pharmacological activities. In this study, we investigated the effects of kolaviron in neuroinflammation. The effects of kolaviron on the expression of nitric oxide/inducible nitric oxide synthase (iNOS), prostaglandin E2 (PGE2)/cyclooxygenase-2, cellular reactive oxygen species (ROS) and the pro-inflammatory cytokines were examined in lipopolysaccharide (LPS)-stimulated BV2 microglial cells. Molecular mechanisms of the effects of kolaviron on NF-κB and Nrf2/ARE signalling pathways were analysed by immunoblotting, binding assays and reporter assays. RNA interference was used to investigate the role of Nrf2 in the anti-inflammatory effect of kolaviron. Neuroprotective effect of kolaviron was assessed in a BV2 microglia/HT22 hippocampal neuron co-culture. Kolaviron inhibited the protein levels of NO/iNOS, PGE2/COX-2, cellular ROS and the pro-inflammatory cytokines (TNFα and IL-6) in LPS-stimulated microglia. Further mechanistic studies showed that kolaviron inhibited neuroinflammation by inhibiting IκB/NF-κB signalling pathway in LPS-activated BV2 microglia. Kolaviron produced antioxidant effect in BV2 microglia by increasing HO-1 via the Nrf2/antioxidant response element pathway. RNAi experiments revealed that Nrf2 is needed for the anti-inflammatory effects of kolaviron. Kolaviron protected HT22 neurons from neuroinflammation-induced toxicity. Kolaviron inhibits neuroinflammation through Nrf2-dependent mechanisms. This compound may therefore be beneficial in neuroinflammation-related neurodegenerative disorders.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Log in via an institution

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2
Fig. 3
Fig. 4
Fig. 5
Fig. 6
Fig. 7
Fig. 8
Fig. 9
Fig. 10
Fig. 11

Similar content being viewed by others

References

  1. Polazzi E, Monti B (2010) Microglia and neuroprotection: from in vitro studies to therapeutic applications. Prog Neurobiol 92:293–315

    Article  PubMed  Google Scholar 

  2. Li W, Khor TO, Xu C, Shen G, Jeong WK, Yu S, Kong AN (2008) Activation of Nrf2-antioxidant signaling attenuates NFκB-inflammatory response and elicits apoptosis. Biochem Pharmacol 76:1485–1489

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  3. Koh K, Kim J, Jang YJ, Yoon K, Cha Y, Lee HJ, Kim J (2011) Transcription factor Nrf2 suppresses LPS-induced hyperactivation of BV-2 microglial cells. J Neuroimmunol 233:160–167

    Article  CAS  PubMed  Google Scholar 

  4. Sandberg M, Patil J, D’Angelo B, Weber SG, Mallard C (2014) Nrf2-regulation in brain health and disease: implication of cerebral inflammation. Neuropharmacology 79:298–306

    Article  CAS  PubMed  Google Scholar 

  5. Kim J, Cha YN, Surh YJ (2010) A protective role of nuclear factor-erythroid 2-related factor-2 (Nrf2) in inflammatory disorders. Mutat Res 690:12–23

    Article  CAS  PubMed  Google Scholar 

  6. Innamorato NG, Lasters-Becker I, Cuadrado A (2009) Role of microglial redox balance in modulation of neuroinflammation. Curr Opin Neurol 22:308–314

    Article  CAS  PubMed  Google Scholar 

  7. Innamorato NG, Rojo AI, García-Yagüe AJ, Yamamoto M, de Ceballos ML, Cuadrado A (2008) The transcription factor Nrf2 is a therapeutic target against brain inflammation. J Immunol 181:680–689

    Article  CAS  PubMed  Google Scholar 

  8. Farombi EO, Tahnteng JG, Agboola AO, Nwankwo JO, Emerole GO (2000) Chemoprevention of 2-acetylaminofluorene-induced hepatotoxicity and lipid peroxidation in rats by kolaviron-a Garcinia kola seed extract. Food Chem Toxicol 38:535–541

    Article  CAS  PubMed  Google Scholar 

  9. Farombi EO (2000) Mechanisms for the hepatoprotective action of kolaviron: studies on hepatic enzymes, microsomal lipids and lipid peroxidation in carbon tetrachloride-treated rats. Pharmacol Res 42:75–80

    Article  CAS  PubMed  Google Scholar 

  10. Nwankwo JO, Tahnteng JG, Emerole GO (2000) Inhibition of aflatoxin B1 genotoxicity in human liver-derived HepG2 cells by kolaviron biflavonoids and molecular mechanisms of action. Eur J Cancer Prev 9:351–361

    Article  CAS  PubMed  Google Scholar 

  11. Abarikwu SO, Farombi EO, Pant AB (2011) Biflavanone-kolaviron protects human dopaminergic SH-SY5Y cells against atrazine induced toxic insult. Toxicol In Vitro 25:848–858

    Article  CAS  PubMed  Google Scholar 

  12. Igado OO, Olopade JO, Adesida A, Aina OO, Farombi EO (2012) Morphological and biochemical investigation into the possible neuroprotective effects of kolaviron (Garcinia kola bioflavonoid) on the brains of rats exposed to vanadium. Drug Chem Toxicol 35:371–380

    Article  CAS  PubMed  Google Scholar 

  13. Olajide OJ, Enaibe BU, Bankole OO, Akinola OB, Laoye BJ, Ogundele OM (2015) Kolaviron was protective against sodium azide (NaN3) induced oxidative stress in the prefrontal cortex. Metab Brain Dis 31:25–35

    Article  PubMed  Google Scholar 

  14. Olaleye SB, Onasanwo SA, Ige AO, Wu KK, Cho CH (2010) Anti-inflammatory activities of a kolaviron-inhibition of nitric oxide, prostaglandin E2 and tumor necrosis factor-alpha production in activated macrophage-like cell line. Afr J Med Med Sci 39:41–46

    PubMed  Google Scholar 

  15. Abarikwu SO (2014) Kolaviron, a natural flavonoid from the seeds of Garcinia kola, reduces LPS-induced inflammation in macrophages by combined inhibition of IL-6 secretion, and inflammatory transcription factors, ERK1/2, NF-κB, p38, Akt, p-c-JUN and JNK. Biochim Biophys Acta 1840:2373–2381

    Article  CAS  PubMed  Google Scholar 

  16. Farombi EO, Shrotriya S, Surh YJ (2009) Kolaviron inhibits dimethyl nitrosamine-induced liver injury by suppressing COX-2 and iNOS expression via NF-kappaB and AP-1. Life Sci 84(2009):149–155

    Article  CAS  PubMed  Google Scholar 

  17. Iwu MM (1985) Antihepatoxic constituents of Garcinia kola seeds. Experientia 41:699–700

    Article  CAS  PubMed  Google Scholar 

  18. Iwu MM, Igboko OA, Okunji CO, Tempesta MS (1990) Antidiabetic and aldose reductase activities of biflavanones of Garcinia kola. J Pharm Pharmacol 42:290–292

    Article  CAS  PubMed  Google Scholar 

  19. Okorji UP, Olajide OA (2014) A semi-synthetic derivative of artemisinin, artesunate inhibits prostaglandin E2 production in LPS/IFNγ-activated BV2 microglia. Bioorg Med Chem. 22:4726–4734

    Article  CAS  PubMed  Google Scholar 

  20. Fiebich BL, Akter S, Akundi RS (2014) The two-hit hypothesis for neuroinflammation: role of exogenous ATP in modulating inflammation in the brain. Front Cell Neurosci 8:260

    Article  PubMed  PubMed Central  Google Scholar 

  21. Olajide OA, Kumar A, Velagapudi R, Okorji UP, Fiebich BL (2014) Punicalagin inhibits neuroinflammation in LPS-activated rat primary microglia. Mol Nutr Food Res 58:1843–1851

    Article  CAS  PubMed  Google Scholar 

  22. Dumont M, Beal MF (2011) Neuroprotective strategies involving ROS in Alzheimer disease. Free Radic Biol Med 51:1014–1026

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  23. Gao HM, Zhou H, Hong JS (2012) NADPH oxidases: novel therapeutic targets for neurodegenerative diseases. Trends Pharmacol Sci 33:295–303

    Article  PubMed  PubMed Central  Google Scholar 

  24. Syapin PJ (2008) Regulation of haeme oxygenase-1 for treatment of neuroinflammation and brain disorders. Br J Pharmacol 155:623–640

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  25. Yuste JE, Tarragon E, Campuzano CM, Ros-Bernal F (2015) Implications of glial nitric oxide in neurodegenerative diseases. Front Cell Neurosci 9:322

    Article  PubMed  PubMed Central  Google Scholar 

  26. Miyagishi H, Kosuge Y, Yoneoka Y, Ozone M, Endo M, Osada N, Ishige K, Kusama-Eguchi K, Ito Y (2013) Prostaglandin E2-induced cell death is mediated by activation of EP2 receptors in motor neuron-like NSC-34 cells. J Pharmacol Sci 121:347–350

    Article  CAS  PubMed  Google Scholar 

  27. Block ML, Hong JS (2005) Microglia and inflammation-mediated neurodegeneration: multiple triggers with a common mechanism. Prog Neurobiol 76:77–98

    Article  CAS  PubMed  Google Scholar 

  28. Abarikwu SO (2015) Anti-inflammatory effects of kolaviron modulate the expressions of inflammatory marker genes, inhibit transcription factors ERK1/2, p-JNK, NF-κB, and activate Akt expressions in the 93RS2 Sertoli cell lines. Mol Cell Biochem 401:197–208

    Article  CAS  PubMed  Google Scholar 

  29. Ratajczak-Wrona W, Jablonska E, Garley M, Jablonski J, Radziwon P, Iwaniuk A, Grubczak K (2014) PI3 K-Akt/PKB signaling pathway in neutrophils and mononuclear cells exposed to N-nitrosodimethylamine. J Immunotoxicol 11:231–237

    Article  CAS  PubMed  Google Scholar 

  30. Lee S, Suk K (2007) Heme oxygenase-1 mediates cytoprotective effects of immunostimulation in microglia. Biochem Pharmacol 74:723–729

    Article  CAS  PubMed  Google Scholar 

  31. Velagapudi R, Aderogba M, Olajide OA (2014) Tiliroside, a dietary glycosidic flavonoid, inhibits TRAF-6/NF-κB/p38-mediated neuroinflammation in activated BV2 microglia. Biochim Biophys Acta 1840:3311–3319

    Article  CAS  PubMed  Google Scholar 

  32. Foresti R, Bains SK, Pitchumony TS, de Castro Brás LE, Drago F, Dubois-Randé JL, Bucolo C, Motterlini R (2013) Small molecule activators of the Nrf2-HO-1 antioxidant axis modulate heme metabolism and inflammation in BV2 microglia cells. Pharmacol Res 76:132–148

    Article  CAS  PubMed  Google Scholar 

  33. Zujovic V, Taupin V (2003) Use of cocultured cell systems to elucidate chemokine-dependent neuronal/microglial interactions: control of microglial activation. Methods 29:345–350

    Article  CAS  PubMed  Google Scholar 

Download references

Acknowledgments

The authors would like to thank the Physiological Society for funding this study, and for the award of an International Junior Research Grant (IJRG) to Dr Samuel A. Onasanwo.

Authors’ contributions

OAO designed the study and wrote the manuscript. SAO, RV, AE and OAO performed the experiments. OAO and RV analysed the data. All authors read and approved the final manuscript.

Author information

Author notes

    Authors and Affiliations

    1. Department of Physiology, College of Medicine, University of Ibadan, Ibadan, Nigeria

      Samuel A. Onasanwo

    2. Department of Pharmacy, School of Applied Sciences, University of Huddersfield, Queensgate, Huddersfield, HD1 3DH, UK

      Ravikanth Velagapudi, Abdelmeneim El-Bakoush & Olumayokun A. Olajide

    Authors
    1. Samuel A. Onasanwo
      View author publications

      You can also search for this author in PubMed Google Scholar

    2. Ravikanth Velagapudi
      View author publications

      You can also search for this author in PubMed Google Scholar

    3. Abdelmeneim El-Bakoush
      View author publications

      You can also search for this author in PubMed Google Scholar

    4. Olumayokun A. Olajide
      View author publications

      You can also search for this author in PubMed Google Scholar

    Corresponding author

    Correspondence to Olumayokun A. Olajide.

    Ethics declarations

    Conflict of Interest

    The authors declare that they have no competing interests.

    Additional information

    Samuel A. Onasanwo and Ravikanth Velagapudi have contributed equally to this work.

    Rights and permissions

    Reprints and permissions

    About this article

    Check for updates. Verify currency and authenticity via CrossMark

    Cite this article

    Onasanwo, S.A., Velagapudi, R., El-Bakoush, A. et al. Inhibition of neuroinflammation in BV2 microglia by the biflavonoid kolaviron is dependent on the Nrf2/ARE antioxidant protective mechanism. Mol Cell Biochem 414, 23–36 (2016). https://doi.org/10.1007/s11010-016-2655-8

    Download citation

    • Received:

    • Accepted:

    • Published:

    • Issue Date:

    • DOI: https://doi.org/10.1007/s11010-016-2655-8

    Keywords

    Search

    Navigation