Naringenin is a biologically active analgesic, anti-inflammatory, and antioxidant flavonoid. Naringenin targets in inflammation-induced articular pain remain poorly explored.
The present study investigated the cellular and molecular mechanisms involved in the analgesic/anti-inflammatory effects of naringenin in zymosan-induced arthritis. Mice were pre-treated orally with naringenin (16.7–150 mg/kg), followed by intra-articular injection of zymosan. Articular mechanical hyperalgesia and oedema, leucocyte recruitment to synovial cavity, histopathology, expression/production of pro- and anti-inflammatory mediators and NFκB activation, inflammasome component expression, and oxidative stress were evaluated.
Naringenin inhibited articular pain and oedema in a dose-dependent manner. The dose of 50 mg/kg inhibited leucocyte recruitment, histopathological alterations, NFκB activation, and NFκB-dependent pro-inflammatory cytokines (TNF-α, IL-1β, and IL-33), and preproET-1 mRNA expression, but increased anti-inflammatory IL-10. Naringenin also inhibited inflammasome upregulation (reduced Nlrp3, ASC, caspase-1, and pro-IL-1β mRNA expression) and oxidative stress (reduced gp91phox mRNA expression and superoxide anion production, increased GSH levels, induced Nrf2 protein in CD45+ hematopoietic recruited cells, and induced Nrf2 and HO-1 mRNA expression).
Naringenin presents analgesic and anti-inflammatory effects in zymosan-induced arthritis by targeting its main physiopathological mechanisms. These data highlight this flavonoid as an interesting therapeutic compound to treat joint inflammation, deserving additional pre-clinical and clinical studies.
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Aletaha D, Smolen JS (2018) Diagnosis and management of rheumatoid arthritis: a review. JAMA 320:1360–1372. https://doi.org/10.1001/jama.2018.13103
Ali R, Shahid A, Ali N, Hasan SK, Majed F, Sultana S (2016) Amelioration of benzo[a]pyrene-induced oxidative stress and pulmonary toxicity by Naringenin in Wistar rats: a plausible role of COX-2 and NF-kappaB. Hum Exp Toxicol. https://doi.org/10.1177/0960327116650009
Al-Rejaie SS, Aleisa AM, Abuohashish HM, Parmar MY, Ola MS, Al-Hosaini AA, Ahmed MM (2015) Naringenin neutralises oxidative stress and nerve growth factor discrepancy in experimental diabetic neuropathy. Neurol Res 37:924–933. https://doi.org/10.1179/1743132815Y.0000000079
Ananth DA, Rameshkumar A, Jeyadevi R, Aseervatham GS, Sripriya J, Bose PC, Sivasudha T (2016) Amelioratory effect of flavonoids rich Pergularia daemia extract against CFA induced arthritic rats. Biomed Pharmacothery = Biomedecine and pharmacotherapie 80:244–252. https://doi.org/10.1016/j.biopha.2016.03.019
Aruoma OI (2003) Methodological considerations for characterizing potential antioxidant actions of bioactive components in plant foods. Mutat Res 523–524:9–20
Asquith DL, Miller AM, McInnes IB, Liew FY (2009) Animal models of rheumatoid arthritis. Eur J Immunol 39:2040–2044. https://doi.org/10.1002/eji.200939578
Bai X et al (2014) Protective effect of naringenin in experimental ischemic stroke: down-regulated NOD2, RIP2, NF-kappaB, MMP-9 and up-regulated claudin-5 expression. Neurochem Res 39:1405–1415. https://doi.org/10.1007/s11064-014-1326-y
Bernardy CCF et al (2017) Tempol, a superoxide dismutase mimetic agent, inhibits superoxide anion-induced inflammatory pain in mice. BioMed Res Int 2017:9584819. https://doi.org/10.1155/2017/9584819
Bouayed J, Bohn T (2010) Exogenous antioxidants—double-edged swords in cellular redox state: Health beneficial effects at physiologic doses versus deleterious effects at high doses. Oxid Med Cell Longev 3:228–237. https://doi.org/10.4161/oxim.3.4.12858
Chtourou Y, Fetoui H, Jemai R, Ben Slima A, Makni M, Gdoura R (2015) Naringenin reduces cholesterol-induced hepatic inflammation in rats by modulating matrix metalloproteinases-2, 9 via inhibition of nuclear factor kappaB pathway. Eur J Pharmacol 746:96–105. https://doi.org/10.1016/j.ejphar.2014.10.027
Conte Fde P, Barja-Fidalgo C, Verri WA Jr, Cunha FQ, Rae GA, Penido C, Henriques M (2008) Endothelins modulate inflammatory reaction in zymosan-induced arthritis: participation of LTB4, TNF-alpha, and CXCL-1. J Leukoc Biol 84:652–660. https://doi.org/10.1189/jlb.1207827
Conte FP, Menezes-de-Lima O Jr, Verri WA Jr, Cunha FQ, Penido C, Henriques MG (2010) Lipoxin A(4) attenuates zymosan-induced arthritis by modulating endothelin-1 and its effects. Br J Pharmacol 161:911–924. https://doi.org/10.1111/j.1476-5381.2010.00950.x
Craig W, Poppema S, Little MT, Dragowska W, Lansdorp PM (1994) CD45 isoform expression on human haemopoietic cells at different stages of development. Br J Haematol 88:24–30
Cross M et al (2014) The global burden of rheumatoid arthritis: estimates from the global burden of disease 2010 study. Ann Rheum Dis 73:1316–1322. https://doi.org/10.1136/annrheumdis-2013-204627
Decker EA (1997) Phenolics: prooxidants or antioxidants? Nutr Rev 55:396–398
Donate PB et al (2012) Bosentan, an endothelin receptor antagonist, ameliorates collagen-induced arthritis: the role of TNF-alpha in the induction of endothelin system genes. Inflamm Res 61:337–348. https://doi.org/10.1007/s00011-011-0415-5
Du Z, Kelly E, Mecklenbrauker I, Agle L, Herrero C, Paik P, Ivashkiv LB (2006) Selective regulation of IL-10 signaling and function by zymosan. J Immunol 176:4785–4792
Esmaeili MA, Alilou M (2014) Naringenin attenuates CCl4-induced hepatic inflammation by the activation of an Nrf2-mediated pathway in rats. Clin Exp Pharmacol Physiol 41:416–422. https://doi.org/10.1111/1440-1681.12230
Fattori V et al (2016) Differential regulation of oxidative stress and cytokine production by endothelin ETA and ETB receptors in superoxide anion-induced inflammation and pain in mice. J Drug Target 15:1–27. https://doi.org/10.1080/1061186x.2016.1245308
Frabasile S, Koishi AC, Kuczera D, Silveira GF, Verri WA Jr, Duarte Dos Santos CN, Bordignon J (2017) The citrus flavanone naringenin impairs dengue virus replication in human cells. Sci Rep 7:41864. https://doi.org/10.1038/srep41864
Guerrero AT et al (2006) Hypernociception elicited by tibio-tarsal joint flexion in mice: a novel experimental arthritis model for pharmacological screening. Pharmacol Biochem Behav 84:244–251. https://doi.org/10.1016/j.pbb.2006.05.008
Guerrero AT, Cunha TM, Verri WA Jr, Gazzinelli RT, Teixeira MM, Cunha FQ, Ferreira SH (2012) Toll-like receptor 2/MyD88 signaling mediates zymosan-induced joint hypernociception in mice: participation of TNF-alpha, IL-1beta and CXCL1/KC. Eur J Pharmacol 674:51–57. https://doi.org/10.1016/j.ejphar.2011.10.023
Hu CY, Zhao YT (2014) Analgesic effects of naringenin in rats with spinal nerve ligation-induced neuropathic pain. Biomed Rep 2:569–573. https://doi.org/10.3892/br.2014.267
Jin L, Zeng W, Zhang F, Zhang C, Liang W (2017) Naringenin ameliorates acute inflammation by regulating intracellular cytokine degradation. J Immunol 199:3466–3477. https://doi.org/10.4049/jimmunol.1602016
Kaulaskar S, Bhutada P, Rahigude A, Jain D, Harle U (2012) Effects of naringenin on allodynia and hyperalgesia in rats with chronic constriction injury-induced neuropathic pain. Zhong Xi Yi Jie He Xue Bao 10:1482–1489. https://doi.org/10.3736/jcim20121223
Kesselheim AS, Avorn J, Sarpatwari A (2016) The high cost of prescription drugs in the United States: origins and prospects for reform. JAMA 316:858–871. https://doi.org/10.1001/jama.2016.11237
Kongpichitchoke T, Hsu JL, Huang TC (2015) Number of hydroxyl groups on the B-ring of flavonoids affects their antioxidant activity and interaction with phorbol ester binding site of PKCdelta C1B domain: vitro and in silico studies. J Agric Food Chem 63:4580–4586. https://doi.org/10.1021/acs.jafc.5b00312
Krogholm KS, Bredsdorff L, Knuthsen P, Haraldsdottir J, Rasmussen SE (2010) Relative bioavailability of the flavonoids quercetin, hesperetin and naringenin given simultaneously through diet. Eur J Clin Nutr 64:432–435. https://doi.org/10.1038/ejcn.2010.6
Kumar S, Pandey AK (2013) Chemistry and biological activities of flavonoids: an overview. Sci World J 2013:162750. https://doi.org/10.1155/2013/162750
Laev SS, Salakhutdinov NF (2015) Anti-arthritic agents: progress and potential. Bioorganic Med Chem 23:3059–3080. https://doi.org/10.1016/j.bmc.2015.05.010
Lamkanfi M, Malireddi RK, Kanneganti TD (2009) Fungal zymosan and mannan activate the cryopyrin inflammasome. J Biol Chem 284:20574–20581. https://doi.org/10.1074/jbc.M109.023689
Li YR, Chen DY, Chu CL, Li S, Chen YK, Wu CL, Lin CC (2015) Naringenin inhibits dendritic cell maturation and has therapeutic effects in a murine model of collagen-induced arthritis. J Nutr Biochem 26:1467–1478. https://doi.org/10.1016/j.jnutbio.2015.07.016
Lim W, Park S, Bazer FW, Song G (2017) Naringenin-induced apoptotic cell death in prostate cancer cells is mediated via the PI3K/AKT and MAPK signaling pathways. J Cell Biochem 118:1118–1131. https://doi.org/10.1002/jcb.25729
Liu K et al (2016) Chemical evidence for potent xanthine oxidase inhibitory activity of ethyl acetate extract of citrus aurantium L. Dried Immature Fruits Mol 21:302. https://doi.org/10.3390/molecules21030302
Manach C, Morand C, Gil-Izquierdo A, Bouteloup-Demange C, Remesy C (2003) Bioavailability in humans of the flavanones hesperidin and narirutin after the ingestion of two doses of orange juice. Eur J Clin Nutr 57:235–242. https://doi.org/10.1038/sj.ejcn.1601547
Manchope MF et al (2016) Naringenin inhibits superoxide anion-induced inflammatory pain: role of oxidative stress, cytokines, Nrf-2 and the NO-cGMP-PKG-KATP channel signaling pathway. PLoS One 11:e0153015. https://doi.org/10.1371/journal.pone.0153015
Manchope MF, Casagrande R, Verri WA Jr (2017) Naringenin an analgesic and anti-inflammatory citrus flavanone. Oncotarget 8:3766–3767. https://doi.org/10.18632/oncotarget.14084
Martinez RM et al (2015) Naringenin inhibits UVB irradiation-induced inflammation and oxidative stress in the skin of hairless mice. J Nat Prod 78:1647–1655. https://doi.org/10.1021/acs.jnatprod.5b00198
Nahmias Y, Goldwasser J, Casali M, van Poll D, Wakita T, Chung RT, Yarmush ML (2008) Apolipoprotein B-dependent hepatitis C virus secretion is inhibited by the grapefruit flavonoid naringenin. Hepatology 47:1437–1445. https://doi.org/10.1002/hep.22197
Naito Y, Takagi T, Higashimura Y (2014) Heme oxygenase-1 and anti-inflammatory M2 macrophages. Arch Biochem Biophys 564:83–88. https://doi.org/10.1016/j.abb.2014.09.005
Oeckinghaus A, Ghosh S (2009) The NF-kappaB family of transcription factors and its regulation. Cold Spring Harb Perspect Biol 1:a000034. https://doi.org/10.1101/cshperspect.a000034
Park HJ, Choi YJ, Lee JH, Nam MJ (2017) Naringenin causes ASK1-induced apoptosis via reactive oxygen species in human pancreatic cancer cells. Food Chem Toxicol 99:1–8. https://doi.org/10.1016/j.fct.2016.11.008
Pinho-Ribeiro FA, Zarpelon AC, Fattori V, Manchope MF, Mizokami SS, Casagrande R, Verri WA Jr (2016a) Naringenin reduces inflammatory pain in mice. Neuropharmacology 105:508–519. https://doi.org/10.1016/j.neuropharm.2016.02.019
Pinho-Ribeiro FA et al (2016b) The citrus flavonone naringenin reduces lipopolysaccharide-induced inflammatory pain and leukocyte recruitment by inhibiting NF-kappaB activation. J Nutr Biochem 33:8–14. https://doi.org/10.1016/j.jnutbio.2016.03.013
Ramprasath T, Senthamizharasi M, Vasudevan V, Sasikumar S, Yuvaraj S, Selvam GS (2014) Naringenin confers protection against oxidative stress through upregulation of Nrf2 target genes in cardiomyoblast cells. J Physiol Biochem 70:407–415. https://doi.org/10.1007/s13105-014-0318-3
Raza H, John A (2005) Green tea polyphenol epigallocatechin-3-gallate differentially modulates oxidative stress in PC12 cell compartments. Toxicol Appl Pharmacol 207:212–220. https://doi.org/10.1016/j.taap.2005.01.004
Ruiz-Miyazawa KW et al (2018) The citrus flavanone naringenin reduces gout-induced joint pain and inflammation in mice by inhibiting the activation of NFκB and macrophage release of IL-1β. J Funct Foods 48:106–116. https://doi.org/10.1016/j.jff.2018.06.025
Sahu SC, Gray GC (1997) Lipid peroxidation and DNA damage induced by morin and naringenin in isolated rat liver nuclei. Food Chem Toxicol 35:443–447
Salvemini D, Little JW, Doyle T, Neumann WL (2011) Roles of reactive oxygen and nitrogen species in pain. Free Radic Biol Med 51:951–966. https://doi.org/10.1016/j.freeradbiomed.2011.01.026
Schaible HG (2014) Nociceptive neurons detect cytokines in arthritis. Arthritis Res Ther 16:470
Schroder K, Tschopp J (2010) The inflammasomes. Cell 140:821–832. https://doi.org/10.1016/j.cell.2010.01.040
Shulman M et al (2011) Enhancement of naringenin bioavailability by complexation with hydroxypropyl-beta-cyclodextrin [corrected]. PLoS One 6:e18033. https://doi.org/10.1371/journal.pone.0018033
Souza GR et al (2015) Involvement of nuclear factor kappa B in the maintenance of persistent inflammatory hypernociception. Pharmacol Biochem Behav 134:49–56. https://doi.org/10.1016/j.pbb.2015.04.005
Sun H et al (2013) Pharmacokinetics of hesperetin and naringenin in the Zhi Zhu Wan, a traditional Chinese medicinal formulae, and its pharmacodynamics study. Phytother Res: PTR 27:1345–1351. https://doi.org/10.1002/ptr.4867
Teixeira JM, Bobinski F, Parada CA, Sluka KA, Tambeli CH (2016) P2X3 and P2X2/3 receptors play a crucial role in articular hyperalgesia development through inflammatory mechanisms in the knee joint experimental synovitis. Mol Neurobiol 15:45. https://doi.org/10.1007/s12035-016-0146-2
Vafeiadou K, Vauzour D, Lee HY, Rodriguez-Mateos A, Williams RJ, Spencer JP (2009) The citrus flavanone naringenin inhibits inflammatory signalling in glial cells and protects against neuroinflammatory injury. Arch Biochem Biophys 484:100–109. https://doi.org/10.1016/j.abb.2009.01.016
Verri WA Jr, Cunha TM, Parada CA, Poole S, Cunha FQ, Ferreira SH (2006) Hypernociceptive role of cytokines and chemokines: targets for analgesic drug development? Pharmacol Ther 112:116–138. https://doi.org/10.1016/j.pharmthera.2006.04.001
Verri WA Jr et al (2008) IL-33 mediates antigen-induced cutaneous and articular hypernociception in mice. Proc Natl Acad Sci USA 105:2723–2728. https://doi.org/10.1073/pnas.0712116105
Verri WA Jr et al (2010) IL-33 induces neutrophil migration in rheumatoid arthritis and is a target of anti-TNF therapy. Ann Rheum Dis 69:1697–1703. https://doi.org/10.1136/ard.2009.122655
Wang W et al (2014) The inhibition of RANKL-induced osteoclastogenesis through the suppression of p38 signaling pathway by naringenin and attenuation of titanium-particle-induced osteolysis. Int J Mol Sci 15:21913–21934. https://doi.org/10.3390/ijms151221913
Watjen W et al (2005) Low concentrations of flavonoids are protective in rat H4IIE cells whereas high concentrations cause DNA damage and apoptosis. J Nutr 135:525–531. https://doi.org/10.1093/jn/135.3.525
Yen HR, Liu CJ, Yeh CC (2015) Naringenin suppresses TPA-induced tumor invasion by suppressing multiple signal transduction pathways in human hepatocellular carcinoma cells. Chem Biol Interact 235:1–9. https://doi.org/10.1016/j.cbi.2015.04.003
Zarpelon AC et al (2016) Spinal cord oligodendrocyte-derived alarmin IL-33 mediates neuropathic pain. FASEB J 30:54–65. https://doi.org/10.1096/fj.14-267146
Zhao Y et al (2016) 6-C-(E-phenylethenyl)naringenin induces cell growth inhibition and cytoprotective autophagy in colon cancer cells. Eur J Cancer 68:38–50. https://doi.org/10.1016/j.ejca.2016.09.001
Zhou R, Tardivel A, Thorens B, Choi I, Tschopp J (2010) Thioredoxin-interacting protein links oxidative stress to inflammasome activation. Nat Immunol 11:136–140. https://doi.org/10.1038/ni.1831
Zhu L, Wang J, Wei T, Gao J, He H, Chang X, Yan T (2015) Effects of Naringenin on inflammation in complete freund’s adjuvant-induced arthritis by regulating Bax/Bcl-2 balance. Inflammation 38:245–251. https://doi.org/10.1007/s10753-014-0027-7
This work was supported by grants from Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq), Coordenacão de Aperfeiçoamento de Pessoal de Nível Superior (CAPES), Ministério da Ciência Tecnologia e Inovação (MCTI), Secretaria da Ciência, Tecnologia e Ensino Superior (SETI), Fundação Araucária, and Paraná State Government, Brazil. Sergio M. Borghi received a postdoctoral fellowship from CAPES and CNPq (152792/2016-3). The authors also thank the support of Central Multiusuário de Laboratórios de Pesquisa da Universidade Estadual de Londrina (CMLP-UEL).
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Bussmann, A.J.C., Borghi, S.M., Zaninelli, T.H. et al. The citrus flavanone naringenin attenuates zymosan-induced mouse joint inflammation: induction of Nrf2 expression in recruited CD45+ hematopoietic cells. Inflammopharmacol 27, 1229–1242 (2019). https://doi.org/10.1007/s10787-018-00561-6