Oral ulcer is a common oral inflammatory lesion accompanied by severe pain but with few effective treatments. Cannabidiol (CBD) is recently emerging for its therapeutic potential in a range of diseases, including inflammatory conditions and cancers. Here we show that CBD oral spray on acid- or trauma-induced oral ulcers on mice tongue inhibits inflammation, relieves pain, and accelerates lesion closure. Notably, the enrichment of genes associated with the NOD, LRR, and NLRP3 pyrin domain–containing protein 3 (NLRP3) inflammasome pathway is downregulated after CBD treatment. The expression of cleaved-gasdermin D (GSDMD) and the percentage of pyroptotic cells are reduced as well. In addition, CBD decreases the expression of cytidine/uridine monophosphate kinase 2 (CMPK2), which subsequently inhibits the generation of oxidized mitochondria DNA and suppresses inflammasome activation. These immunomodulating effects of CBD are mostly blocked by peroxisome proliferator activated receptor γ (PPARγ) antagonist and partially antagonized by CB1 receptor antagonist. Our results demonstrate that CBD accelerates oral ulcer healing by inhibiting CMPK2-mediated NLRP3 inflammasome activation and pyroptosis, which are mediated mostly by PPARγ in the nucleus and partially by CB1 in the plasma membrane.
BidokiAZHarsiniSSadrMSoltaniSMohammadzadehMNajafiSRezaeiN.2016. NLRP3 gene polymorphisms in Iranian patients with recurrent aphthous stomatitis. J Oral Pathol Med. 45(2):136–140.
2.
BiroTTothBIHaskoGPausRPacherP.2009. The endocannabinoid system of the skin in health and disease: novel perspectives and therapeutic opportunities. Trends Pharmacol Sci. 30(8):411–420.
3.
CasaresLGarciaVGarrido-RodriguezMMillanEColladoJAGarcia-MartinAPenarandoJCalzadoMAde la VegaLMunozE.2020. Cannabidiol induces antioxidant pathways in keratinocytes by targeting BACH1. Redox Biol. 28:101321.
4.
DaganEGershoni-BaruchRKhatibIMoriABrikR.2010. MEFV, TNF1rA, CARD15 and NLRP3 mutation analysis in PFAPA. Rheumatol Int. 30(5):633–636.
5.
DanceA.2019. As CBD skyrockets in popularity, scientists scramble to understand how it’s metabolized. Nat Med. 25(10):1462–1464.
6.
DanilenkoDM.1999. Preclinical and early clinical development of keratinocyte growth factor, an epithelial-specific tissue growth factor. Toxicol Pathol. 27(1):64–71.
7.
De FilippisDEspositoGCirilloCCiprianoMDe WinterBYScuderiCSarnelliGCuomoRSteardoLDe ManJG, et al. 2011. Cannabidiol reduces intestinal inflammation through the control of neuroimmune axis. PLoS One. 6(12):e28159.
8.
Del RioCCantareroIPalomaresBGomez-CanasMFernandez-RuizJPavicicCGarcia-MartinALuz BellidoMOrtega-CastroRPerez-SanchezC, et al. 2018. VCE-004.3, a cannabidiol aminoquinone derivative, prevents bleomycin-induced skin fibrosis and inflammation through PPARγ- and CB2 receptor-dependent pathways. Br J Pharmacol. 175(19):3813–3831.
9.
DingJShaoF.2017. Snapshot: the noncanonical inflammasome. Cell. 168(3):544-544, e541.
10.
DuddingTHaworthSLindPASathirapongsasutiJFMe ResearchTTungJYMitchellRColodro-CondeLMedlandSEGordonS, et al. 2019. Genome wide analysis for mouth ulcers identifies associations at immune regulatory loci. Nat Commun. 10(1):1052.
11.
FriedmanDFrenchJAMaccarroneM.2019. Safety, efficacy, and mechanisms of action of cannabinoids in neurological disorders. Lancet Neurol. 18(5):504–512.
12.
HatemiGMahrAIshigatsuboYSongYWTakenoMKimDMelikogluMChengSMcCueSParisM, et al. 2019. Trial of apremilast for oral ulcers in Behcet’s syndrome. N Engl J Med. 381(20):1918–1928.
13.
Iglesias-BartolomeRPatelVCotrimALeelahavanichkulKMolinoloAAMitchellJBGutkindJS. 2012. mTOR inhibition prevents epithelial stem cell senescence and protects from radiation-induced mucositis. Cell Stem Cell. 11(3):401–414.
IrvingPMIqbalTNwokoloCSubramanianSBloomSPrasadNHartAMurrayCLindsayJOTaylorA, et al. 2018. A randomized, double-blind, placebo-controlled, parallel-group, pilot study of cannabidiol-rich botanical extract in the symptomatic treatment of ulcerative colitis. Inflamm Bowel Dis. 24(4):714–724.
16.
Jimenez-BlascoDBusquets-GarciaAHebert-ChatelainESerratRVicente-GutierrezCIoannidouCGomez-SotresPLopez-FabuelIResch-BeusherMReselE, et al. 2020. Glucose metabolism links astroglial mitochondria to cannabinoid effects. Nature. 583(7817):603–608.
17.
KarsakMGaffalEDateRWang-EckhardtLRehneltJPetrosinoSStarowiczKSteuderRSchlickerECravattB, et al. 2007. Attenuation of allergic contact dermatitis through the endocannabinoid system. Science. 316(5830):1494–1497.
18.
KawaiTAkiraS.2010. The role of pattern-recognition receptors in innate immunity: update on Toll-like receptors. Nat Immunol. 11(5):373–384.
19.
KleinMde QuadrosDeBortolliJGuimaraesFSSalumFGCherubiniKde FigueiredoMAZ. 2018. Effects of cannabidiol, a Cannabis sativa constituent, on oral wound healing process in rats: clinical and histological evaluation. Phytother Res. 32(11):2275–2281.
20.
KozelaEPietrMJuknatARimmermanNLevyRVogelZ.2010. Cannabinoids delta(9)-tetrahydrocannabinol and cannabidiol differentially inhibit the lipopolysaccharide-activated NF-kappaB and interferon-beta/STAT proinflammatory pathways in BV-2 microglial cells. J Biol Chem. 285(3):1616–1626.
21.
LiouGIAuchampachJAHillardCJZhuGYousufzaiBMianSKhanSKhalifaY.2008. Mediation of cannabidiol anti-inflammation in the retina by equilibrative nucleoside transporter and A2A adenosine receptor. Invest Ophthalmol Vis Sci. 49(12):5526–5531.
22.
LiuCMaHSlittALSeeramNP. 2020. Inhibitory effect of cannabidiol on the activation of NLRP3 inflammasome is associated with its modulation of the P2X7 receptor in human monocytes. J Nat Prod. 83(6):2025–2029.
23.
LiuXZhangZRuanJPanYMagupalliVGWuHLiebermanJ.2016. Inflammasome-activated gasdermin D causes pyroptosis by forming membrane pores. Nature. 535(7610):153–158.
24.
Munoz-CorcueraMEsparza-GomezGGonzalez-MolesMABascones-MartinezA.2009. Oral ulcers: clinical aspects. A tool for dermatologists: part I. Acute ulcers. Clin Exp Dermatol. 34(3):289–294.
25.
NaftaliTLevLBYablecovitchDHalfEKonikoffFM. 2011. Treatment of Crohn’s disease with cannabis: an observational study. Isr Med Assoc J. 13(8):455–458.
26.
OlahATothBIBorbiroISugawaraKSzollosiAGCzifraGPalBAmbrusLKloepperJCameraE, et al. 2014. Cannabidiol exerts sebostatic and antiinflammatory effects on human sebocytes. J Clin Invest. 124(9):3713–3724.
27.
OrtizFAcuna-CastroviejoDDoerrierCDayoubJCLopezLCVenegasCGarciaJALopezAVoltHLuna-SanchezM, et al. 2015. Melatonin blunts the mitochondrial/NLRP3 connection and protects against radiation-induced oral mucositis. J Pineal Res. 58(1):34–49.
28.
PisantiSMalfitanoAMCiagliaELambertiARanieriRCuomoGAbateMFaggianaGProtoMCFioreD, et al. 2017. Cannabidiol: state of the art and new challenges for therapeutic applications. Pharmacol Ther. 175:133–150.
29.
RajanJVRodriguezDMiaoEAAderemA.2011. The NLRP3 inflammasome detects encephalomyocarditis virus and vesicular stomatitis virus infection. J Virol. 85(9):4167–4172.
30.
RibeiroAAlmeidaVICostola-de-SouzaCFerraz-de-PaulaVPinheiroMLVitorettiLBGimenes-JuniorJAAkamineATCrippaJATavares-de-LimaW, et al. 2015. Cannabidiol improves lung function and inflammation in mice submitted to LPS-induced acute lung injury. Immunopharmacol Immunotoxicol. 37(1):35–41.
31.
SacerdotePMartucciCVaccaniABariselliFPaneraiAEColomboAParolaroDMassiP.2005. The nonpsychoactive component of marijuana cannabidiol modulates chemotaxis and IL-10 and IL-12 production of murine macrophages both in vivo and in vitro. J Neuroimmunol. 159(1–2):97–105.
32.
SharmaRTobinPClarkeSJ. 2005. Management of chemotherapy-induced nausea, vomiting, oral mucositis, and diarrhoea. Lancet Oncol. 6(2):93–102.
33.
SlezakovaSBorilova LinhartovaPMasopustovaLBartovaJPetanovaJKuklinekPFassmannADusekLIzakovicova HollaL.2018. Association of the NOD-like receptor 3 (NLRP3) gene variability with recurrent aphthous stomatitis in the Czech population. J Oral Pathol Med. 47(4):434–439.
34.
SohnE.2015. Israel: research without prejudice. Nature. 525(7570):S12–S13.
35.
SonegoABPradoDSValeGTSepulveda-DiazJECunhaTMTirapelliCRDel BelEARaisman-VozariRGuimaraesFS. 2018. Cannabidiol prevents haloperidol-induced vacuos chewing movements and inflammatory changes in mice via PPARγ receptors. Brain Behav Immun. 74:241–251.
36.
SonisST.2004. The pathobiology of mucositis. Nat Rev Cancer. 4(4):277–284.
37.
SonisST.2010. Efficacy of palifermin (keratinocyte growth factor-1) in the amelioration of oral mucositis. Core Evid. 4:199–205.
38.
SwansonKVDengMTingJP. 2019. The NLRP3 inflammasome: molecular activation and regulation to therapeutics. Nat Rev Immunol. 19(8):477–489.
39.
ZhongZLiangSSanchez-LopezEHeFShalapourSLinXJWongJDingSSekiESchnablB, et al. 2018. New mitochondrial DNA synthesis enables NLRP3 inflammasome activation. Nature. 560(7717):198–203.
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