Cannabidiol mediates epidermal terminal differentiation and redox homeostasis through aryl hydrocarbon receptor (AhR)-dependent signaling


      • CBD regulates AhR in keratinocytes and epidermal equivalents.
      • CBD induces epidermal differentiation by modulating the AhR-OVOL1 axis.
      • AhR orchestrates ROS scavenging through NRF2 regulation.



      Cannabidiol, a non-psychoactive phytocannabinoid, has antioxidant and anti-inflammatory activity in keratinocytes. However, the signaling pathway through which cannabidiol exerts its effect on keratinocytes or whether it can modulate keratinocyte differentiation has not been fully elucidated yet.


      We investigated whether cannabidiol modulates epidermal differentiation and scavenges reactive oxygen species through the aryl hydrocarbon receptor (AhR) in keratinocytes and epidermal equivalents.


      We investigated the cannabidiol-induced activation of AhR using AhR luciferase reporter assay, qRT-PCR, western blot, and immunofluorescence assays. We also analyzed whether keratinocyte differentiation and antioxidant activity are regulated by cannabidiol-induced AhR activation.


      In both keratinocytes and epidermal equivalents, cannabidiol increased both the mRNA and protein expression of filaggrin, involucrin, NRF2, and NQO1 and the mRNA expression of the AhR target genes, including CYP1A1 and aryl hydrocarbon receptor repressor. Additionally, cannabidiol showed antioxidant activity that was attenuated by AhR knockdown or co-administration with an AhR antagonist. Moreover, cannabidiol increased the ratio of OVOL1/OVOL2 mRNA expression, which is a downstream regulator of AhR that mediates epidermal differentiation. In addition to increased expression of barrier-related proteins, cannabidiol-treated epidermal equivalent showed a more prominent granular layer than the control epidermis. The increased granular layer by cannabidiol was suppressed by the AhR antagonist.


      Cannabidiol can be a modulator of the AhR-OVOL1-filaggrin axis and AhR-NRF2-NQO1 signaling, thus indicating a potential use of cannabidiol in skin barrier enhancement and reducing oxidative stress.


      CBD (Cannabidiol), EB (epidermolysis bullosa), AhR (Aryl hydrocarbon receptor), TCDD (2,3,7,8-tetracholrodibenzo-p-dioxin), FLG (filaggrin), OVOL1 (OVO like 1), OVOL2 (OVO like 2), FICZ (6-formylindolo-(3,2-b)-carbazole), NRF2 (nuclear factor erythroid-2-related factor 2), NQO1 (NAD(P)H quinone oxidoreductase 1), CYP1A1 (cytochrome P450 family-1 subfamily-A polypeptide-1), NHEKs (Normal human epidermal keratinocytes), DMSO (Dimethyl sulfoxide), CB (Cannabinoid receptor), TRPV1 (Transient receptor potential cation channel subfamily V member 1), ARNT (AhR nuclear translocator), FLG (Filaggrin), LOR (Lorricrin), IVL (involucrin), BaP (Benzo(a)pyrene), ROS (Reactive oxygen species), t-BHP (tert-Butyl hydroperoxide), HO-1 (Heme oxygenase-1)


      To read this article in full you will need to make a payment

      Purchase one-time access:

      Academic & Personal: 24 hour online accessCorporate R&D Professionals: 24 hour online access
      One-time access price info
      • For academic or personal research use, select 'Academic and Personal'
      • For corporate R&D use, select 'Corporate R&D Professionals'


      Subscribe to Journal of Dermatological Science
      Already a print subscriber? Claim online access
      Already an online subscriber? Sign in
      Institutional Access: Sign in to ScienceDirect


        • Karl T.
        • Garner B.
        • Cheng D.
        The therapeutic potential of the phytocannabinoid cannabidiol for Alzheimer's disease.
        Behav. Pharm. 2017; 28: 142-160
        • Watt G.
        • Karl T.
        In vivo evidence for therapeutic properties of cannabidiol (CBD) for Alzheimer's disease.
        Front Pharm. 2017; 8: 20
        • Franco V.
        • Bialer M.
        • Perucca E.
        Cannabidiol in the treatment of epilepsy: Current evidence and perspectives for further research.
        Neuropharmacology. 2021; 185108442
        • Mao K.
        • et al.
        High dosage of cannabidiol (CBD) alleviates pentylenetetrazole-induced epilepsy in rats by exerting an anticonvulsive effect.
        Int J. Clin. Exp. Med. 2015; 8: 8820-8827
        • Giacoppo S.
        • et al.
        Target regulation of PI3K/Akt/mTOR pathway by cannabidiol in treatment of experimental multiple sclerosis.
        Fitoterapia. 2017; 116: 77-84
        • Flachenecker P.
        • Henze T.
        • Zettl U.K.
        Nabiximols (THC/CBD oromucosal spray, Sativex®) in clinical practice--results of a multicenter, non-interventional study (MOVE 2) in patients with multiple sclerosis spasticity.
        Eur. Neurol. 2014; 71: 271-279
        • Gugliandolo A.
        • et al.
        Cannabidiol exerts protective effects in an in vitro model of Parkinson's disease activating AKT/mTOR pathway.
        Fitoterapia. 2020; 143104553
        • Zuardi A.W.
        • et al.
        Cannabidiol for the treatment of psychosis in Parkinson's disease.
        J. Psychopharmacol. 2009; 23: 979-983
        • Xu D.H.
        • et al.
        The effectiveness of topical cannabidiol oil in symptomatic relief of peripheral neuropathy of the lower extremities.
        Curr. Pharm. Biotechnol. 2020; 21: 390-402
        • Hoggart B.
        • et al.
        A multicentre, open-label, follow-on study to assess the long-term maintenance of effect, tolerance and safety of THC/CBD oromucosal spray in the management of neuropathic pain.
        J. Neurol. 2015; 262: 27-40
        • Maghfour J.
        • et al.
        Assessing the effects of topical cannabidiol in patients with atopic dermatitis.
        Dermatol. Online J. 2021; 27
        • Gao Y.
        • et al.
        Novel cannabidiol aspartame combination treatment (JW-100) significantly reduces ISGA score in atopic dermatitis: Results from a randomized double-blinded placebo-controlled interventional study.
        J. Cosmet. Dermatol. 2022; 21: 1647-1650
        • Vincenzi C.
        • Tosti A.
        Efficacy and tolerability of a shampoo containing broad-spectrum cannabidiol in the treatment of scalp inflammation in patients with mild to moderate scalp psoriasis or seborrheic dermatitis.
        Ski. Appendage Disord. 2020; 6: 355-361
        • Jarocka-Karpowicz I.
        • et al.
        Cannabidiol effects on phospholipid metabolism in keratinocytes from patients with psoriasis vulgaris.
        Biomolecules. 2020; 10
        • Chelliah M.P.
        • et al.
        Self-initiated use of topical cannabidiol oil for epidermolysis bullosa.
        Pedia Dermatol. 2018; 35: e224-e227
        • Shao K.
        • Stewart C.
        • Grant-Kels J.M.
        Cannabis and the skin.
        Clin. Dermatol. 2021; 39: 784-795
        • Atalay S.
        • Jarocka-Karpowicz I.
        • Skrzydlewska E.
        Antioxidative and anti-inflammatory properties of cannabidiol.
        Antioxid. (Basel). 2019; 9
        • Jastrząb A.
        • et al.
        Antioxidant and anti-inflammatory effect of cannabidiol contributes to the decreased lipid peroxidation of keratinocytes of rat skin exposed to UV radiation.
        Oxid. Med. Cell. Longev. 2021; 2021: 6647222
        • Fumagalli M.
        • et al.
        Cannabidiol has a central role in the NF-κB inhibition of a Cannabis sativa L. extract in human keratinocytes. Zeitschrift für Phytotherapie.
        . 2022; 43: P36
        • Casares L.
        • et al.
        Cannabidiol induces antioxidant pathways in keratinocytes by targeting BACH1.
        Redox Biol. 2020; 28101321
        • Poland A.
        • Knutson J.C.
        2,3,7,8-tetrachlorodibenzo-p-dioxin and related halogenated aromatic hydrocarbons: examination of the mechanism of toxicity.
        Annu Rev. Pharm. Toxicol. 1982; 22: 517-554
        • Gutiérrez-Vázquez C.
        • Quintana F.J.
        Regulation of the immune response by the aryl hydrocarbon receptor.
        Immunity. 2018; 48: 19-33
        • Kyoreva M.
        • et al.
        CYP1A1 enzymatic activity influences skin inflammation via regulation of the AHR pathway.
        J. Invest Dermatol. 2021; 141 (e3): 1553-1563
        • Haas K.
        • et al.
        Aryl hydrocarbon receptor in keratinocytes is essential for murine skin barrier integrity.
        J. Invest Dermatol. 2016; 136: 2260-2269
        • Tsuji G.
        • et al.
        The role of the OVOL1-OVOL2 axis in normal and diseased human skin.
        J. Dermatol. Sci. 2018; 90: 227-231
        • Takei K.
        • et al.
        Antioxidant soybean tar Glyteer rescues T-helper-mediated downregulation of filaggrin expression via aryl hydrocarbon receptor.
        J. Dermatol. 2015; 42: 171-180
        • Smith S.H.
        • et al.
        Tapinarof Is a Natural AhR Agonist that Resolves Skin Inflammation in Mice and Humans.
        J. Invest Dermatol. 2017; 137: 2110-2119
        • Kiyomatsu-Oda M.
        • et al.
        Protective role of 6-formylindolo[3,2-b]carbazole (FICZ), an endogenous ligand for arylhydrocarbon receptor, in chronic mite-induced dermatitis.
        J. Dermatol. Sci. 2018; 90: 284-294
        • Yamaori S.
        • et al.
        Cannabidiol induces expression of human cytochrome P450 1A1 that is possibly mediated through aryl hydrocarbon receptor signaling in HepG2 cells.
        Life Sci. 2015; 136: 87-93
        • Kupczyk P.
        • Reich A.
        • Szepietowski J.C.
        Cannabinoid system in the skin - a possible target for future therapies in dermatology.
        Exp. Dermatol. 2009; 18: 669-679
        • Hashimoto-Hachiya A.
        • et al.
        Upregulation of FLG, LOR, and IVL expression by rhodiola crenulata root extract via aryl hydrocarbon receptor: differential involvement of OVOL1.
        Int J. Mol. Sci. 2018; 19
        • Dietrich C.
        Antioxidant functions of the aryl hydrocarbon receptor.
        Stem Cells Int. 2016; 2016: 7943495
        • Jastrząb A.
        • Gęgotek A.
        • Skrzydlewska E.
        Cannabidiol Regulates the Expression of Keratinocyte Proteins Involved in the Inflammation Process through Transcriptional Regulation.
        Cells. 2019; 8
        • Borges R.S.
        • et al.
        Understanding the molecular aspects of tetrahydrocannabinol and cannabidiol as antioxidants.
        Molecules. 2013; 18: 12663-12674
        • Singer E.
        • et al.
        Reactive oxygen species-mediated therapeutic response and resistance in glioblastoma.
        Cell Death Dis. 2015; 6e1601
        • Tóth B.I.
        • et al.
        Endocannabinoids modulate human epidermal keratinocyte proliferation and survival via the sequential engagement of cannabinoid receptor-1 and transient receptor potential vanilloid-1.
        J. Invest Dermatol. 2011; 131: 1095-1104
        • Maccarrone M.
        • et al.
        The endocannabinoid system in human keratinocytes. Evidence that anandamide inhibits epidermal differentiation through CB1 receptor-dependent inhibition of protein kinase C, activation protein-1, and transglutaminase.
        J. Biol. Chem. 2003; 278: 33896-33903
        • Paradisi A.
        • et al.
        Anandamide regulates keratinocyte differentiation by inducing DNA methylation in a CB1 receptor-dependent manner.
        J. Biol. Chem. 2008; 283: 6005-6012
        • Pertwee R.G.
        The diverse CB1 and CB2 receptor pharmacology of three plant cannabinoids: delta9-tetrahydrocannabinol, cannabidiol and delta9-tetrahydrocannabivarin.
        Br. J. Pharm. 2008; 153: 199-215
        • Thomas A.
        • et al.
        6"-Azidohex-2"-yne-cannabidiol: a potential neutral, competitive cannabinoid CB1 receptor antagonist.
        Eur. J. Pharm. 2004; 487: 213-221
        • Wilkinson J.D.
        • Williamson E.M.
        Cannabinoids inhibit human keratinocyte proliferation through a non-CB1/CB2 mechanism and have a potential therapeutic value in the treatment of psoriasis.
        J. Dermatol. Sci. 2007; 45: 87-92
        • Wells J.
        • et al.
        Ovol2 suppresses cell cycling and terminal differentiation of keratinocytes by directly repressing c-Myc and Notch1.
        J. Biol. Chem. 2009; 284: 29125-29135
        • Tsuji G.
        • et al.
        Aryl hydrocarbon receptor activation restores filaggrin expression via OVOL1 in atopic dermatitis.
        Cell Death Dis. 2017; 8e2931
        • Schräder N.H.B.
        • et al.
        Combined tetrahydrocannabinol and cannabidiol to treat pain in epidermolysis bullosa: a report of three cases.
        Br. J. Dermatol. 2019; 180: 922-924
        • Jeong S.
        • et al.
        Cannabidiol overcomes oxaliplatin resistance by enhancing NOS3- and SOD2-induced autophagy in human colorectal cancer cells.
        Cancers (Basel). 2019; 11
        • Jeong S.
        • et al.
        Cannabidiol-induced apoptosis is mediated by activation of Noxa in human colorectal cancer cells.
        Cancer Lett. 2019; 447: 12-23
        • Olivas-Aguirre M.
        • et al.
        Cannabidiol directly targets mitochondria and disturbs calcium homeostasis in acute lymphoblastic leukemia.
        Cell Death Dis. 2019; 10: 779
        • Zhang X.
        • et al.
        Cannabidiol induces cell cycle arrest and cell apoptosis in human gastric cancer SGC-7901 cells.
        Biomolecules. 2019; 9
        • Souza J.D.S.
        • et al.
        Case report: cannabidiol-induced skin rash: a case series and key recommendations.
        Front Pharm. 2022; 13881617