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

      Highlights

      • 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.

      Abstract

      Background

      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.

      Objective

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

      Methods

      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.

      Results

      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.

      Conclusion

      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.

      Abbreviations:

      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)

      Keywords

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