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Research ArticleInflammation, Immunopharmacology, and Asthma

Anti-inflammatory Properties of Cannabidiol, a Nonpsychotropic Cannabinoid, in Experimental Allergic Contact Dermatitis

Stefania Petrosino, Roberta Verde, Massimo Vaia, Marco Allarà, Teresa Iuvone and Vincenzo Di Marzo
Journal of Pharmacology and Experimental Therapeutics June 2018, 365 (3) 652-663; DOI: https://doi.org/10.1124/jpet.117.244368

Article Figures & Data

Figures

  • Figure1
  • Fig. 1.
    Fig. 1.

    CBD reduces MCP-2 levels in poly-(I:C)–stimulated HaCaT cells. Enzyme-linked immunosorbent assay for MCP-2 release in the supernatants of poly-(I:C)–stimulated HaCaT cells (100 μg/ml) in the presence of vehicle or CBD (1, 5, 10, and 20 μM) for 6 (A), 12 (B), and 24 hours (C) at 37°C in 5% CO2. Data represent the mean ± S.E.M. of three independent experiments performed in triplicate. §§§P < 0.001 vs. vehicle; ***P < 0.001 vs. poly-(I:C). Assay range for MCP-2, 0.8–200 pg/ml.

  • Fig. 2.
    Fig. 2.

    CBC, CBG, THCV, and CBGV reduce MCP-2 levels in poly-(I:C)–stimulated HaCaT cells. Enzyme-linked immunosorbent assay for MCP-2 release in the supernatants of poly-(I:C)–stimulated HaCaT cells (100 μg/ml, 6 hours, 37°C) in the presence of vehicle or CBC (A), CBG (B), THCV (C), and CBGV (D) (all tested at 5, 10, and 20 μM). Data represent the mean ± S.E.M. of three independent experiments performed in triplicate. §§§P < 0.001 vs. vehicle; **P < 0.01; ***P < 0.001 vs. poly-(I:C). Assay range for MCP-2, 0.8–200 pg/ml.

  • Fig. 3.
    Fig. 3.

    CBD reduces IL-6, IL-8, and TNF-α levels in poly-(I:C)–stimulated HaCaT cells after 6 hours. Bio-Plex Pro assay for IL-6 (A), IL-8 (B), and TNF-α (C) release in the supernatants of poly-(I:C)–stimulated HaCaT cells (100 μg/ml, 6 hours, 37°C) in the presence of vehicle or CBD (1, 5, 10, and 20 μM). Data represent the mean ± S.E.M. of three independent experiments performed in triplicate. §§§ P < 0.001 vs. vehicle; *P < 0.05; ***P < 0.001 vs. poly-(I:C). Assay range for IL-6, 37.68 pg/ml; for IL-8, 42.15 pg/ml; and for TNF-α, 64.80 pg/ml.

  • Fig. 4.
    Fig. 4.

    CBD reduces IL-6 and TNF-α levels in poly-(I:C)–stimulated HaCaT cells after 12 and 24 hours. (A) Bio-Plex Pro assay for TNF-α release in the supernatants of poly-(I:C)–stimulated HaCaT cells (100 μg/ml, 12 hours, 37°C) in the presence of vehicle or CBD (20 μM). Bio-Plex Pro assay for IL-6 (B) and TNF-α (C) release in the supernatants of poly-(I:C)–stimulated HaCaT cells (100 μg/ml, 24 hours, 37°C) in the presence of vehicle or CBD (20 μM). Data represent the mean ± S.E.M. of three independent experiments performed in triplicate. §§§P < 0.001 vs. vehicle; ***P < 0.001 vs. poly-(I:C). Assay range for IL-6, 37.68 pg/ml; and for TNF-α, 64.80 pg/ml.

  • Fig. 5.
    Fig. 5.

    CBC, CBG, and THCV reduce IL-6 and IL-8 levels in poly-(I:C)–stimulated HaCaT cells after 6 hours. Bio-Plex Pro assay for IL-6 (A, C, and E) and IL-8 (B and D) release in the supernatants of poly-(I:C)–stimulated HaCaT cells (100 μg/ml, 6 hours, 37°C) in the presence of vehicle or CBC, CBG, and THCV (all tested at 5, 10, and 20 μM). Data represent the mean ± S.E.M. of three independent experiments performed in triplicate. §§§P < 0.001 vs. vehicle; *P < 0.05; **P < 0.01; ***P < 0.001 vs. poly-(I:C). Assay range for IL-6, 37.68 pg/ml; and for IL-8, 42.15 pg/ml.

  • Fig. 6.
    Fig. 6.

    CBD and other phytocannabinoids are not cytotoxic in HaCaT cells. (A) MTT assay in HaCaT cells treated with vehicle (white histogram) or CBD, CBC, CBG, THCV, and CBGV (all tested at 10 and 20 μM) for 6 hours at 37°C in 5% CO2. (B) MTT assay in HaCaT cells treated with vehicle or CBD (10 and 20 μM) for 12 and 24 hours at 37°C in 5% CO2. Data represent the mean ± S.E.M. of three independent experiments performed in triplicate.

  • Fig. 7.
    Fig. 7.

    CB2 and TRPV1 receptors mediate the action of CBD in poly-(I:C)–stimulated HaCaT cells after 6 hours. (A) Enzyme-linked immunosorbent assay for MCP-2 release in the supernatants of poly-(I:C)–stimulated HaCaT cells (100 μg/ml, 6 hours, 37°C) treated with AM251 (1, 2.5, and 5 μM) in the presence or absence of CBD (20 μM). (B) Enzyme-linked immunosorbent assay for MCP-2 release in the supernatants of poly-(I:C)–stimulated HaCaT cells (100 μg/ml, 6 hours, 37°C) treated with AM630 (0.01 and 0.1 μM) or I-RTX (0.01 and 0.1 μM) in the presence or absence of CBD (20 μM). Data represent the mean ± S.E.M. of three independent experiments performed in triplicate. §§§P < 0.001 vs. vehicle. ***P < 0.001; **P < 0.01 vs. poly-(I:C); °°P < 0.01; °°°P < 0.001 vs. poly-(I:C) + CBD 20 μM. Assay range for MCP-2, 0.8–200 pg/ml.

  • Fig. 8.
    Fig. 8.

    CBD elevates AEA levels in poly-(I:C)–stimulated HaCaT cells after 6 hours. Concentrations of AEA (A), 2-AG (B), PEA (C), and OEA (D) in poly-(I:C)–stimulated HaCaT cells (100 μg/ml, 6 hours, 37°C) in the presence of vehicle or CBD (20 μM). Data represent the mean ± S.E.M. of three independent experiments performed in triplicate. ***P < 0.001 vs. vehicle; °°°P < 0.001 vs. poly-(I:C) + vehicle.

  • Fig. 9.
    Fig. 9.

    AEA and URB597 reduce MCP-2, IL-6, and IL-8 levels in poly-(I:C)–stimulated HaCaT cells after 6 hours. Enzyme-linked immunosorbent assay for MCP-2 (A) and Bio-Plex Pro assay for IL-6 (B) and IL-8 (C) release in the supernatants of poly-(I:C)–stimulated HaCaT cells (100 μg/ml, 6 hours, 37°C) in the presence of vehicle or CBD (20 μM), AEA (10 μM), or URB597 (10 μM). Data represent the mean ± S.E.M. of three independent experiments performed in triplicate. §§§P < 0.001 vs. vehicle; *P < 0.05; **P < 0.01; ***P < 0.001 vs. poly-(I:C). Assay range for MCP-2, 0.8–200 pg/ml; for IL-6, 37.68 pg/ml; and for IL-8, 42.15 pg/ml.

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Research ArticleInflammation, Immunopharmacology, and Asthma

Cannabidiol and Allergic Contact Dermatitis

Stefania Petrosino, Roberta Verde, Massimo Vaia, Marco Allarà, Teresa Iuvone and Vincenzo Di Marzo
Journal of Pharmacology and Experimental Therapeutics June 1, 2018, 365 (3) 652-663; DOI: https://doi.org/10.1124/jpet.117.244368
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