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Cis-Urocanic Acid Stimulates Primary Human Keratinocytes Independently of Serotonin or Platelet-Activating Factor Receptors  Kazuyo Kaneko, Jeffrey B.

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Presentation on theme: "Cis-Urocanic Acid Stimulates Primary Human Keratinocytes Independently of Serotonin or Platelet-Activating Factor Receptors  Kazuyo Kaneko, Jeffrey B."— Presentation transcript:

1 cis-Urocanic Acid Stimulates Primary Human Keratinocytes Independently of Serotonin or Platelet-Activating Factor Receptors  Kazuyo Kaneko, Jeffrey B. Travers, Mary S. Matsui, Antony R. Young, Mary Norval, Susan L. Walker  Journal of Investigative Dermatology  Volume 129, Issue 11, Pages (November 2009) DOI: /jid Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

2 Figure 1 mRNA expression of 5HT2A and PAF receptors in primary human keratinocytes and fibroblasts. 5HT2A (open column) and PAF (shaded column) receptor mRNA expressions were quantified using real-time RT-PCR (TaqMan). The results are expressed as mean±SD of three donors. ND, not detected. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

3 Figure 2 5HT receptor antagonist has no inhibitory effect on cis-urocanic acid-induced mediator production in primary human keratinocytes. Keratinocytes were pretreated with 10μM Metergoline (shaded column) or ethanol vehicle (open column) for 1hour before treatment with PBS, cis-UCA (700μM), or 5HT (1mM) for 24hours. Cell culture supernatants were collected and analyzed for (a) PGE2, (b) TNF-α, and (c) IL-6 by ELISA. The results are expressed as mean±SD (n=3) of one representative experiment of three. *P<0.05, **P<0.01, and ***P<0.001 compared with PBS control. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

4 Figure 3 cis-Urocanic acid is unlikely to stimulate the PAF receptor and has minimal effects on PAF-receptor-mediated PGE2 production in the KB PAF receptor model system. (a) Calcium mobilization responses of PAF-receptor-positive KBP and PAF-receptor-negative KBM cells in response to cis-UCA and trans-UCA (140μM) and PAF (100nM). Fluorescence intensity of Ca2+-sensitive dye Fura-2 was measured over time with a spectrophotofluorometer. The data represent one of three independent experiments. (b) Displacement of radioligand binding (10nM [3H]WEB2086) from the PAF receptor by cis-UCA (50–500μM) and trans-UCA (500μM), and by PAF (1μM) in KBP cells. (c) PGE2 release from KBP (shaded column) and KBM (open column) cells treated with PBS, cis-UCA (70 and 700μM) or trans-UCA (700μM), or with PAF (100nM) for 24hours. PGE2 concentrations in cell culture supernatants were assessed by ELISA. Results are expressed as mean±SD (n=3) of one representative experiment of three. ***P<0.001 compared with PBS control. NS, not significant. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

5 Figure 4 PAF receptor antagonist does not inhibit PGE2 increase by cis-urocanic acid in primary human keratinocytes. Keratinocytes were pretreated with 10μM ABT-491 (shaded column) or with water vehicle (open column) for 1hour before treatment with PBS, cis-UCA (70 or 700μM), or PAF (1μM) for 24hours. Cell culture supernatants were collected and analyzed for PGE2 by ELISA. Results are expressed as mean±SD (n=3) of one representative experiment of three. *P<0.05, **P<0.01, and ***P<0.001 compared with PBS control. Journal of Investigative Dermatology  , DOI: ( /jid ) Copyright © 2009 The Society for Investigative Dermatology, Inc Terms and Conditions

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