Suppressive effects of antimycotics on thymic stromal lymphopoietin production in human keratinocytes

Abstract

Background: Thymic stromal lymphopoietin (TSLP) is produced by epidermal keratinocytes, and it induces Th2-mediated inflammation. TSLP expression is enhanced in lesions with atopic dermatitis, and is a therapeutic target. Antimycotic agents improve the symptoms of atopic dermatitis.

Objective: The objective of this study was to examine whether antimycotics suppress TSLP expression in human keratinocytes.

Methods: Normal human keratinocytes were incubated with polyinosinic-polycytidylic acid (poly I:C) plus IL-4 in the presence of antimycotics. TSLP expression was analyzed by ELISA and real time PCR. Luciferase assays were performed to analyze NF-κB activity. IκBα degradation was analyzed by Western blot analysis.

Results: Poly I:C plus IL-4 increased the secretion and mRNA levels of TSLP, which was suppressed by an NF-κB inhibitor, and also enhanced NF-κB transcriptional activities and induced the degradation of IκBα in keratinocytes. The antimycotics itraconazole, ketoconazole, luliconazole, terbinafine, butenafine, and amorolfine suppressed the secretion and mRNA expression of TSLP, NF-κB activity, and IκBα degradation induced by poly I:C plus IL-4. These suppressive effects were similarly manifested by 15-deoxy-Δ-(12,14)-PGJ2 (15d-PGJ2), a prostaglandin D2 metabolite. Antimycotics increased the release of 15d-PGJ2 from keratinocytes and decreased the release of thromboxane B2, a thromboxane A2 metabolite. Antimycotic-induced suppression of TSLP production and NF-κB activity was counteracted by an inhibitor of lipocalin type-prostaglandin D synthase.

Conclusions: Antimycotics itraconazole, ketoconazole, luliconazole, terbinafine, butenafine, and amorolfine may suppress poly I:C plus IL-4-induced production of TSLP by inhibiting NF-κB via increasing 15d-PGJ2 production in keratinocytes. These antimycotics may block the overexpression of TSLP in lesions with atopic dermatitis.

Keywords: 15-Deoxy-Δ-(12,14)-PGJ(2); 15-deoxy-Δ-(12,14)-prostaglandin J(2); 15d-PGJ(2); AD; ANOVA; Antimycotic; Atopic dermatitis; CRTH2; D prostanoid receptor; DMSO; DP; GAPDH; HPGDS; IKK; IκB; IκB kinase; KBM; Keratinocyte; LPGDS; NEMO; NF-κB; NF-κB essential modifier; PG; PPARγ; RIP1; TAK1; TGF-β activated kinase 1; TNF receptor; TNF receptor-associated death domain; TNF receptor-associated factor; TNFR; TRADD; TRAF; TRIF; TSLP; TXA(2); TXAS; Thymic stromal lymphopoietin; Toll/IL-1 receptor domain-containing adaptor inducing IFN-β; analysis of variance; atopic dermatitis; chemoattractant receptor-homologous molecule expressed on Th2 cells; dimethylsulfoxide; glyceraldehyde-3-phosphate dehydrogenase; hematopoietic prostaglandin D synthase; inhibitory κB; keratinocyte basal medium; lipocalin-type prostaglandin D synthase; peroxisome proliferator-activated receptor γ; poly I:C; polyinosinic–polycytidylic acid; prostaglandin; receptor interacting protein 1; thromboxane A(2); thromboxane A(2) synthase; thymic stromal lymphopoietin.