- 作者：Alison R. Forrester ; Martina S. Elias ; Emma L. Woodward ; Mark Graham ; Faith M. Williams ; Nick J. Reynolds
- 关键词：AhR ; aryl hydrocarbon receptor ; AHRR ; aryl hydrocarbon repressor protein ; 伪-NF ; 伪-naphthoflavone ; 尾-NF ; 尾-naphthoflavone ; CYP1A1 ; cytochrome P450 1A1 ; ITE ; 2-(1&prime ; H-indole-3&prime ; -carbonyl)-thiazole-4-carboxylic acid methyl ester ; TCDD ; 2 ; 3 ; 7 ; 8-tetrachlorodibenzo-p-dioxin ; TGM-1 ; transglutaminase-1 ; XRE ; xenobiotic response element
- 刊名：Journal of Dermatological Science
- 出版年：January, 2014
- 年：2014
- 卷：73
- 期：1
- 页码：10-22
- 全文大小：3111 K
Background
2,3,7,8-Tetrachlorodibenzo-p-dioxin (TCDD) is a potent activator of the aryl hydrocarbon receptor (AhR) and causes chloracne in humans. The pathogenesis and role of AhR in chloracne remains incompletely understood.Objective
To elucidate the mechanisms contributing to the development of the chloracne-like phenotype in a human epidermal equivalent model and identify potential biomarkers.
Methods
Using primary normal human epidermal keratinocytes (NHEK), we studied AhR activation by XRE-luciferase, AhR degradation and CYP1A1 induction. We treated epidermal equivalents with high affinity TCDD or two non-chloracnegens: 尾-naphthoflavone (尾-NF) and 2-(1鈥睭-indole-3鈥?carbonyl)-thiazole-4-carboxylic acid methyl ester (ITE). Using Western blotting and immunochemistry for filaggrin (FLG), involucrin (INV) and transglutaminase-1 (TGM-1), we compared the effects of the ligands on keratinocyte differentiation and development of the chloracne-like phenotype by H&E.
Results
In NHEKs, activation of an XRE-luciferase and CYP1A1 protein induction correlated with ligand binding affinity: TCDD > 尾-NF > ITE. AhR degradation was induced by all ligands. In epidermal equivalents, TCDD induced a chloracne-like phenotype, whereas 尾-NF or ITE did not. All three ligands induced involucrin and TGM-1 protein expression in epidermal equivalents whereas FLG protein expression decreased following treatment with TCDD and 尾-NF. Inhibition of AhR by 伪-NF blocked TCDD-induced AhR activation in NHEKs and blocked phenotypic changes in epidermal equivalents; however, AhR knock down did not reproduce the phenotype.
Conclusion
Ligand-induced CYP1A1 and AhR degradation did not correlate with their chloracnegenic potential, indicating that neither CYP1A1 nor AhR are suitable biomarkers. Mechanistic studies showed that the TCDD-induced chloracne-like phenotype depends on AhR activation whereas AhR knock down did not appear sufficient to induce the phenotype.