Regulatory mechanism of TSLP-induced filaggrin in human keratinocytes
피부각질세포에서 thymic stromal lymphopoietin (TSLP) 에 의한 filaggrin 발현 조절 기전에 대한 연구
- 주제(키워드) atopic dermatitis (AD) , ERK , filaggrin , keratinocytes , skin barrier , STAT3 , thymic stromal lymphopoietin (TSLP)
- 발행기관 고려대학교 대학원
- 지도교수 계영철
- 발행년도 2014
- 학위수여년월 2014. 2
- 학위구분 박사
- 학과 일반대학원 의학과
- 세부전공 피부과학전공
- 원문페이지 37 p
- 실제URI http://www.dcollection.net/handler/korea/000000048955
- 본문언어 영어
- 제출원본 000045795077
초록/요약
Recent studies have reported that inflammatory cytokines downregulate filaggrin expression. Meanwhile, thymic stromal lymphopoietin (TSLP), which skews the immune reaction in Th2 direction, is a critical cytokine in development of allergic diseases including atopic dermatitis. Atopic dermatitis (AD) is a chronic inflammatory skin disease that is characterized by a defective skin barrier and Th2 immune response. Most patients with AD show reduced expression of filaggrin, which plays an important role to maintain intact skin barrier function. The aim of this study was to investigate whether TSLP can modulate filaggrin expression in keratinocytes. To my knowledge, the presence of TSLP receptors in keratinocytes was first demonstrated in this study. It was also found that filaggrin expression was significantly reduced by TSLP in human keratinocytes and this process was mediated by STAT3- and/or ERK1/2-dependent signaling pathways in this study. Further, this study demonstrated the effect of TSLP on filaggrin expression in in vivo mouse experiment. The skin samples from TSLP-treated mice showed decreased filaggrin expression, thickened epidermis, and increased transepidermal water loss, indicative of a disrupted barrier function. These results suggest that TSLP and the related downstream molecules might be a target to ameliorate the disrupted barrier in patients with AD.
more목차
1. Abstract 1
2. Introduction 3
3. Materials and methods 6
1) Culture and stimulation of primary human keratinocytes 6
2) Preparation of siRNAs and transfection 6
3) RNA extraction and RT-PCR 7
4) Western blot 8
5) Immunofluorescence (IF) and confocal microscopy 9
6) Mice and collection of skin biopsies 9
7) Immunohistochemistry 10
8) Measurement of transepidermal water loss (TEWL) 10
9) Statistical Analysis 11
4. Results 12
1) Keratinocytes express TSLP receptor complex 12
2) TSLP inhibits filaggrin expression in keratinocytes in a dose-dependent manner 12
3) Comparison of inhibitory effects of TSLP and IL-4, IL-17 and IL-22 on the expression of filaggrin 13
4) STAT3 and ERK1/2 signaling pathways are activated in TSLP-treated keratinocytes 14
5) Inhibitors against STAT3 and/or ERK1/2 suppress TSLP-induced filaggrin downregulation in keratinocytes 14
6) siRNA-mediated STAT3- and/or ERK1/2-silencing abolishes the inhibitory action of TSLP on filaggrin expression 15
7) Epidermal filaggrin expression is suppressed by TSLP in vivo 15
5. Discussion 17
6. References 21
