피부 혈관이 피부 색소변화에 끼치는 영향
The effect of cutaneous vasculature on skin pigmentation
- 주제(키워드) Co-culture , Cutaneous vasculature , Endothelial cells , Hyperpigmentary disorder , Melanocytes , Melasma , Skin pigmentation
- 발행기관 아주대학교
- 지도교수 강희영
- 발행년도 2015
- 학위수여년월 2015. 8
- 학위명 박사
- 학과 및 전공 일반대학원 의학과
- 실제URI http://www.dcollection.net/handler/ajou/000000020277
- 본문언어 영어
- 저작권 아주대학교 논문은 저작권에 의해 보호받습니다.
초록/요약
Previously, it was demonstrated that a hyperpigmented melasma lesion is characterized by increased vasculature, which may play a role in the development of hyperpigmentation. Herein, I report the in vitro evidence of the vascular modulation of skin pigmentation. In the presence of endothelial cells, the pigmentation of melanocytes and of ex vivo cultured human skin was reduced. Contrary to non-stimulated cells, UV-irradiated endothelial cells showed stimulatory action on skin pigmentation. However, other stimulation, such as VEGF treatment and hypoxic condition, did not shown any significant effect. Gene expression profiling was undertaken to identify melanogenic factors from UV-irradiated endothelial cells. Endothelin-1 (ET-1) was found to be up-regulated, and the increased pigmentation elicited by UV-irradiated endothelial cell-derived conditioned medium was prevented by inhibiting the ET-1 receptor. Our study suggests that endothelial cells have a negative effect on pigmentation. However, their effect differs after they are irradiated with UV, as they then positively influence the induction of melanogenesis. This may play a role in the development of hyperpigmentary disorders.
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ABSTARCT i
TABLE OF CONTENTS ii
LIST OF FIGURES iv
LIST OF TABLES vii
I. INTRODUCTION 1
II. MATERIALS AND METHODS 3
A. Cell cultures and treatment 3
B. Co-culture using transwell system 4
C. Preparation of the bEnd.3/HUVECs conditioned medium 4
D. Ultraviolet (UV) irradiation of endothelial cells 5
E. Endothelial tube formation assay model using GeltrexTM Matrix 5
F. Hypoxia treatments 5
G. Melanin content and tyrosinase activity 5
H. Western blot analysis 6
I. Real-time PCR and RT-PCR 6
J. RNA sequence analysis 7
K. Ex vivo skin organ culture and pigmentation assay of cultured skin 7
L. Statistical analysis 8
III. RESULTS 9
A. Preliminary study 9
1. Effect of HUVECs on the proliferation of fibroblasts 9
B. The presence of endothelial cells has a negative effect on pigmentation 10
1. The effect of murine endothelial cells on pigmentation 10
2. The verification of the effect of endothelial cells with human cells 12
C. Additional modification of endothelial cells 15
1. UV irradiation: Contrary to non-stimulated cells, UV-irradiated endothelial cells induce pigmentation 15
2. VEGF treatment 19
3. Endothelial tube formation assay using GeltrexTM Matrix 20
4. Endothelial cell-conditioned media with hypoxic condition 22
5. Triple co-culture study with NHMs, HUVECs, and fibroblasts 23
D. Identification of possible melanogenic factors secreted by endothelial cells 24
1. Identification of endothelin-1 as one of melanogenic factors secreted by UV-irradiated endothelial cells 24
2. Identification of candidate cytokines derived from HUVECs using angiogenesis-related cytokine antibody array 28
IV. DISCUSSION 31
V. CONCLUSION 36
REFERENCES 37
국문요약 44
