彈力素 遺傳子 發現에 interleukin-1ß와 interleukin-10이 미치는 影響

다국어 초록 (Multilingual Abstract)

The elastic fibers are a major fibrillar component of the extracellular matrix of several organs. and their presence provides elastic properties to these tissues. Recent molecular biological studies also have revealed that accumulation of elastotic material in the skin reflects enhanced synthesis of not only elastin but also a fibrillin, an integral component of the elastic fibers, and steady-state levels of the elastin mRNA increased in photo damaged skin. Furthermore a variety of cytokines. Growth factors, and hormones have been shown to modulate elastin gene expression including transforming growth factor-ß, tumor necrosis factor^(-a), insulin-like growth factor-I, glucocorticoids, cyclic AMP, and vitamin D3. Recently, interleukin-1ß(ⅠL-1ß) and interleukin-10 (ⅠL-1 0)have been shown to up-regulatory function in elastin gene expression. In this study, the effects of IL-1ß and IL-10 were examined on elastin gene expression by Northern blot analysis. chloramphenicol acetyltransferase(CAT) assay, and laser confocal microscopy in cultured human fibroblasts. In Northern blot analysis. IL-1β showed an up-regulatory effect on elastin mRNA steady-state levels 2.09 and 2.54-fold compared with control cultured and incubated without IL-1ß and IL-10 showed an up-regulatory effect on elastin mRNA steady-state levels 1.09, 3.18 and 3.45-fold compared with control cultured and incubated without IL-10. In CAT assay, the percentage of acetylation were 8.6% in untreated control and 13.2% in 1U/ml of IL-1,8 and18.9% in 10U/ml of IL-1ß and 19.2% in 1ng/ml of IL-10 and 19% in 10ng/ml of IL10 in cultured fibroblasts. The promoter activities were increased 1.5 and 2. 2-fold in IL- 1β (1U/ml. 10U/ml) and 2.23 and 2.21-fold in IL-10 (lng/ml. 10ng/ml) treated fibroblasts compared to control groups. It means that IL-1, 8 and IL-10 enhanced elastin gene promoter activity. In laser confocal microscopy, the results indicated significant increase in the elastin epitopes associated with the cells cultured with IL-1ß and IL-10.
These results show that IL-1ß and IL-10 increase elastin gene expression on transcriptional and protein levels by cultured fibroblasts. It suggests that they may effect on elastin synthesis by a stage up-regulator in human skin.

목차 (Table of Contents)

• 1. 서론 1
• 2. 재료 및 방법 3
• 2.1. 피부 섬유아세포 배양과 IL-1β와 IL-10의 처리 3
• 2.2. cDNA probe 정제
• 2.3. RNA 분리와 Northern hybridization 4
• 2.4. Elastin/CAT gene chimeric construc 5
• 2.5. Transfection과 CAT assay 5
• 2.6. Laser confocal microscopy 6
• 2.7. 통계학적 분석 6
• 3. 성적 7
• 3.1. 탄력소 mRNA 발현에 대한 IL-1β의 영향 7
• 3.2. 탄력소 mRNA 발현에 대한 IL-10 의 영향 7
• 3.3. IL-1ß와 IL-10에 의한 탄력소 promoter 유전자의 전사조절 12
• 3.4. IL-1ß와 IL-10 에 의한 단백질 수준에서 탄력소 발현의 증가 12
• 4. 고찰 16
• 5. 요약 20
• 참고문헌 21
• Abstract 25