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GDF15 derived from Senescent Fibroblasts Stimulates Melanogenesis in Human Melanocytes
| DC Field | Value | Language |
|---|---|---|
| dc.contributor.advisor | 강희영 | - |
| dc.contributor.author | 김영은 | - |
| dc.date.accessioned | 2019-08-13T16:41:10Z | - |
| dc.date.available | 2019-08-13T16:41:10Z | - |
| dc.date.issued | 2019--8 | - |
| dc.identifier.other | 29327 | - |
| dc.identifier.uri | https://dspace.ajou.ac.kr/handle/2018.oak/15548 | - |
| dc.description | 학위논문(석사)--아주대학교 일반대학원 :의생명과학과,2019. 8 | - |
| dc.description.tableofcontents | ABSTRACT ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ ⅰ TABLE OF CONTENTS ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ ⅱ INTRODUCTION ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 1 Ⅱ. MATERIALS AND METHODS ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 4 1. Cell culture ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 4 2. Enzyme-linked immunosorbent assay (ELISA) ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 5 3. Senescence Associated b-Galactosidase (SA-b-Gal) Staining ‧‧‧‧‧‧‧‧ 5 4. In vitro model of senescent fibroblasts ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 5 5. Ex vivo skin organ culture and pigmentation assay in cultured skin ‧ 6 6. Melanin content and tyrosinase activity assay ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 6 7. Lentivirus and adenovirus production ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 7 8. Semiquantitative real-Time PCR Analysis ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 7 9. Western blot analysis ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 8 10. Immunocytochemistry and immunohistochemical analysis ‧‧‧‧‧‧‧‧‧‧‧ 8 11. Statistical analysis ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 9 Ⅲ. RESULTS ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 10 1. UV induced senescent fibroblasts stimulate skin pigmentation.‧‧‧‧‧‧ 10 2. GDF15 expression is increased in senescent fibroblasts and photo-aged hyperpigmented skin. ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧16 3. GDF15 derived from senescent fibroblasts stimulates melanogenesis in human melanocytes. ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 23 4. GDF15 stimulates b-catenin signaling. ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 35 Ⅳ. DISCUSSION ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 40 Ⅴ. REFERENCES ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 45 국문요약 ‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧‧ 50 | - |
| dc.language.iso | eng | - |
| dc.publisher | The Graduate School, Ajou University | - |
| dc.rights | 아주대학교 논문은 저작권에 의해 보호받습니다. | - |
| dc.title | GDF15 derived from Senescent Fibroblasts Stimulates Melanogenesis in Human Melanocytes | - |
| dc.title.alternative | Yeongeun Kim | - |
| dc.type | Thesis | - |
| dc.contributor.affiliation | 아주대학교 일반대학원 | - |
| dc.contributor.alternativeName | Yeongeun Kim | - |
| dc.contributor.department | 일반대학원 의생명과학과 | - |
| dc.date.awarded | 2019. 8 | - |
| dc.description.degree | Master | - |
| dc.identifier.localId | 952031 | - |
| dc.identifier.uci | I804:41038-000000029327 | - |
| dc.identifier.url | http://dcoll.ajou.ac.kr:9080/dcollection/common/orgView/000000029327 | - |
| dc.description.alternativeAbstract | Background Senescent fibroblasts play a role in aging pigmentation. In our previous RNA sequencing data in senescent fibroblasts, the Growth differentiation factor 15 (GDF-15) gene was found to be significantly upregulated. Objective We investigated the role of senescent fibroblast-derived GDF15 in the regulation of human pigmentation. Methods Normal human melanocytes were co-cultured with fibroblasts infected with GDF15 lenti-virus or sh-GDF15 and melanogenesis was analyzed. The pigmentation were also assessed in the ex vivo skin culture. Results The melanin contents and tyrosinase activity were significantly increased in melanocytes co-cultured with fibroblasts infected with GDF15 lenti-virus. The mRNA and protein expression levels of melanogenesis-associated proteins, microphthalmia-associated transcription factor (MITF) and tyrosinase were significantly up-regulated. The GDF15 stimulated b-catenin signaling in melanocytes. Consistently, the pigmentation were significantly reduced in melanocytes co-cultured with fibroblasts infected with sh-GDF15. The stimulatory action of GDF15 in pigmentation was further confirmed in ex vivo cultured skin. Conclusion Senescent fibroblast-derived GDF15 stimulates skin pigmentation and it may play a role in the aging pigmentation. | - |
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