한약재 추출물 및 대두분획물(LES)이 피부 세포에서 노화관련 유전자 발현에 미치는 영향 및 multiplex-pcr 활용한 효능분석법 개발

다국어 초록 (Multilingual Abstract)

Many phytochemical substances in soy bean were known for effect on anti-cancer, anti-oxidative, anti-aging. Also, extracts from medicinal herbs, Glycyrrhiza uralensis Fischer, Cnidium officinale, Saururus chinensis, Houttuynia cordata have various effects against oxidation and inflammation. To measure expression of genes related to aging, we isolated total RNAs from HEK001 skin cell line treated with LES (Lunasin Enriched Source) and medicinal plant extracts and analyzed a comprehensive gene expression profile of each treated cell. Fifteen genes related aging were selected and confirmed by quantitative real-time RT-PCR analyses. Fifteen genes tested showed higher or lower expression within 2-200 folds in cell line treated with LES and medicinal plant extracts than untreated ones. Also multiplex PCR data shown that we can analysis the effect of medicinal plant. These results imply that these gene profiling can be used to determination of a medicinal materials against aging in skin. Also LES and extract of Cnidium officinale have a role in down regulation of genes related aging, thereby being able to use in products of skin care.

목차 (Table of Contents)

• 목차
• Ⅰ. 서론 1
• Ⅱ. 재료 및 방법 7
• 1. 실험 재료 7
• 2. 실험 방법 8
• 2.1 세포배양 및 추출물 처리 8
• 2.2 Cell viability test 9
• 2.3 Microscopic analysis of morphological change 9
• 2.4 Total RNA preparation 10
• 2.5 cDNA 합성 11
• 2.6 Quantitative RT-PCR 11
• 2.7 Multiplex RT-PCR 12
• 2.8 DNA Gel Electrophoresis 15
• Ⅲ. 결과 및 고찰 16
• 1. 한약재 추출물과 LES가 HEK001 피부세포주의 생존력에 미치는... 16
• 2. LES와 한약재 추출물이 HEK001의 형태적 변화에 미치는 영향 18
• 3. LES와 한약재 추출물이 노화 관련 유전자 발현에 미치는 영향 27
• 4. Multiplex PCR을 이용한 분석법 개발 33
• Ⅳ. 요약 38
• Ⅴ. 참고문헌 40
• Abstract 48