인간 진피섬유아세포에서의 하이드록시티로졸의 산화억제 효능
Anti-oxidant Effects of Hydroxytyrosol in Human Dermal Fibroblasts
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목적: Hydroxytyrosol은 항산화 특성을 지닌 페놀계 식물 화학 물질의 일종으로, 본 연구에서는 UVA와 hydroxytyrosol을 인간 진피섬유아세포에 처리하여 UVA로 유도된 인간 진피섬유아세포에서의 hydroxytyrosol의 세포 내 작용 기전 연구를 통해 산화 억제에 효과가 있는 기능성 소재로서의 가능성을 제시하고자 한다. 방법: Hydroxytyrosol의 효능 평가는 세포생존율 분석, 활성산소 측정, qRT-PCR을 통해 검증하였다. UVA로 감소한 세포 생존율을 확인하기 위해 WST-1 assay의 원리를 이용하여 측정되었다. Hydroxytyrosol의 항산화 효과를 입증하기 위해서 DCF-DA를 이용하여 ROS 정량분석과 GSH 변화값을 측정하였고, GPx1, SOD1, CAT 유전자의 발현변화를 살펴보았다. 결과: UVA로 감소한 세포생존율은 hydroxytyrosol 농도의존적으로 증가하였다. UVA로 증가된 활성산소에 대해 hydroxytyrosol 처리 시 ROS 소거와 GSH 양의 증가, GPx1, SOD1, CAT mRNA 발현량의 증가를통해 산화 억제 효과를 확인하였다. 결론: 본 연구결과를 통하여 hydroxytyrosol이 인간 진피섬유아세포에서의 산화 억제 효과를확인하였고, 향후 hydroxytyrosol이 적용된 제품을 통한 임상 연구가 진행된다면 항산화제, 화장품의 기능성 소재로써 응용 가능할것으로 판단된다.
Abstract
Purpose: Hydroxytyrosol is a type of phenolic phytochemical with antioxidant properties. In this study, we present the potential of hydroxytyrosol as a functional material with antioxidant effects by determining its intracellular action mechanism in ultraviolet A (UVA)-induced oxidative stress in human dermal firoblasts (HDFs). Methods: Evaluation of the efficacy of hydroxytyrosol was verified through cell survival rate analysis, active oxygen measurement, and quantitative real time polymerase chain reaction (qRT-PCR). The cell survival rate, which was decreased by UVA, increased in a concentration-dependent manner in response to hydroxytyrosol. A WST-1 assay was used to assess the decrease in cell survival rate due to UVA. To verify the antioxidant action of hydroxytyrosol, 2',7'-dichlorofluorescein diacetate (DCF-DA) was used to perform quantitative reactive oxygen species (ROS) analysis and measurement of changes in glutathione (GSH) values, and changes in expression of glutathione peroxidase 1 (GPx1), superoxide dismutase 1 (SOD1) and catalase (CAT) mRNA were also examined. Results: The cell survival rate that was decreased because of UVA treatment was improved in a concentration-dependent manner by hydroxytyrosol. Antioxidant effects of hydroxytyrosol were confimed by the elimination of ROS, increase in the amount of GSH, and increase in the expression levels of GPx1, SOD1, and CAT mRNA. Conclusion: The results of this study suggest that hydroxytyrosol has antioxidant effects in HDFs. We believe that future clinical research on products containing this agent will enable its use in functional materials, such as antioxidants and cosmetics.
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