인간각질형성세포에서 Phytol의 항산화 효능
Anti-oxidant Activities of Phytol on Keratinocytes
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목적: 본 연구는 인간각질형성세포에서 phytol의 항산화, 세포보호 효능을 검증하여 화장품 소재로서의 가능성을 알아보 는데 목적이 있다. 방법: Phytol의 효능을 검증하기 위해 세포생존율, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, 2’,7’ -dichlorofluorescin diacetate (DCFDA) assay, reactive oxygen species (ROS) scavenging activity assay, flow cytometry analysis, quantitative real-time polymerase chain reaction (qRT-PCR), glutathione assay를 실시하였다. 결과: 인간각질형성세포에서 H 2 O 2 와 phytol의 실험농도를 결정하기 위해, 다양한 농도로 water-soluble tetrazolium salt (WST-1) assay를 실시하였다. 실험결 과에 따라 H 2 O 2 는 750 µM, phytol은 최대 10 µM을 실험에 사용하였다. 항산화 유전자로 잘 알려진 SOD1, SOD2, CAT mRNA는 phytol 농도 의존적으로 발현이 증가하였고, GSH 또한 phytol 농도 의존적으로 증가하여 항산화 기능이 회복되는 것을 확인하였다. 결론: 이상의 실험들을 통해 phytol의 항산화 효과 및 인간각질형성세포 보호효능을 검증하였으며 잠재적인 노화억제 화장품 소재로서 활용가능성을 제시하였다.
Abstract
Purpose: The aim purpose of this study is to evaluate anti-oxidant and cytoprotective effects of phytol as an ingredient of cosmetic against hydrogen peroxide (H 2 O 2 ) in keratinocytes. Methods: Cell viability assay, 2,2-diphenyl-1-picrylhydrazyl (DPPH) assay, 2’,7’-dichlorofluorescin diacetate (DCFDA) assay, reactive oxygen species (ROS) scavenging activity assay, flow cytometry analysis, quantitative real-time polymerase chain reaction (qRT-PCR), and glutathione assay were performed to verify the cell effectiveness of phytol. Results: To determine the experimental concentration of H 2 O 2 and phytol in HaCaT keratinocytes, water-soluble tetrazolium salt (WST-1) assay was examined with treatments of various H 2 O 2 and phytol concentrations in HaCaT keratinocytes. As a results 750 µM H 2 O 2 and 10 µM phytol treatment was chosen for the experiment. The expression of copper-zinc superoxide dismutase (CuZn-SOD; SOD1 ), manganese superoxide dismutase (Mn-SOD;SOD2), catalase (CAT) mRNA, very well-known as anti-oxidant genes, was increased by phytol in a dose-dependent manner. DPPH radical scavenging activity and intracellular reduced form of glutathione (GSH) levels were increased by phytol in a dose-dependent manner, which showed that anti-oxidant effects are recovered by phytol. Conclusion: It suggests possibility of phytol as a cosmetic ingredient which enhances a defense system and slows an ageing process for skin.
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