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RESEARCH ARTICLE

Anti-oxidant Activities of Phytol on Keratinocytes

원문정보

인간각질형성세포에서 Phytol의 항산화 효능

Sun-Hee Jeong

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초록

영어

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 (H2O2) 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 H2O2 and phytol in HaCaT keratinocytes, water-soluble tetrazolium salt (WST-1) assay was examined with treatments of various H2O2 and phytol concentrations in HaCaT keratinocytes. As a results 750 μM H2O2 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.

한국어

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 H2O2와 phytol의 실험농도를 결정하기 위해, 다양한 농도로 water-soluble tetrazolium salt (WST-1) assay를 실시하였다. 실험결 과에 따라 H2O2는 750 μM, phytol은 최대 10 μM을 실험에 사용하였다. 항산화 유전자로 잘 알려진 SOD1, SOD2, CAT mRNA는 phytol 농도 의존적으로 발현이 증가하였고, GSH 또한 p hytol 농도 의존적으로 증가하여 항산화 기능이 회복되는 것을 확인하였다. 결론: 이상의 실험들을 통해 phytol의 항산화 효과 및 인간각질형성세포 보호효능을 검증하였으며 잠재적인 노화억제 화장품 소재로 서 활용가능성을 제시하였다.

중국어

目的: 鉴定人角质形成细胞中phytol的抗氧化和细胞保护效能,探索作为化妆品原料的可行性。方法: 为确定phytol的效 能,进行细胞生存率,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 assay等实验。结果: 在人角质形成细胞中,为确定H2O2和phytol的实验浓度,对 不同浓度的phytol进行water-soluble tetrazolium salt (WST-1) assay。实验结果显示, H2O2的最佳实验浓度为750 μM, phytol的最大范围使用浓度为10 μM。抗氧化遗传因子SOD1、SOD2、CAT mRNA的表达程度根据浓度依赖性逐渐增加, GSH也根据浓度依赖性逐渐恢复其抗氧化能力。结论: 通过以上的实验结果,鉴定了phytol的抗氧化效能以及人角质形成 细胞的保护效果,因此作为潜在的抗衰老化妆品原料充分提出了活用可行性。

목차

Abstract
 Introduction
 Methods
  1. 세포배양
  2. 세포생존율 측정
  3. 세포보호 효과 측정
  4. DPPH 라디칼 소거 활성 측정
  5. 세포 내 ROS 정량 분석
  6. 항산화 관련 유전자의 발현 분석
  7. GSH 측정
  8. 통계처리
 Results and Discussion
  1. 인간각질형성세포에서 H2O2에 의한 세포독성
  2. H2O2에 대한 phytol의 세포보호 효과
  3. Phytol의 DPPH 라디칼 소거 활성
  4. Phytol의 세포 내 ROS 억제 활성
  5. Phytol의 항산화 유전자 발현 촉진 효과
 Conclusion
 Acknowledgements
 References
 국문초록
 中文摘要

저자정보

  • Sun-Hee Jeong 정선희. Department of Beauty Arts, Suwon Women’s University, Suwon-si, Gyeonggi-do, Korea

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