자외선 A에 의해 유도된 인간각질형성세포 손상에 대한 Scutellarin의 항산화 효과
Anti-oxidant Effects of Scutellarin in UVA-irradiated HaCaT Cells
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목적: 본 연구의 목적은 ultraviolet A (UVA)로부터 산화적 스트레스가 유도된 인간각질형성세포에서 scutellarin의 산화억제 효능을 확인하는데 있다. 방법: Scutellarin의 항산화 효능을 확인하기 위하여 water-soluble tetrazolium salt (WST-1) assay를 이용한 세포독성 측정, 2′,7′-dichlorofluorescin diacetate (DCFDA) assay를 통한 세포 내 활성산소종 정량분석 그리고 quantitative realtime polymerase chain reaction (qRT-PCR)을 통해 superoxide dismutase 1 (SOD1), catalase (CAT) 및 glutathione peroxidase 1 (GPx1) mRNA의 발현 조절 양상을 측정하였다. 결과: Scutellarin는 그 자체로 독성을 보이지 않았고, 농도의존적으로 UVA로부터 세포를 보호하는 효과가 있음이 확인되었다. UVA 조사에서는 세포 내 ROS가 2.15 배 증가하였으나 scutellarin 전처리 군에서는 1.61-1.20 배로 감소하였다. UVA 조사에 의해 저하된 SOD1, CAT 및 GPx1 mRNA의 발현은 scutellarin의 전처리 군에서 농도 의존적으로 증가 하였다. 결론: 본 연구결과를 통하여 scutellarin이 인간각질형성세포에서의 항산화 방어 메커니즘의 조절을 확인 하였고, 향후 scutellarin이 기능성 화장품 개발에 응용될 수 있는 소재로서의 활용을 확인하기 위하여 추가적인 기전 연구가 필요할 것으로 사료된다.
Abstract
Purpose: This study was to investigate the anti-oxidant activity of scutellarin in ultraviolet A (UVA)-irradiated HaCaT cells. Methods: To confirm the anti-oxidant effects of scutellarin, water-soluble tetrazolium salt (WST-1) was used to determine the cytotoxicity, 2′,7′-dichlorofluorescin diacetate (DCFDA) was used to quantitatively analyze intracellular reactive oxygen species (ROS), and quantitative real-time polymerase chain reaction (qRT-PCR) was used to determine the regulation of superoxide dismutase 1 (SOD1), catalase (CAT) and glutathione peroxidase 1 (GPx1) mRNA expression. Results: As a result, scutellarin had no toxicity, and effectively protected HaCaT cells from UVA in a concentration dependent manner. The intracellular ROS was increased by 2.15 times after cell irradiated with UVA, but decreased by 1.61-1.20 times in scutellarin pretreatment group. The expression of SOD1, CAT and GPx1 mRNA, which was downgraded by UVA irradiation, was upregulated in a concentration dependent manner in the scutellarin pretreatment group. Conclusion: This study confirmed that scutellarin regulated anti-oxidant defense mechanisms in HaCaT cells. Future research will be needed to confirm the application of scutellarin as a material for the development of functional cosmetics.
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