인간피부세포에서 복숭아꽃 추출물의 투여가 자외선 B 조사에 의한 활성산소와 Matrix Metalloproteinases 생성 유도를 억제하는 효과
Prevention Effect of Prunus persica Flos Extract from Reactive Oxygen Species Generation and Matrix Metalloproteinases Production Induced by UVB Irradiation in Human Skin Cells
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목적: 복숭아꽃 추출물은 항산화 효과와 항염증 효과가 있다고 보고된 바 있다. 본 연구에서는 복숭아꽃 추출물 투여가 인간피부세포에서 자외선 B 조사에 의한 산화적 스트레스와 matrix metalloproteinase (MMP)의 증가를 억제함으로써 피부 광노화를 예방할 수 있는지 연구하였다. 방법: 복숭아꽃의 에탄올 추출물(Et)과 그로부터 순차적으로 얻은 디클로로메탄(DM), 에틸아세테이트(EA), 부탄올(Bt) 분획을인간 각질형성세포(HaCaT) 또는 인간 피부섬유아세포(HDF)에 전처리하고 자외선 B (75 mJ/cm2)를 조사한 후 활성산소의 생성은 DCF-DA를 이용하여 측정하였고, MMP-1, MMP-3, MMP-9, type-1 procollagen의 mRNA발현과 단백질 합성량은 각각real-time PCR과 Western blot 방법으로 측정하였다. 결과: HaCaT에 복숭아꽃의 Et 추출물과 DM, EA, Bt 분획을 전처리한 결과 모두 자외선 B에 의해 증가한 활성산소와 MMP-1의생성을 유의하게 감소시켰다. 또한, HDF에서 EA와 Bt 분획의 전처리는 자외선 B를 조사한 대조군에 비하여M MP-1, MMP-3, MMP-9 의 mRNA 발현량과 MMP-1과 MMP-3 합성량을 감소시키는 동시에 type-1 procollagen의 mRNA 발현량과 단백질합성량은 증가시켰다. 반면, Et 추출물과 DM 분획은 MMP-1과 MMP-3의 합성은 억제시켰으나y tpe-1 procollagen의 합성을 증가시키지는 못하였다. 결론: 복숭아꽃 에탄올 추출물로부터 얻은 EA와 Bt 분획은 피부세포에서 자외선 B에 의한 산화적 스트레스와 콜라겐의 분해로부터보호하는 효과를 보임으로써 식품 또는 화장품 산업에서 자외선 B에 의해 초래되는 피부 광노화를 예방하는 기능성 제품 개발을 위한천연소재로 이용될 수 있다고 판단된다
Abstract
Purpose: Prunus persica Flos (PPF, peach flowers) extracts were reported to have anti-oxidant and anti-inflammatory activities. In this study, we investigated whether PPF extracts prevent the human skin cells from oxidative stress and matrix metalloproteinases (MMPs) induced by UVB irradiation which ultimately leads to photoaging, or not. Methods: Ethanol extract (Et) of PPF and its dichloromethane (DM), ethylacetate (EA), and n-butanol (Bt) fractions were pretreated in UVB-irradiated (75 mJ/cm2) human keratinocytes (HaCaT) or human dermal fibroblasts (HDF). Cellular reactive oxygen species (ROS) generation was measured using DCF-DA assay, and mRNA expression and production of MMPs were measured by real-time PCR and Western blotting, respectively. Results: The treatment of Et, DM, EA, and Bt significantly inhibited UVB-induced ROS generation and MMP-1 production in HaCaT cells. The treatments of EA and Bt significantly decreased not only the mRNA expression of MMP-1, MMP-3, and MMP-9 but also the protein production of MMP-1 and MMP-3 in HDF cells, while they increased the expression of type-1 procollagen in mRNA and protein levels. Et and DM significantly inhibited only MMP-1 and MMP-3 production without an preventive effect on type-1 procollagen production in UVB-irradiated HDF cells. Conclusion: The treatments with EA and Bt fractions from PPF extract protected human skin cells from UVB-induced oxidative stress and collagen loss. These findings suggest that EA and Bt fractions from PPF extract have the potential of natural resource for a skin anti-photoaging product in the food and cosmetic industry.
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