오렴매 추출물의 항산화 활성, 성분 분석 = Antioxidative Activity and Component Analysis of Cayratia japonica Extract

국문 초록 (Abstract)

본 논문에서는 오렴매 추출물의 항산화, 성분 분석 및 elastase 저해 효과를 알아보았다. 추출물의 free radical(1,1-diphenyl-2-picrylhydrazyl, DPPH) 소거활성(FSC₅₀)은 50 % ethanol 추출물(114.3 μg/mL) < ethyl acetate 분획(59.4) < 당을 제거시킨 플라보노이드 aglycone 분획(58.8) 순으로 증가하였다. Luminol-의존성 화학발광법을 이용한 Fe³⁺-EDTA/H₂O₂ 계에서 생성된 활성산소종(reactive oxygen species, ROS)에 대한 오렴매 추출물의 총항산화능은 aglycone 분획(OSC₅₀, 3.30 μg/mL) < 50 % ethanol 추출물(1.21) < ethyl acetate 분획(0.34)순으로, ethyl acetate 분획에서 가장 큰 활성을 나타내었다. 오렴매 추출물에 대하여 rose-bengal로 증감된 사람 적혈구의 광용혈에 대한 억제 효과를 측정하였다. 오렴매 추출물의 경우 농도 의존적(1 ~ 25 μg/mL)으로 광용혈을 억제하였다. 특히 당을 제거시킨 플라보노이드 aglycone 분획은 25 μg/mL 농도에서 τ₅₀이 175.05 min으로 매우 큰 세포보호 효과를 나타내었다. 오렴매 추출물 중 ethyl acetate 분획의 당 제거 반응 후 얻어진 aglycone 분획은 TLC에서 2개의 띠로 분리되었으며, HPLC 실험(360 nm)에서 2개의 피이크로 분리되었다. 분리된 2가지 성분은 luteolin, apigenin이었으며, 그들의 성분비는 각각 47.50 %, 52.50 %로 나타났다. 오렴매 추출물의 ethyl acetate 분획의 TLC 크로마토그램은 3개의 띠로 분리되었고, HPLC 크로마토그램은 4개의 피이크를 보여주었다. TLC와 HPLC의 띠와 피이크를 확인한 결과, HPLC의 4개의 피이크는 용리순서로 peak 1 (조성비 11.14 %)은 luteolin-7-O-β-D-glucopyranoside, peak 2 (15.38 %)는 apigenin-7-O-β-D-glucuronopyranoside, peak 3 (23.55 %)는 luteolin, peak 4 (49.92 %)는 apigenin로 확인되었다. Aglycone 분획은 elastase 저해활성(IC₅₀)이 70.5 μg/mL로 활성을 나타내었다. 이상의 결과들은 오렴매 추출물이 ¹O₂ 혹은 다른 ROS를 소광시키거나 소거함으로써 그리고 ROS에 대항하여 세포막을 보호함으로써 생체계, 특히 태양 자외선에 노출된 피부에서 항산화제로서 작용할 수 있음을 가리키며, 성분 분석과 이와 같은 항산화 활성을 통하여 화장품 원료로서 응용 가능성이 있음을 시사한다.

다국어 초록 (Multilingual Abstract)

In this study, the antioxidative effects, inhibitory effects on elastase, and components of Cayratia japonica extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities (FSC₅₀) of extract/fractions of Cayratia japonica were in the order: 50 % ethanol extract (114.3 μg/mL) < ethyl acetate fraction (59.4 μg/mL) < deglycosylated flavonoid aglycone fraction (58.8 μg/mL). Reactive oxygen species (ROS) scavenging activities (OSC₅₀) of some Cayratia japonica extracts in Fe³⁺-EDTA/H₂O₂ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activities were deglycosylated flavonoid aglycone fraction (OSC₅₀, 3.30 μg/mL) < 50 % ethanol extract (1.21 μg/mL)< ethyl acetate fraction (0.34 μg/mL). Ethyl acetate fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Cayratia japonica on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Cayratia japonica extracts suppressed photohemolysis in a concentration dependent manner (1 ~ 25 μg/mL), particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect (τ₅₀, 175.05 min at 25 μg/mL). Aglycone fractions obtained from the deglycosylation reaction of ethyl acetate fraction among the Cayratia japonica extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments (360 nm). Two components were identified as luteolin (composition ratio, 47.50 %), apigenin (52.50). TLC chromatogram of ethyl acetate fraction of Cayratia japonica extract revealed 3 bands and HPLC chromatogram showed 4 peaks, which were identified as luteolin-7-O-β-D-glucopyranoside (composition ratio, 11.14 %), apigenin-7-O-β-D-glucuronopyranoside (15.38 %), luteolin (23.55 %) and apigenin (49.92 %) in the order of elution time. The inhibitory effect of aglycone fraction on elastase (IC₅₀, 70.5 μg/mL) was very high. These results indicate that extract/fractions of Cayratia japonica can function as antioxidants in biological systems, particularly skin exposed to UV radiation by scavenging ¹O₂ and other ROS, and protect cellular membranes against ROS. And component analysis of Cayratia japonica extract and antioxidative effects could be applicable to new cosmetics.

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