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($FSC_{50}$) of extract/fractions of Cayratia japonica were in the order: 50% ethanol extract(114.3 ${\mu}g/mL$)${\mu}g/mL$)${\mu}g/mL$). Reactive oxygen species(ROS) scavenging activities($OSC_{50}$) of some Cayratia japonica extracts in $Fe^{3+}-EDTA/H_2O_2$ system were investigated using the luminol-dependent chemiluminescence assay. The order of ROS scavenging activities were deglycosylated flavonoid aglycone fraction($OSC_{50},\;3.30{\mu}g/mL$)<50% ethanol extract(1.21 ${\mu}g/mL$)${\mu}g/mL$). 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($1{\sim}25{\mu}g/mL$), particularly deglycosylated flavonoid aglycone fraction exhibited the most prominent celluar protective effect(${\tau}_{50}$, 175.05min at 25 ${\mu}g/mL$). 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-${\beta}$-D-glucopyranoside(composition ratio, 11.14%), apigenin-7-O-${\beta}$-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($IC_{50},\;70.5{\mu}g/mL$) 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 $^1O_2$ 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.
본 논문에서는 오렴매 추출물의 항산화, 성분 분석 및 elastase 저해 효과를 알아보았다. 추출물의 free radical(1,1-diphenyl-2-picrylhydrazyl, DPPH) 소거활성($FSC_{50}$)은 50% ethanol 추출물(114.3 ${\mu}g/mL$)$Fe^{3+}-EDTA/H_2O_2$ 계에서 생성된 활성산소종(reactive oxygen species, ROS)에 대한 오렴매 추출물의 총항산화능은 aglycone 분획($OSC_{50},\;3.30{\mu}g/mL$)<50% ethanol 추출물(1.21)$1{\sim}25{\mu}g/mL$)으로 광용혈을 억제하였다. 특히 당을 제거시킨 플라보노이드 aglycone 분획은 25 ${\mu}g/mL$ 농도에서 ${\tau}_{50}$이 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-${\beta}$-D-glucopyranoside, peak 2(15.38%)는 apigenin-7-O-${\beta}$-D-glucuronopyranoside, peak 3(23.55%)는 luteolin, peak 4(49.92%)는 apigenin로 확인되었다. Aglycone 분획은 elastase 저해활성($IC_{50}$)이 70.5 ${\mu}g/mL$로 활성을 나타내었다. 이상의 결과들은 오렴매 추출물이 $^1O_2$ 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.