현초 추출물의 항산화 활성 및 항노화에 관한 연구 = Antioxidative Activities and Antiaging Effects of Geranium nepalense Extracts

국문 초록 (Abstract)

본 연구에서는 현초 추출물의 항산화, 성분 분석 및 elastase와 tyrosinase 저해효과에 관한 실험을 수행하였다. Free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) 소거활성(FSC₅₀)은 50 % 에탄올 추출물(15.0 μg/mL) < ethylacetate 분획(4.5 μg/mL) < 당을 제거시킨 플라보노이드 aglycone 분획(0.9 μg/mL) 순으로 증가하였다. Luminol-의존성 화학발광법을 이용하여 Fe³⁺-EDTA/H₂O₂계에서 생성된 활성산소종(reactive oxygen species, ROS)에 대한 현초 추출물의 총항산화능은 50 % 에탄올 추출물(OSC₅₀, 0.23 μg/mL) < ethylacetate 분획(0.16 μg/mL) < aglycone 분획(0.04 μg/mL)순으로, aglycone 추출물에서 가장 큰 활성을 나타내었다. 현초 추출물에 대하여 rose-bengal로 증감된 사람 적혈구의 광용혈에 대한 억제 효과를 측정하였다. 현초 추출물의 경우 농도 의존적(1 ∼ 50 μg/mL)으로 광용혈을 억제하였다. 특히 당을 제거시킨 플라보노이드 aglycone 분획에서 50 μg/mL 농도에서 τ₅₀이 676.7 min으로 매우 큰 세포보호 효과를 나타내었다. Aglycone 분획은 tyrosinase와 elastase 저해활성(IC₅₀)이 각각 70.0, 19.9 μg/mL로 매우 큰 활성을 나타내었다. 현초 추출물 중 ethylacetate 분획의 당 제거 반응 후 얻어진 aglycone 분획은 TLC에서 2개의 띠로 분리되었으며 HPLC 실험(370 nm)에서도 2개의 피이크로 분리되었다. 분리된 2가지 성분은 quercetin, kaempferol이었으며, 그들의 성분비는 각각 15.3 %, 82.8 %로 kaempferol의 함량이 가장 큰 것으로 나타났다. 이상의 결과들은 현초 추출물이 ¹O₂ 혹은 다른 ROS를 소광시키거나 소거함으로써 그리고 ROS에 대항하여 세포막을 보호함으로써 생체계, 특히 태양 자외선에 노출된 피부에서 항산화제로서 작용할 수 있음을 가리키며, 현초 성분에 대한 분석과 ethylactate분획의 당 제거 실험 후 얻어진 aglycone 분획의 큰 elastase 저해활성으로부터 주름개선 기능성 화장품원료로서도 응용 가능성이 있음을 시사한다.

다국어 초록 (Multilingual Abstract)

In this study, the antioxidative effects and inhibitory effects on elastase and tyrosinase of Geranium nepalense extracts were investigated. The free radical (1,1-diphenyl-2-picrylhydrazyl, DPPH) scavenging activities (FSC₅₀) of extract/fractions of Geranium nepalense were in the order: 50 % ethanol extract (15.0 μg/mL) < ethylacetate fraction (4.5 μg/mL). < deglycosylated flavonoid aglycone fraction (0.9 μg/mL). Reactive oxygen species (ROS) scavenging activities (OSC₅₀) of some Geranium nepalense extracts on ROS generated in Fe³⁺- EDTA/H₂O₂ system were investigated using the luminol-dependent chemiluminescense assay. The order of ROS scavenging activities were 50 % ethanol extract (OSC₅₀, 0.23 μg/mL) < ethylacetate fraction (0.16 μg/mL) < deglycosylated flavonoid fraction (0.04 μg/mL). Deglycosylated flavonoid fraction showed the most prominent scavenging activity. The protective effects of extract/fractions of Geranium nepalense on the rose-bengal sensitized photohemolysis of human erythrocytes were investigated. The Geranium nepalense extracts suppressed photohemolysis in a concentration dependent manner, particularly deglycosylated flavonoid fraction exhibited the most prominent celluar protective effect (τ₅₀, 676.7 min at 50 μg/mL). The inhibitory effect of aglycone fraction on tyrosinase (IC₅₀, 70.0 μg/mL) and elastase (IC₅₀, 19.9 μg/mL) was very high. Aglycone fractions obtained from the deglycosylation reaction of ethyl-acetate fraction among the Geranium nepalense extracts, showed 2 bands in TLC and 2 peaks in HPLC experiments (370 nm). Two components were identified as quercetin (composition ratio, 15.3 %), kaempferol (82.8 %). These results indicate that extract/fractions of Geranium nepalense 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 Geranium nepalense extract and inhibitory activity on elastase of the aglycone fraction could be applicable to new functional cosmetics for smoothing wrinkles.

참고문헌 (Reference)

• 1 S. N. Park, Seoul National Univ. 1989
• 2 박수남, "함초(나문재 및 퉁퉁마디) 추출물의 항산화 작용에 관한 연구(I)" 대한화장품학회 33 (33): 145-152, 2007
• 3 박수남, "제주 자생 식물 추출물의 항산화 및 항노화에 관한 연구(II)" 대한화장품학회 33 (33): 165-173, 2007
• 4 박수남, "쇠뜨기 추출물의 성분 분석과 엘라스타제 활성 저해 효과 연구(Ⅱ)" 대한화장품학회 33 (33): 139-144, 2007
• 5 G. F. Vile, "UVA radiation-in duced oxidative damage to lipid and protein in vitro and in human skin fibroblasts is dependent on iron and singlet oxygen" 18 : 721-, 1995
• 6 L. H. Kligman, "UVA induced biochemical changes in hairless mouse skin collagen: a contrast to UVB effects" 209 : 1992
• 7 S. H. Kim, "The effect of kaempferol, quercetin on hyaluronan synthesis stimulation in human keratinocytes (HaCaT)" 33 (33): 97-, 2005
• 8 S. N. Park, "Skin aging and antioxidant" 23 : 75-, 1997
• 9 M. Wlaschek, "Singlet oxygen may mediate the ultraviolet A in induced synthesis of interstitial collagenase" 104 : 194-, 1995
• 10 R. M. Tyrrell, "Singlet oxygen involvement in the inactivation of cultured human fibroblast by UVA and near visible radiations" 49 : 407-, 1989
• 11 K. Scharffetter-Kochanek, "Singlet oxygen induces collagenase expression in human skin fibroblasts" 331 : 1993
• 12 S. N. Park, "S. N. Park, Antioxidative properties of baicalein, component from Scutellaria baicalensis Georgi and its application to cosmetics (I)" 14 (14): 657-, 2003
• 13 J. C. Fantone, "Role of oxygen-derived free radicals and metabolites in leukocyte dependent inflammatory reaction" 107 : 397-, 1982
• 14 K. J. A. Davies, "Protein damage and degradation by oxygen radical" 262 : 9895-, 1987
• 15 C. S. Foote, "Photosensitized oxidation and singlet oxygen; consequences in biological systems" Acdemic press 2 : 85-, 1976
• 16 K. Scharffetter-Kochanek, "Photoaging of the connective tissue of skin: its prevention and therapy, antioxidants in disease mechanism and therapy" 38 : 639-, 1997
• 17 박수남, "O2으로 유도된 사람 적혈구의 광용혈에 있어서 대두의 아이소플라본인 제니스테인의 보호작용" 한국식품과학회 35 (35): 510-518, 2003
• 18 R. Fleischmajor, "Human dermal glucosaminoglycans and aging" 279 (279): 265-, 1972
• 19 M. O. Longas, "Evidence for structural changes in dermatan sulfate and hyaluronic acid with aging" 159 (159): 127-, 1987
• 20 H. J. Yang, "Evaluation of antioxidant potential of extract/fractions of Equisetum arense (Ⅰ)" 33 (33): 61-, 2007
• 21 M. H. Jung, "Effect of Geranii herba water extract on gastric secretion and experimental ulceration in rats" 10 (10): 1-, 1980
• 22 Y. H. Choi, "Dyeability and antibacterial activity of the fabrics with Elm-bark extracts" 15 : 140-, 2003
• 23 J. H. Kim, "Dyeability and anti- bacterial activity of fabrics using balsamine extracts" 15 : 15-, 2003
• 24 A. Oikarinen, "Connective tissue alterations in skin exposed to natural and therapeutic UV-radiation" 2 : 15-, 1985
• 25 J. W. Choi, "Antioxidative and cellular protective effects of Jeju plant extracts against reactive oxygen species (Ⅰ)" 32 (32): 181-, 2006
• 26 K. J. Yong, "Antibacterial and deordorization activities of cotton fabrics by amur cork tree extracts" 11 : 9-, 1999
• 27 A. Oikarinen, "A biochemical and immunohistochemical study of collagen in sun-exposed and protected skin" 6 : 24-, 1989