산거울 추출물의 Elastase 활성 저해 및 Matrix Metalloproteinase-1 발현 억제 효과 = Inhibitory Effects of Carex humilis Extract on Elastase Activity and Matrix Metalloproteinase-1 Expression

국문 초록 (Abstract)

본 연구에서는 산거울 추출물의 항주름 활성을 연구하기 위하여 항산화, 엘라스타제 활성 억제, Matrix metalloproteinase-1 (MMP-1) mRNA 발현 및 단백질의 생성 억제 관련 실험을 진행하였다. 산거울의 뿌리부분을 95 % 에탄올로 추출하고 유기 용매로 분획하여 각 추출물 분획물에 대한 항산화 활성과 엘라스타제 억제 효능을 측정한 결과, EtOAc 분획이 각각 SC50=4.89 μg/mL, IC50=23.5 μg/mL 로 가장 우수한 결과를 나타내었다. 따라서 활성이 가장 좋은 EtOAc 분획물로 부터 실리카겔 컬럼 크로마토그래피를 이용하여 활성물질을 분리하고, 분리된 물질이 α-viniferin임을 분광학적 분석결과로 확인하였다. RT-PCR을 이용한 MMP-1 mRNA 발현능 평가에서 EtOAc 분획물과 α-viniferin은 각각 50 ~ 60 % (10 ~ 100 μg/mL), 약 60 ~ 75 % (0.5 ~ 2 μg/mL)의 저해효과를 나타내었으며, Western-blot을 이용한 MMP-1 단백질 생성을 평가한 결과에서 역시 EtOAc 분획물과 α-viniferin은 같은 농도에서 각각 50 ~ 65 %와 55 ~ 65 % 저해 효과를 나타내었다. 결론적으로, α-viniferin을 함유한 산거울의 EtOAc 분획층의 주름 개선용 기능성 화장품 개발을 위한 소재로 개발 가능성을 확인하였다.

다국어 초록 (Multilingual Abstract)

In order to evaluate anti-wrinkle activity of Carex humilis extract, free radical scavenging activity, elastase inhibitory activity and reduction of expression Matrix metalloproteinase-1 (MMP-1) mRNA and MMP-1 protein were investigated. The roots of Carex humilis were extracted with 95 % ethanol and successively partitioned with organic solvents with increasing polarity of the solvents. Each fraction of organic solvent were investigated by using free radical scavenging activity and elastase inhibitory activity test. Among them, EtOAc fraction showed antioxidant activity (SC₅₀=4.89 μg/mL) and elastase inhibitory activity (IC₅₀=23.5 μg/mL). EtOAc fraction was developed on silica gel by open-column chromatography and consecutively re-developed on C18 resin by prep-HPLC to give α-viniferin as a major component, which was confirmed by spectrometric analysis. In the assay on expression of MMP-1 mRNA by RT-PCR and protein by western-blot, EtOAc layer (10 ~ 100 μg/mL) was reduced about 50 ~ 60 %, 50 ~ 65 % respectively and α-viniferin (0.5 ~ 2 μg/mL) was inhibited about 60 ~ 75 %, 55 ~ 65 % respectively in human fibroblast. Therefore, our findings suggest that EtOAc layer of Carex humilis containing α- viniferin can be useful as an active ingredient for cosmeceuticals of anti-wrinkle effects.

참고문헌 (Reference)

• 1 이영노, "한국식물도감" 교학사 1086-, 1998
• 2 R. J. Pryce, "α-viniferin: an antifungal resveratrol trimer from grapevines" 18 (18): 213-, 1981
• 3 S. H Lee, "α-Viniferin: A Prostaglandin H2 Synthase Inhibitor from Root of Carex humilis" 64 (64): 204-, 1998
• 4 P. Brenneisen, "Ultraviolet B Wavelength Dependence for the Regulation of Two Major Matrix-Metalloproteinases and Their Inhibitor TIMP-1 in Human Dermal Fibroblasts" 64 (64): 877-, 1996
• 5 D. R. Keene, "Type VII collagen forms an extended network of anchoring fibrils" 104 : 611-, 1987
• 6 S. A. Chowdhury, "Tumor-specificity and apoptosis-inducing activity of stilben and flavonoids" 25 (25): 2055-, 2005
• 7 K. Fraizer, "Stimulation of fibroblast cell-growth, matrix production, and granulation-tissue formation by connective-tissue growth-factor" 107 (107): 404-, 1996
• 8 J. S. Choi, "Screening on radical savenging activity of marine microalgae" 21 (21): 252-, 2000
• 9 B. H. Malgorzata, "Resveratrol oligomer are potent MRP1 tracsport inhibitiors" 26 (26): 2081-, 2006
• 10 T. Peter, "Physiology Of The Skin, Allured Publishing Corporation" 1 : 61-, 2001
• 11 K. Takeda, "Funtional Cosmetology, society of cosmetic chemists of Japan" 1 : 163-, 2004
• 12 H. H. Euis, "Cytotoxic resveratrol oligomers from the tree bark of Dipterocarpus hasseltii" 77 : 550-, 2006
• 13 A. A. Van de Loosdrecht, "Cell mediated cytotoxicity against U 937 cells by human monocytes and macrophages in a modified colorimetric MTT assay" 141 (141): 15-, 1991
• 14 E. Y. Chung, "Anti-inflammatory effect of the oligometric stilbene alphaviniferin and its mode of the action though inhibition of cyclooxygenase-2 and inducible nitric oxide synthase" 69 (69): 710-, 2003
• 15 S. Kitanaka, "(+)α- Viniferin, an Anti-inflammatory Compound from Caragana Chamlagu Root" 38 : 432-, 1990
• 16 S. H. Sung, "(+)-α-Viniferin, a Stilbene Trimer from Caragana chamlague, Inhibits Acetylcholinesterase" 25 (25): 125-, 2002
• 17 S. Kitanaka, "(+)-a-Viniferin, an anti-inflammatory compound from Caragana chamlagu root" 38 (38): 432-, 1990
• 18 S. H. Sung, "(+)-Alphaviniferin, a stilbene Trimer from Caragana Chamlague, inhibits acetylcholinesterase" 25 (25): 125-, 2002