台灣產葡萄科植物應用於抗光老化化妝品之開發－抑制基質金屬蛋白酶及彈力蛋白酶之探討

溫國慶
中國醫藥大學
皮膚光老化主要係由於紫外線照射誘發活性氧，促使基質金屬蛋白酶-1, 3及9（matrix metalloproteinases, MMP-1, 3, 9）生成增加，降解細胞外基質的膠原蛋白及彈力蛋白，加深皮膚產生皺紋，甚至鬆弛下垂。
多酚類及黃酮類化合物具有抗氧化，清除自由基等作用。由文獻顯示多酚類及黃酮類化合物對纖維母細胞具有抑制基質金屬蛋白酶的作用。葡萄科植物富含多酚類成分，本計畫就下列所載台灣產10種葡萄科植物，台灣產葡萄科植物包括山葡萄(Ampelopsis brevipedunculata)，廣東山葡萄(Ampelopsis cantoniensis)，漢氏山葡萄(Ampelopsis glandulosa)，白蘝(Ampelopsis japonica )，台灣烏歛莓(Cayratia formosana)，虎葛(Cayratia japonica)，粉藤(Cissus repens)，爬牆虎(Parthenocissus dalzielii)，細本葡萄(Vitis thunbergii)，葡萄(Vitis Vinifera)等，探討對MMP-1, 3, 9及彈力蛋白酶的活性。
首先製備葡萄科植物乙醇萃取物及其酸水解物，先進行體外試驗之gelatin digestion test篩選，對於篩選具較佳活性之植物再進行體外細胞試驗包括螢光明膠測試基質金屬蛋白酶活性試驗、彈力蛋白酶（Elastase）活性抑制試驗、UVB 照射實驗、抑制基質金屬蛋白酶-1, 3, 9（MMP-1, 3, 9）活性、第一型膠原蛋白原增生作用及細胞存活率試驗等。接著進行總多酚含量分析，並對具MMP-1, 3, 9活性者再進行其作用機轉之探討。
透過本研究所篩選的葡萄科植物乙醇萃取物及其酸水解物及作用探討之結果，葡萄科植物可期望開發成為抗光老化化妝品。
關鍵字：葡萄科植物、基質金屬蛋白酶、彈力蛋白酶、第一型膠原蛋白原、光老化

Development of Vitaceae plants grown in Taiwan for anti-photoaging cosmetics－The effects on MMPs and elastase

Kuo-Ching Wen
China Medical university
Matrix metalloproteinase-1, 3, 9 (MMP-1, 3, 9) play important roles in photoaging. The production of MMP-1, 3, 9 in skin is increased by UV-irradiation and then cause the degradation of collagen and elastin, even forming coarse wrinkles and sagging skin.
Polyphenols and flavonoids possessed a variety of biological activities including anti-oxidation and free radical scavenging effects. According to the literatures, many Polyphenols and flavonoids had significant inhibitory effects on matrix metalloproteinases in human fibroblasts. In this project, 10 plants of Vitaceae, which are grown in Taiwan, will be selected and applied to investigate the inhibition of MMP-1, 3, 9 and elastase. These plants includes Ampelopsis brevipedunculata, Ampelopsis cantoniensis, Ampelopsis glandulosa, Ampelopsis japonica, Cayratia formosana, Cayratia japonica, Cissus repens, Parthenocissus dalzielii, Vitis thunbergii, and Vitis Vinifera.
At first, the extract of Vitaceae plants and their hydrolysates will be screened by gelatin digestion assay, MMP activity assay using fluorescent gelatin and inhibitory effect on bacterial elastase. Then the inhibitory effect on MMP-1, 3, 9 activities will be further evaluated for the extracts or their hydrolysates passed the screening tests by UV-induced fibroblasts. Secondly, the influences of type I procollagen production and MTT-assay for above extracts or their hydrolysates will be also investigated. In the third, the total phenols content of the extracts or their hydrolysates will be determined. Furthermore, the mechanisms of inhibitory effect on MMP-1, 3, 9 activities for above extracts or their hydrolysates will be investigated through the expression of MAPKs.
Through this study, it could be anticipated that Vitaceae plants could be developed as the effective anti-photoaging agents.
關鍵字：Vitaceae plants, MMPs, elastase, type I procollagen, photoaging