白蘝與蜜紅葡萄及其成分之製劑製備暨其抗光老化及抗光致癌性機制之探討
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的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。皮膚光致癌性也因紫外線誘發之活性氧使皮膚脂質產生脂質烷氧自由基（lipid alkoxy radicals）與細胞中之DNA結合而引發。同時DNA亦會吸收UVB的能量而誘發光產物如環丁烷嘧啶雙體（cyclobutane pyrimidine dimmers, CPDs）而誘發哺乳細胞突變。
就先前研究結果顯示，葡萄科植物中以白蘝與蜜紅葡萄具有較強之抑制MMPs作用，本計畫擬選定白蘝與蜜紅葡萄及其主要成分楊梅黃酮（myricetin）與槲皮素（quercetin）為標的，探討其抑制MMPs活性及光致癌性之上游調控機制，並進行上述標的之固態脂質奈米粒（Solid lipid nanoparticles, SLN）及奈米結構脂質載體（nanostructured lipid carrier, NLC）配方之探討，以促進其吸收並做為其效用之實證依據。
本計畫第一年擬進行楊梅黃酮與槲皮素及白蘝與蜜紅葡萄萃取物等抑制MMPs活性之上游調控機制之探討，包括纖維母細胞經UVB照射後MAPKs (JNK, ERK, p-38), AP-1 (c-Jun, c-Fos) 及S-mad等之表現。以及楊梅黃酮與槲皮素之傳統乳劑，SLN及兩種NLC配方（NLC-A, NLC-O/F/W）製作，並探討四種劑型之經皮吸收效果。
第二年擬進行楊梅黃酮與槲皮素及白蘝與蜜紅葡萄萃取物等對光致癌性之保護及其對調控NF-kB/p65之表現。以及就第一年所得最適宜的楊梅黃酮與槲皮素之SLN、NLC-A, NLC-O/F/W配方，製作白蘝與蜜紅葡萄萃取物之傳統乳劑，SLN、NLC-A, NLC-O/F/W配方，以楊梅黃酮與槲皮素成分為指標，探討四種劑型之經皮吸收效果。
透過本研究製作新劑型探討之結果，楊梅黃酮與槲皮素及白蘝與蜜紅葡萄萃取物等可期望開發成為抗光老化及光致癌性保護之產品。
關鍵字：楊梅黃酮，槲皮素，白蘝，蜜紅葡萄，固態脂質奈米粒，奈米結構脂質載體
The investigation on preparations of Ampelopsis japonica, Vitis vinifera x-Vitis labrus Cal. and their active constituents and their mechanisms for anti-photoaging and anti-photocarcinogenesis
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. The increase then causes the degradation of collagen and elastin even forms coarse wrinkles and sagging skin. Skin photocarcinogenecity also causes by UV-induced ROS, which can initiate lipid peroxidation and produce lipid alkoxy radicals, and then combine with DNA to form DNA adducts. The energy of UVB is absorbed by DNA and induces photoproducts such as cyclobutane pyrimidine dimers (CPDs), which contribute to mutation in mammalian cells.
In our previous study, Ampelopsis japonica and Vitis vinifera x-Vitis labrus showed the inhibition of UVB-induced MMPs expression. In this study, Ampelopsis japonica, Vitis vinifera x-Vitis labrus, myricetin and quercetin were subjected to investigate the mechanisms of MMPs inhibition and anti-photocarcinogenesis. On the other hand, the new dosage forms, solid lipid nanoparticles (SLN) and nanostructured lipid carrier (NLC) of the above mentioned components will be prepared.
In the first year, the mechanisms of MMPs inhibition of Ampelopsis japonica, Vitis vinifera x-Vitis labrus, myricetin and quercetin, including the expression of MAPKs (JNK, ERK, p-38), AP-1 (c-Jun, c-Fos) and S-mad, will be investigated in fibroblasts after UVB exposure. On the other hand, the traditional cream, SLN, NLC-A and NLC-O/F/W of myricetin and quercetin will be prepared for evaluating the percutaneous absorption.
In the second year, the effect of anti-photocarcinogenesis and its mechanism, the regulation of NF-kB/p65 expression of Ampelopsis japonica, Vitis Vinifera x-Vitis labrus, myricetin and quercetin after UVB exposure will be investigated. On the other hand, the optimized formulations of myricetin and quercetin SLN, NLC-A and NLC-O/F/W which obtained through the evaluation in the first year will be adopted for preparing SLN, NLC-A and NLC-O/F/W of Ampelopsis japonica, Vitis vinifera x-Vitis labrus. Furthermore, myricetin and quercetin as the marker substances will be evaluated for the percutaneous absorption on the traditional cream, SLN, NLC-A and NLC-O/F/W of Ampelopsis japonica, Vitis vinifera x-Vitis labrus.
This study would be a pilot study for developing the new dosage forms of Ampelopsis japonica, Vitis vinifera x-Vitis labrus, myricetin and quercetin for anti-photoaging and photocarcinogenic protection.
關鍵字：myricetin, quercetin, Ampelopsis japonica, Vitis vinifera x-Vitis labrus, solid lipid nanoparticles, nanostructured lipid carrier
