具抗氧化植物萃取液對於自由基誘導DNA 斷裂與細胞死亡之保護效果的評估

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標題:	具抗氧化植物萃取液對於自由基誘導DNA 斷裂與細胞死亡之保護效果的評估 Evaluation of the protective effects of antioxidative herbal extracts on the free radical induced DNA damage and cell death
作者:	江長穎 Chang-Ying Chaing
貢獻者:	林清宮嘉南藥理科技大學：化妝品科技研究所
關鍵字:	植物萃取抗氧化自由基清除皮膚幼? free radical scaven plant extract antioxidant
日期:	2008
上傳時間:	2008-12-29 15:20:11 (UTC+8)
摘要:	在現在化粧品市場中，越來越強調天然化粧品的重要性，除了是因為人們對人工合成化合物有安全疑慮，還有減少廢棄物對於環境傷害等，所以尋找有效的天然物，便成為重要的課題。本研究針對數種的植物萃取液，並以各種評估方法，來驗證其抗氧化的有效性，期望可以應用在化粧品中使用。我們先把蒐集到的植物進行微波萃取，將植物萃取液進行TEAC（ Trolox equivalent antioxidant capacity ）與DPPH 的篩選，再將抗氧化能力較好的萃取液進行0.3%H2O2 經 40mJ/cm2 UV 照射產生 •OH 傷害pUC 119 DNA 的保護試驗，再由DNA 的保護試驗中選出的植物萃取液，以1.2ppm H2O2 與30mJ/cm2 UV 條件下誘導黑色素瘤細胞（B16 melanoma cell）死亡，以不同濃度的植物萃取液進行的保護試驗。最後將植物萃取液添加至乳液中，研究對於毛孔的減少是否有改善。在TEAC 之測定中，由紫蘇葉水萃、山芙蓉葉水萃、荔枝籽水萃、榴槤內皮水萃、榴槤外刺水萃、榴槤內皮乙醇萃有好的抗氧化作用，其 IC50 各為33.93±2.22 、46.11±1.87 、57.04±0.64、72.60±2.89 、72.78±1.33 、94.19±9.00μg/ml。而在清除DPPH‧ 自由基能力之測定，紫蘇葉水萃、刺五加水萃、荔枝籽水萃、榴槤果皮乙醇萃、洛神花酒萃、榴槤外刺水萃有好的抗氧化能力，其IC50 各為 10.61±0.37 、52.51±2.32 、69.05±1.31 、69.79±1.44 、75.27±5.95、84.13±2.84 μg/ml 。所以選出紫蘇葉水萃、刺五加水萃、山芙蓉水萃、荔枝籽水萃、榴槤外刺水萃、榴槤內皮水萃等，做DNA 的保護試驗。而在•OH 使DNA 斷裂的保護試驗中，紫蘇葉水萃、刺五加水萃、山芙蓉水萃、荔枝籽水萃、榴槤外刺水萃、榴槤內皮水萃也都有保護效果，其IC50 各為2.02±0.11、3.38±0.39、3.44±0.33、6.73±0.99、9.22±0.79、9.96±0.40mg/ml。在細胞保護試驗中，在萃取液0.1mg/ml 濃度下，只有山芙蓉葉水萃有可以減少27.6%的細胞死亡，而紫蘇葉水萃並沒有保護能力，反而更增加細胞死亡。山芙蓉葉水萃在0.3mg/ml 下，可以達到減少95.31%的細胞死亡。紫蘇葉在TEAC 與DPPH 中皆擁有非常好的清除能力，而山芙蓉葉只有在TEAC 中可以看見好的清除效果，而兩者在DNA 的保護中，皆有相當好的保護效果。在細胞試驗中，抗氧化能力強的紫蘇葉，產生了細胞毒性，造成細胞死亡，反而是山芙蓉葉水萃，產生了相當好的保護效果。山芙蓉葉水萃在人體上之有效性試驗，塗抹含山芙蓉葉水萃乳液時出現明顯減少受試者的毛孔數，兩週後平均毛孔數降低12.34％。所以我們藉由各種試驗方法的篩選，選出了具化粧品應用潛力之植物，也證實其在人體試確實有效。 In cosmetic market, nature cosmetic products is paid much attention now. Human easy have security anxiety because the manual synthesis compound. Otherwise use nature products can reduce waste to cause the environment pollution. Finding effective nature product is more and more important. Our research is about several plant extract is be estimated to antioxdantive effect. The expectation will be allowed experimental result to use in the cosmetic products. First we carry on the microwave extract with plant. The assays involved different levels of antioxidant action such as radical scavenging abilities using 1,1-diphenyl-2-picryl hydrazyl (DPPH) and trolox equivalent antioxidant capacity (TEAC). In addition, the content of phenols were measured. The better antioxidantive plant extracts were taken to assay protect effect against radiation induced DNA damage in pUC119 plasmid DNA. We induces B16 melanoma cell death in 1.2ppm H2O2 and 30mJ/cm2 UV condition, and than we protect cell death by add different concentration of plant extract. Finally we had application of the emulsion with plant extract to study improvement of the pore reduction.In TEAC assay, Perilla frutescens (L.) Brit. water extract, Hibiscus taiwanensis Hu water extract, Litchi chinensis Sonner. seed water extract, Durio zibethinus Murr.inside rinds water extract, Durio zibethinus Murr. thorn-covered husk water extract and, Durio zibethinus Murr. inside rinds alcohol extract had better activity of scavenged ABTS‧+ free radical (IC50 =33.93±2.22, 46.11±1.87, 57.04±0.64,72.60±2.89, 72.78±1.33, 94.19±9.00 μg/ml, respectively). Perilla frutescens (L.)Brit. water extract, Acanthopanax senticosus(Rupr.et Maxim.)Harmsv water extract,Litchi chinensis Sonner. seed water extract, Durio zibethinus Murr. inside rinds alcohol extract, Hibiscus sabdariffa L alcohol extract, Durio zibethinus Murr.thorn-covered husk water extract scavenged DPPH free radical (IC50=10.61±0.37,52.51±2.32, 69.05±1.31, 69.79±1.44, 75.27±5.95, 84.13±2.84 μg/ml, respectively)have better activity. So we selected Perilla frutescens (L.) Brit. water extract,Acanthopanax senticosus(Rupr.et Maxim.)Harmsv water extract, Hibiscus taiwanensis Hu water extract, Litchi chinensis Sonner. seed water extract, Durio zibethinus Murr. thorn-covered husk water extract, and Durio zibethinus Murr. inside rinds water extract to do DNA protect assay (IC50=2.02±0.11, 3.38±0.39, 3.44±0.33, 6.73±0.99, 9.22±0.79, 9.96±0.40 mg/ml, respectively ). In cell protect assay, Perilla frutescens (L.) Brit. water extract had no protect activity, it induced more cell death. Hibiscus taiwanensis Hu water extract had great activity in free radical induce cell death (protect rare=27.6% in 0.1mg/ml, 95.31% in 0.3mg/ml) .We apply emulsion with Hibiscus taiwanensis Hu water extract to assay efficiency experiment. Our emulsion can decreased 12.34 ％ pore counts in two weeks. Therefore our plant extracts screened by each kind of testing method, we found antioxidative herbal extracts, and also confirmed it in the human body truly effectively.
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