利用免疫分析法建立皮膚抗氧化體內(in vivo)測試有效性評估之平台

皮膚光老化與氧化壓力所產生的現象有關，而脂肪的過氧化現象(lipid peroxidation)、醣化作用產物(glycation products)及蛋白質的氧化現象(protein oxidation)皆可當作是氧化壓力的生物指標。而其中對於蛋白質的氧化現象，有學者利用免疫轉印呈色方式偵測羰基(carbonyl groups)數量來表示蛋白質被氧化的程度，因為蛋白質羰基可藉由蛋白質的氧化斷裂(oxidative cleavage)或是直接氧化離胺酸、精胺酸、脯胺酸及蘇胺酸殘基而產生，有學者成功利用此生物指標發現在皮膚較外層的角質層(stratum corneum)內的keratin 10被氧化的程度比較內層的角質細胞(keratinocyte)高。因此，我們將利用此偵測蛋白質氧化的方法，設計簡便的免疫分析流程，以建構一套針對抗氧化化妝品具有鑑別度的有效性評估之體內(in vivo)測試平台。由結果顯示我們已找出適當的實驗條件，包括皮屑蛋白質的萃取條件、DNPH(2,4-dinitrophenylhydrazine)與羰基間的反應最佳條件及抗體反應的最佳條件，成功建立具有鑑別度的皮屑蛋白質氧化程度的偵測技術。此技術將可應用在抗氧化化妝品組合配方的抗氧化效力的評估。

關鍵字

蛋白質氧化；抗氧化

並列摘要

Photoaging is known to be correlated with different kinds of oxidative stress. Accumulation of lipid peroxidation, products of glycation, and protein oxidation are often used as biomarkers for oxidative stress. Protein carbonyl may be formed by oxidative cleavage of proteins or direct oxidation of lysine, arginine, praline, and threonine residues. Its level could be detected by immunostainingand used as an indicator for the extent of reactive oxygen-mediated protein oxidation. It have been reported this biomarker have been used for detecting a higher extent of oxidation of human stratum corneum keratin 10 in the outer layer of epidermis (stratum corneum) than in the inner layer of epidermis (keratinocyte). A simple immunoanalysis protocol have been set up based on detecting the extent of protein oxidation. This could facilitate the establishment of an in vivo evaluation platform for anti-oxidative cosmetics. The optimal experiment conditions including extraction of scalp proteins, the reaction of DNPH (2, 4-dinitrophenylhydrazine) with carbonyl, and antibody reactions have been identified. This provides a viable way for detecting the oxidation of scalp proteins. This method could be used in evaluating anti-oxidative cosmetic formulations.

並列關鍵字

protein oxidation ； anti-oxidation

國際替代計量

利用免疫分析法建立皮膚抗氧化體內(in vivo)測試有效性評估之平台

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