天然物-台灣大戟抗光老化活性成分之研究

余佳純

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天然物-台灣大戟抗光老化活性成分之研究

Anti-photoaging Activity of Natural Products Research in Euphorbia formosana

余佳純 , Masters　　Advisor：李慶國　　Co-advisor ：蕭哲志

繁體中文

抗光老化；基質金屬蛋白酶細胞外基質； anti-photoageing ； matrix metalloproteinases (MMPs) ； extracellular matrix (ECM)

Abstract │ Reference (217) │ Altmetrics

Abstract 〈TOP〉

隨著地球臭氧層的破壞，使得皮膚曝曬在紫外線下的機會增加，導致皮膚提早老化和皮膚癌的發生；在光老化的過程中，基質金屬蛋白酶（Matrix metalloproteinase，簡稱MMPs）扮演了一個重要的角色，本研究藉由抑制 MMP-2 和 MMP-9 的活性，使皮膚細胞外基質（Extracellular matrix，簡稱ECM）不受到 MMP-2 和 MMP-9 的分解而造成皮膚老化。因此，先前本實驗室已將台灣常用的 49 種藥用植物利用 MMP-2 和 MMP-9進行抗光老化的酵素活性篩選，其中以台灣大戟（Euphorbia formosana Hayata）植物抑制 MMP-2 和 MMP-9 的效果為最佳。
    將台灣大戟植物利用人類纖維肉瘤細胞（Human fibrosarcoma cell） HT-1080 細胞中的 MMP-2 與 MMP-9 的活性追蹤方式進行成分的分離及純化，並利用化合物之核磁共振與物理數據等光譜數據，共計純化出 38 個化合物，包括：九個 aliphatic 類化合物、九個 triterpenoid 類化合物、六個 diterpenoid 類化合物、三個 steroid 類化合物、二個 phaeophorbide 類化合物、一個 coumarin 類化合物、一個 aromatic 類化合物、一個 norisoprenoid 類化合物、一個 purine alkaloid 類化合物、一個 polyketide 類化合物、一個 polyprenol 類化合物、一個 lignin 類化合物、二個 others 類化合物。上述38個化合物，其中seco-Helioscopinolde (EF12)、3β,7β-Dihydroxy-ent-abieta-8,13-diene-16,12-olide (EF13) 為新化合物。
    將純化合物對於 MMP-2 和 MMP-9 作活性篩選，實驗結果發現這些化合物對於 MMP-2 與 MMP-9 的抑制效果不佳，與先前的研究結果活性表現不一致，推測是先前的研究中，利用萃取液直接對 MMP-2 和 MMP-9 作酵素反應，而在本研究中則是利用化合物對 HT-1080 細胞作試藥處理，經由細胞內的訊息調控後，取其細胞外液作 Gelatin Zymography 實驗，分析化合物對 MMP-2 與 MMP-9 的活性表現，因此，針對於 MMP 的抑制作用可能藉由不同的基因表現方式，而造成兩者之間對 MMP-2 與 MMP-9 活性表現的差異，期待未來可藉由不同的化合物調控 MMP 的蛋白表現或酵素活性，期待能開發作為新一代天然的抗光老化產品。

Parallel Abstract 〈TOP〉

With increasing UV exposure of skin leads to acute and chronic detrimental cutaneous effects, which may result in development of skin malignancies and photoaging. The Matrix Metalloproteinases (MMPs) play an important role in the photoaging process, which degrade macromolecules of the extracellular matrix (ECM) to increase the happening of wrinkling, sagging and laxity. We examine the MMP-2 and MMP-9 activity for the factor in photoaging process that can degrade collagen and components of the elastic network.
    In the previous studies, our groups had screened 49 species of pharmaceutical plants in Taiwan by anti-photoaging screening model using MMP-2 and MMP-9 enzyme activity. The results demonstrated that Euphorbia formosana had better inhibition of MMPs activity in HT-1080 cells. In Euphorbia formosana, 38 compounds had been isolated and purified by using HPLC and spectroscopy for structure elucidation. There is included nine aliphatic compounds, nine tritepenoids, six diterpenoids, three steroids, two phaeophorbides, one coumarin, one aromatic structure, one norisoprnoid, one purine alkaloid, one polyketide, one poylprenol, one lignan, and the others. Among them, seco-Helioscopinolde (EF12), 3β,7β-Dihydroxy-ent-abieta-8,13-diene-16,12-olide (EF13) are discovered and identified in first time.
    All purified compounds were evaluated the MMP-2 and MMP-9 acitivity in HT-1080 cells, and the results indicated that neither one showed the inhibition of MMP-2 and MMP-9 activity, which were not coincide with the previous study. We were speculating that the difference in crude extracts with MMP-2 and MMP-9 enzyme reaction and compounds were treated to human fibrosarcoma cell (HT-1080) by cell signaling regulation. We supposed that the acitivity of MMP-2 and MMP-9 may be modulated by several genes expression, which resulting in the difference between enzyme reaction and cell signaling. The development for MMP inhibition will require the different compounds in modulating MMP protein expression or enzyme activity, which will be an anti-photoageing products from natural source in new generation.

Reference ( 217 ) 〈TOP〉

5.Goihman-Yahr, M., Skin aging and photoaging: an outlook. Clinical Dermatology 1996, 14, (2), 153-160.
連結：
6.Zhang, G.; Luo, X.; Sumithran, E.; Pua, V.; Barnetson, R.; Halliday, G.; Khachigian, L., Squamous cell carcinoma growth in mice and in culture is regulated by c-Jun and its control of matrix metalloproteinase-2 and -9 expression. Oncogene 2006, 25, 7260-7266.
連結：
8.Urbach, F., The historical aspects of sunscreens. Journal of Photochemistry and Photobiology B: Biology 2001, 64, 99-104.
連結：
9.Lowe, N. J., An Overview of Ultraviolet Radiation, Sunscreens, and Photo-Induced Dermatoses. Dermatologic Clinics 2006, 24, (1), 9-17.
連結：
10.Rosen, C. F., Topical and systemic photoprotection. Dermatologic Therapy 2003, 15, (1), 8-15.
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