藁本成分及其抑制黑色素生成之研究

Studies on Constituents and Melanogenesis-Inhibitory Effects of Ligusticum sinense

朱怡慈(Yi-Tzu Chu)

指導教授 ： 梁文俐

共同指導教授 ： 李慶國

臺北醫學大學/藥學院/生藥學研究所/碩士(2011年)

摘要

中藥「藁本」又稱為「中國的獨活草」，來源是繖形科的 Ligusticum sinense Oliv. 之根莖，其主要功用為散寒解表、袪風勝濕、止痛。而事實上，古籍《神農本草經》亦有記載：藁本具有「長肌膚，悅顏色」之效。本研究的目的是分離鑑定藁本的成分及評估其對於美白的功效。
活性的評估，是藉由體外測定酪胺酸酶的抑制率及 B16 小鼠黑色素瘤细胞的黑色素生成量。藁本甲醇萃取物具有抑制 B16-F10 細胞黑色素生成的效果 (IC50 ≈ 50 μg/mL)，但是不影響酪胺酸酶的酵素活性。經由乙酸乙酯、正丁醇及水的分配萃取後，乙酸乙酯層 (IC50 < 25 μg/mL) 進行開放矽膠管柱層析及高效能液相層析。
藉由活性追蹤的方式進行分離純化，總共鑑定出24個化合物，包含：8個 phenylpropanoids (GB1-8)、6個 phenolics (GB9-14)、2個 phthalide derivatives (GB15-16)、2個 polyacetylenic alcohols (GB17-18)、3個 terpenoids (GB19-21)、1個 steroid (GB22)、1個 fatty acid (GB23)、1個其他類 (GB24)。其中，GB5* 與 GB24* 為新化合物。化合物的結構是利用光譜分析與比對文獻確認。
結果顯示，藁本具有抑制黑色素生成之效。分離到的化合物中，以 GB16 有相對較佳的活性，在沒有細胞毒殺活性的情況下，可抑制 B16-F10 細胞黑色素生成 (IC50 = 31.1 +- 6.8 μM)。

關鍵字

藁本 ； 酪胺酸酶B16-F10黑色素瘤細胞

並列摘要

The rhizoma of Ligusticum sinense Oliv. (Umbelliferae) is the traditional Chinese medicine “Gaoben”, as known as Chinese lovage, mainly used to expel wind-cold and to relieve pain. In fact, Shen Nong’s Herbal, the classic literature of TCM, also noted that Gaoben has a capability to light the skin color. The aims of this study were to isolate the chemical constituents and to evaluate the depigmenting effects of Gaoben.
Anti-melanogenesis properties were determined in vitro by the inhibition of mushroom tyrosinase and the produced melanin content in cultured B16 murine melanoma cells. The methanolic extract of Gaoben showed an inhibitory activity aganist melanogenesis in B16-F10 cells (IC50 ≈ 50 μg/mL), but no effect on tyrosinase activity. Via partitioning with EtOAC, n-Butanol, and H2O, the EtOAC layer (IC50 < 25 μg/mL) was subjected to silica gel open column and HPLC.
Bioassay-guided separation led to the identification of 24 compounds, including eight phenylpropanoids (GB1-8), six phenolics (GB9-14), two phthalide derivatives (GB15-16), two polyacetylenic alcohols (GB17-18), three terpenoids (GB19-21), one steroid (GB22), one fatty acid (GB23) and one other compound (GB24). Among them, GB5* and GB24* are new compounds. Those structures were identified by spectroscopic methods and comparisons with the published data.
The results indiacated that Gaoben showed the definite inhibition of melanin biosynthesis. Among these isolated constituents, GB16 was better than others. It had melanogenesis-inhibitory effect without direct cytotoxicity (IC50 = 31.1 +- 6.8 μM) in B16-F10 cells.

並列關鍵字

Ligusticum sinense ； Tyrosinase ； B16-F10 melanoma cells

參考文獻

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