Isolation and Purification of cytotoxicity Constituents from Adlay Bran methanolic extract
|Keywords:||薏苡麩皮甲醇萃取物;細胞毒性成分;肺癌細胞;腸癌細胞;adlay bran methanolic extracts;cytotoxicity Constituents;human lung cancer cell A549;colon cancer cell HT-29||Issue Date:||2006||Abstract:||
薏苡（Coix lachryma-jobi L. var. ma-yuen Stapf.）為禾本科一年生草本植物，自古以來即為藥食兼用，傳統醫藥書籍記載其具有除溼利尿、健脾益胃、消炎止痛等功能。目前許多研究報告指出薏苡麩皮甲醇萃出物具抑癌之生理機能，但其主要抗癌活性成分尚未明瞭。本實驗以活性區分為導向來探討薏苡麩皮甲醇萃取物對人類肺癌細胞株 A549 以及腸癌細胞株HT-29 生長之離體效應。以極性大小將薏苡甲醇萃取物依序區分成正己烷、乙酸乙酯、正丁醇層、水層。MTT分析結果發現乙酸乙酯層具有顯著抑制癌細胞之生長能力，故進一步利用管柱層析法及高效能液相層析法等分離技術將乙酸乙酯層分離純化，並以NMR、IR and Mass，鑑定化合物結構。在抗癌活性較佳之乙酸乙酯次區分層(30％-60％ EA/HEX)中分離鑑定出七種化合物。七種化合物中，包植物固醇Sitgmasterol、β-sitosterol、stigmastan-3β-ol、3β-hydroxystigmast-5-en-7-one和酚類化合物3,3’- Dimethoxybiphenyl-4,4’-diol及6-Methoxy-2(3H)-benzoxazolinone，3-O- Caffeoyl-β- sitosterol。
Adlay (Coix lachryma-jobi L. var. ma-yuen Stapf) is a grass crop that has long been used in traditional Chinese medicine and as a nourishing food. According to the folklore of China and Japan, adlay possesses the stomachic, diuretic, antipholgistic, anodynic, antispasmodic, and antitumor effects. A number of recent studies have revealed the physiological mechanism in anti-tumor effects of the adlay bran methanolic extracts, but little is known about its active compounds. Oriented by the separation of active fractions, the aims of this study are to discuss the effects in vitro of adlay bran methanolic extracts on the growth of human lung cancer cell A549 and the colon cancer cell HT-29. Based on the degree of polarity, the adlay bran methanolic extracts are separated to hexane layer, ethyl acetate layer, butanol layer and water layer. The results of MTT analysis showed that the ethyl acetate layer had the significant capacity against growth of cancer cell, so we further isolated and purified the ethyl acetate layer via column chromagraphy and high perferomance liquid chromagraphy to identify their chemical structures by NMR、IR and Mass. Seven chemicals were purified and identified from the ethyl acetate subfraction (30%-60% EA/HEX) which had the better anti-tumor activities. Those of the seven chemicals are sitgmasterol、β-sitosterol、stigmastan-3β-ol、3β-hydroxystigmast-5- en-7-one, 3, 3’-dimethoxybiphenyl-4, 4’-diol, and 6-methoxy-2(3H) - benzoxazolinone,and 3-O-caffeoyl-β-sitosterol.
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