以不同細胞模式評估闊葉大豆萃取物之植物雌激素活性

Evaluation of the phytoestrogenic activity of Glycine tomentella extracts by different cellular models

林娟娟(Chuan-Chuan Lin) ； 王經強(Ching-Chiang Wang) ； 劉滿海(Man-Hai Liu)

《中華科技大學學報》 79期 (2020/05) Pp. 135-152

摘要

闊葉大豆（Glycine tomentella）為台灣金門特有豆科一條根品種，富含具有功效的植物雌激素異黃酮類及酚類物質，特別是異黃酮類的genistein和daidzein，具有抗氧化、消炎、降血脂、免疫調節及預防骨質疏鬆之功效。本研究使用管柱層析法分別從金門一條根GT（Glycine tomentella）及比較其他市售大陸一條根FL（Flemingia species）中以不同比例正己烷／乙酸乙酯沖提液製備不同極性區間萃取物，檢測其抗氧化、多酚及主要異黃酮成分含量分析，再分別以基因工程建構之酵母菌及小鼠骨母細胞培養平台篩選比較，試圖由市售一條根中找出最具植物雌激素活性之品種。結果發現將GT與FL萃取物對骨母細胞增生之作用和先前以酵母菌篩選雌激素活性及HPLC檢測含量結果做比較，GT比FL萃取物於酵母菌系統中具雌激素活性，也有較好的骨母細胞增生作用，由兩種篩選系統結果皆顯示genistein 活性遠大於daidzein，所以同時含有genistein及daidzein之GT-2（Hex：EA=1：1 fraction）於酵母菌及小鼠骨母細胞系統中均較GT-3（Hex：EA=1：3 fraction）具活性雌激素及骨母細胞增生之作用，也為最佳萃取物作為開發成保健產品，以取代雌激素為預防更年期婦女骨質疏鬆之用途。

關鍵字

異黃酮 ； 抗氧化 ； 植物雌激素 ； 骨母細胞模式

並列摘要

I-Tiao-Gung, including Glycine tomentella and Flemingia species, possesses several functional phytoestrogens and bioflavonoids, especially genistein and daidzein, which have been reported to have anti-oxidative, anti-inflammatory, hypolipidemic, immunomodulatory effects as well as prevention of osteoporosis. In our study, several fractions after column chromatography from commercially available species, Glycine tomentella and Flemingia species, were prepared by using different ratios of hexane/ethyl acetate as eluents. First, the antioxidant activity, the contents of polyphenols and isoflavonoids were analyzed and compared. In order to find the most phytoestrogens-enriched species, a recombinant yeast-based screening assay and a mouse osteoblast cellular model were used in this study. The results indicated that GT extract possessed much more photoestrognic activity than FL extract by comparing the results among the yeast-based screening assay, HPLC analysis of the contents of major isoflavonoids and the proliferation of osteoblast. In addition, the two screening systems both indicated that genistein exhibited greater bioactivity than daizein. Therefore, it is expected that GT-2 (Hex : EA=1 : 1 fraction), which contains both genistein and daidzein, exhibited greater phytoestrogenic activity than the GT-3 (Hex：EA=1：3 fraction) containing only daidzein, as shown in both yeast assay and the proliferation of mouse osteoblast. The study also demonstrated that the GT-2 extract after the optimized process of fractionation would be an alternative choice for the development of functional product for preventing menopausal osteoporosis.

並列關鍵字

isoflavonoid ； antioxidant ； phytoestrogen ； osteoblast model