| 摘要: | 骨質疏鬆症 (Osteoporosis) 好發於女性，其肇因與婦女停經後缺乏雌激素及老化有關。臨床上，雌激素為停經後婦女常用之抗骨質疏鬆藥物，但其副作用有增加子宮內膜癌和乳癌發生的機會，而天然物未開發的領域尚多，尚可從其開發出更具潛力之藥物。本實驗室過去曾進行兔尾草 95% 乙醇萃取物研究，發現其於人類初代造骨細胞具有造骨活性，因此本論文擬進一步以體內及體外試驗探討兔尾草水萃物之造骨活性，並進行其化學成分分離。兔尾草根部以熱水進行萃取，將水萃物投予經卵巢切除術之 Sprague-Dawley (SD) 大鼠，分別投予低、高劑量兔尾草水萃物、雌性激素、福善美，共餵食90天；兔尾草水萃物利用多種層析管柱並配合人類初代造骨細胞 (HOb cell) 以活性追蹤方式進行成分分離，並以造骨指標分析其活性。結果顯示，於體內試驗投予兔尾草水萃物各組均無肝腎功能異常；生物力學檢測顯示投予兔尾草水萃物低劑量及高劑量組均能增加骨硬度 (Stiffness); 微電腦斷層掃描顯示投予兔尾草水萃物高劑量組能增加椎骨之骨密度 (BMD)、骨體積與組織的比率 (BV/TV)、骨小樑數目 (Tb.N)、降低骨小樑分離度 (Tb.Sp)。於體內試驗發現其具有造骨活性，因此進一步配合體外試驗以活性追蹤方式分離兔尾草之化學成分，得 UCH-1 及新化合物 UCH-2，於體外試驗發現兩化合物均可促進鹼性磷酸酶之分泌，且 UCH-2 可促進礦化活性，為其活性指標成分。綜合上述結果，兔尾草可提供未來開發為抗骨質疏鬆症藥物之參考。
Menopausal hormone decline and aging contributes to increase the risk of osteoporosis. In clinical treatments, although estrogen therapy has been a widely used to prevent or treat post-menopausal osteoporosis, it will increase the risk of breast and uterine cancer and other undesirable side effects. The field of natural product is still much untapped, from which it expects to develop a better antiosteoporosis drug. It had been investiggted in our laboratory that the 95% ethanol extracts of Uraria crinita exhibited osteogenic activities in human osteoblasts (HOb cells) in vitro. In this study, the in vivo and in vitro experiments of hot water extracts from U. crinita, and isolated active components were investigated. We extracted the roots of U. crinita with aqueous, and administered low-dose U. crinita aqueous extract, high-dose U. crinita aqueous extract, 17β-estradiol and alendronate sodium trihydrate to ovariectomized Sprague-Dawley (SD) rats respectively. Each group contains ten rats and treated for 90 days. U. crinita aqueous extract was further isolated and purified with variety of columns. We obtained two compounds, and used alkaline phosphatase methods to detect its osteogenic activities. Results showed both two groups which administrated U. crinita aqueous extract had no liver and kidney dysfunction; biomechanical analysis showed that administrated U. crinita aqueous extract increased bone stiffness significantly; the parameter of microcomputer tomography show that the group which administrated high-dose U. crinita aqueous extracts increased bone mineral density (BMD), bone volume /tissue volume (BV/TV), trabecular number (Tb.N) and reduced trabecular separation (Tb.Sp) in vertebrae significantly. After the bio-guided column chromatography, we obtained two compounds UCH-1 and UCH-2. These two compounds increased the ALP activity significantly, and UCH-2 increased the mineralization activity significantly. Based on the results, U. crinita is a potential source to be developed into anti-osteoporosis drugs, and provide a reference for anti-osteoporosis drug development. |
