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    題名: 探討大豆異黃酮素對於老鼠造骨細胞增生與骨蛋白表現的影響
    The Effects of Soy Isoflavones ( Genistein and Daidzein) on MC3T3-E1 Cells Proliferation and Bone Protein Expression
    作者: 郭瑋綺
    Wei-Chi Kuo
    貢獻者: 保健營養學研究所
    關鍵詞: Genistein
    Daidzein
    MC3T3-E1細胞
    Osteopontin
    第一型膠原蛋白
    鹼性磷酸酶
    Genistein
    Daidzein
    MC3T3-E1 cells
    Osteopontin
    Type I collagen
    Alkaline phosphatase
    日期: 2004
    上傳時間: 2009-09-10 18:03:43 (UTC+8)
    摘要: 本研究針對大豆異黃酮素(Genistein、Daidzein)對於老鼠造骨細胞株(MC3T3-E1 cells)增生之影響機制進行探討。由細胞增生實驗結果(MTS assay)發現投與10-9M~10-5M濃度的異黃酮素，培養24小時，與控制組比較，可顯著刺激MC3T3-E1細胞的增生(p<0.05)；培養24~96小時，計算細胞增生數目，發現10-9M~10-6M均可刺激細胞增生。故由上述結果，選擇投予10-9M~10-5M之大豆異黃酮素，培養48小時，作為後續實驗的投予濃度及培養時間點。投與10-9M~10-5M之大豆異黃酮素，抽取細胞內的DNA進行測試，發現可顯著增加MC3T3-E1細胞內DNA的量，以10-8M與10-7M兩組所增加細胞內DNA的效果最佳，此符合MTS assay所得到的結果。當異黃酮素之添加濃度為10-9M~10-6M，可顯著刺激MC3T3-E1細胞內鹼性磷酸酶(ALP)的酵素活性，尤以10-7M組刺激MC3T3-E1細胞內ALP的活性最佳。投與大豆異黃酮素，抽取細胞內的蛋白質，以西方墨漬法進行骨蛋白(Osteopontin, OPN、第一型膠原蛋白)表現量的測定，發現投予10-9M~10-7M三組，OPN、第一型膠原蛋白的表現量較高。投予造骨細胞株異黃酮素，抽取細胞內的RNA，給予OPN primers，進行RT-PCR的測試，發現OPN mRNA在各組的表現量並無顯著的差異。故本研究之結論為，大豆異黃酮素可刺激老鼠造骨細胞株之細胞存活率與增生情形，提高DNA的含量與ALP的酵素活性，且可增加MC3T3-E1細胞內骨蛋白(Osteopontin、Type I collagen)的表
    The proliferation effect of isoflavones(genistein and daidzein) were investigated using an MTS assay and DNA content on MC3T3-E1 cells. The samples were tested at levels of 10-9 to 10-5M of isoflavones for 24, 48, 72 and 96 hr incubation. The results obtained from the two assys were consisted and also exhibited dose-dependent effects except 10-5M. At the level of 10-8 and 10-7M, isoflavones had the best result for cell proliferation. Genistein and daidzein stimulated an increase in alkaline phosphatase(ALP) activity on MC3T3-E1 cells in a dose-dependent manner(10-9 ~10-7M). Isoflavones treatment(10-9 ~10-7M) significantly increase the production of osteopontin(OPN) and Type I collagen protein, but not on OPN mRNA expression. In summary: isoflavones at dietary achievable level(10-9 ~10-7M) stimulated the proliferation and DNA content of MC3T3-E1 cells. Isoflavones also increased bone marker synthesis(ALP, OPN and Type I collagen) and could be beneficial for bone growth.
    資料類型: thesis
    顯示於類別:[保健營養學系暨研究所] 博碩士論文

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