臺灣博碩士論文加值系統

老年人容易因為骨質流失而產生骨質疏鬆症。研究發現，停經後的婦女因骨質流失的速度較快，亦隨之而增加罹患骨質疏鬆症的機率，給予雌激素(estrogen，E2)的補充，則可有效減緩骨質流失，但卻有增加罹患子宮癌和乳癌的風險。植物中天然存在的植物性雌激素，有類似於雌激素的結構，並具有與雌激素受體結合的能力。臨床上證據顯示，植物性雌激素，例如異黃酮素，可增加女性的骨量，減緩骨質的流失。但是植物性雌激素與骨質代謝相關的造骨細胞生長、代謝分子標誌等的相關機制並不完全清楚。因此本實驗對異黃酮素如何影響造骨細胞進行分析並與E2的作用途徑作比較。實驗中取用異黃酮素為genistein、daidzein以及biochanin A ( genistein的前趨物)。三種異黃酮素均可促進MC3T3-E1造骨細胞生長，以genistein生長促進效果較佳，故以此作為後續實驗用藥。實驗結果發現，genistein促進造骨細胞增生的效果，可被E2的拮抗劑ICI 182,780所抑制，如同E2一樣，且E2與genistein均可促進和ER相關的collagen以及ALP基因表現，且其反應可被ICI182,780所抑制。MC3T3-E1細胞內具有ERα及ERβ的基因和蛋白質的表現，因此推測genistein可能經由與細胞內ER結合，達到促進細胞增生的效果。以ERK1/2的抑制劑PD98059共同處理MC3T3-E1，可抑制24% E2產生的增生效果，和54% genistein所引起之增生效果。ERK1/2另一抑制劑U0126，則無法抑制由E2以及genistein所引起的增生效果。AKT抑制劑LY294002抑制E2和genistein所促進的增生效果。推測E2與genistein的作用亦可部分透過與膜上受器結合，引起細胞內訊息傳遞以達促進造骨細胞增生。對於細胞週期方面，E2與genistein可促進造骨細胞的分裂，以達到促進增生的效果。因此綜合以上各結果，genistein促進造骨細胞增生的途徑住要是主要是一須ER參與的訊息傳遞途徑。

Osteoporosis associated with estrogen deficiency is the most common cause of age–related bone loss. Current hormone replacement therapy (HRT) can resolve most postmenopausal problems including osteoporosis. However, long term use of HRT has been shown to be associated with elevated risks of breast and endometrial cancers. Phytoestrogen, a group of natural plant-derived compounds, is structurally similar to the estrogen and has estrogen receptor binding ability. Recent clinical evidences showed that phytoestrogens, such as isoflavones, could increase women’s bone mass and desiderate bone loss. However, the mechanism of how phytoestrogen affects bond density is still poorly understood. In this thesis, the effects of isoflavones on bond building osteoblast cells were analyzed. The results showed that all three isoflavone compounds, genistein, daizein, biochanin A used in this study could promote the cell proliferation of MC3T3-E1 osteoblast cells, with genistein exhibited the highest ability. Genistein was also found to increase the cell cycle progression rate. The transition of G1 to S phase of the cell cycle was elevated by genistein. With flow cytometry and RT-PCR, he presence of ERα and ERβ in MC3T3-E1 cells were confirmed. Co-treatment of MC3T3-E1 with estrogen antagonist ICI182,780 abolished most the activated cell proliferation induced by genistein. Administration of ERK1/2 inhibitors PD98059 but not U0126, suppressed the genistein induced cell proliferation to 78%. LY294002, an AKT inhibitor, also blocked genistein promoted cell proliferation to 61%. Moreover, genistein increased the mRNA levels of collagen and ALP in MC3T3-E1 cells. Similar results were obtained with estrogen. In conclusion, the results indicated that the effect of genistein as well as estrogen on osteoblast cell proliferation could be mediated through at least two pathways, mainly through an estrogen receptor dependent pathway and partially via a membrane receptor cellular signaling pathway.

中文摘要…………………………………..…………………..I
英文摘要……………………………………………………..III
壹、緒論………………………………..……………………..1
貳、研究目的………………………………………………..15
參、方法與材料……………………………………………..16
肆、結果……………………………………………………..27
伍、討論……………………………………………………..32
陸、參考文獻………………………………………………..39
柒、圖表……………………………………………………..44
捌、附錄……………………………………………………..65

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