目的：轉化生長因子-（TGF-）在肝纖維化中扮演了重要的角色。本研究探討骨形態發生蛋白-2（BMP-2）調控肝纖維化的作用及機制。方法：我們利用肝纖維化的人體組織微陣列分析(TMA)與接受膽管結紮（BDL）手術或四氯化碳（CCl4）給藥誘導肝纖維化的小鼠模型中觀測BMP-2的表現。我們進一步使用腺病毒介導的BMP-2基因遞送來測試對肝纖維化的預防效果。小鼠的初代肝星狀細胞(pHSC)、細胞株HSC-T6肝星狀細胞和Clone 9肝臟細胞，則用來研究BMP-2和TGF-1之間的相互作用。結果：BMP-2在人類肝纖維化組織和動物肝纖維化模式中的表現顯著降低，並與肝臟TGF-1表現量呈負相關。 BMP-2基因傳遞緩解了動物肝纖維化模式中的血清中肝臟相關指數的升高，HSC活化和肝纖維化的相關標誌物。此外，外加TGF-1可根據劑量依序抑制BMP-2的表現。反之，BMP-2治療顯著抑制TGF-1及其I型和II型訊息傳遞受體的表現達，以及pHSC中誘導的Smad3磷酸化。BMP-2治療除抑制其對細胞增殖和轉移的作用外，BMP-2治療也顯著減弱了因TGF-1刺激所造成的-平滑肌肌動蛋白和纖維蛋白的高度表現，並逆轉小鼠肝臟星狀細胞中因T GF-1刺激所造成的上皮 - 間質轉換。結論：綜合我們觀察的結果，BMP-2明顯的表現出其對於肝纖維化的保護與抑制效果，而BMP-2與TGF-1之間的相互調控可能是BMP-2潛在作為在肝纖維化機轉中的抗纖維化機制。 因此，BMP-2可能可以作為治療肝纖維化的新型治療標的。

Objective: Transforming growth factor-β (TGF-β) plays a central role in hepatic fibrogenesis. This study investigated the function and mechanism of bone morphogenetic protein-2 (BMP-2) in regulation of hepatic fibrogenesis. Methods: BMP-2 expression in fibrotic liver was measured in human tissue microarray and mouse models of liver fibrosis induced by bile duct ligation (BDL) surgery or carbon tetrachloride (CCl4) administration. Adenovirus-mediated BMP-2 gene delivery was used to test the prophylactic effect on liver fibrosis. Primary hepatic stellate cells (HSC) and cell lines, HSC-T6 cells and Clone 9, were used to study the interplay between BMP-2 and TGF-β1. Results: Hepatic BMP-2 expression was significantly decreased in either human fibrotic tissues or mice fibrosis models, with a negative correlation with hepatic TGF-β1 contents. BMP-2 gene delivery alleviated the elevations of serum hepatic enzymes, HSC activation markers, and liver fibrosis in both models. Mechanistically, exogenous TGF-β1 dose-dependently reduced BMP-2 expression, whereas BMP-2 significantly suppressed expression of TGF-β and its cognate type I and II receptor peptides, as well as the induced Smad3 phosphorylation levels in primary mouse HSCs. Aside from its suppressive effects on cell proliferation and migration, BMP-2 treatment prominently attenuated the TGF-β1-stimulated α-smooth muscle actin and fibronectin expression, and reversed the TGF-β1-modulated epithelial-to-mesenchymal transition marker expression in mouse HSCs. Conclusions: The mutual regulation between BMP-2 and TGF-β1 signaling axes may constitute the anti-fibrogenic mechanism of BMP-2 in the pathogenesis of liver fibrosis. BMP-2 may potentially serve as a novel therapeutic target for treatment of liver fibrosis.

書名頁……………..………………………………………………….. i
論文審定書………..……………… ………………………………….. ii
論文公開授權書……………………………………………………… iii
誌謝………………………………………………………………….… iv
中文摘要………………………………………………………….….. v
英文摘要………………………………………..……………………. vi
目錄………………………………………..…………………….……. vii
縮寫表………………………………………..……………………..… viii
圖次………………………………………..……………………..…… ix
論文正文………………………………………………………..……… x
參考文獻……………………………………………………….…… 19
附錄……………………………………………………………………. 52

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