臺灣博碩士論文加值系統

B型肝炎病毒的感染會造成急性甚至慢性肝炎的發生，而慢性肝炎的患者具有罹患肝硬化或是肝癌的高度風險。目前雖然已經有B型肝炎疫苗的注射以預防，但對於帶原者卻沒有很好的治療方式，開發有效的治療B型肝炎藥物仍是迫切需要的研究。中藥小柴胡湯在過去常被用於治療肝病，柴胡以及黃芩為其中兩種主要成份，在篩選抑制B型肝炎病毒表面抗原的產生時，發現小柴胡湯中的黃芩具有良好的活性。本論文主要探討黃芩以及其中的一種化合物 Wogonin 抑制B型肝炎病毒的作用機制。在人類肝癌細胞中，黃芩與 Wogonin 除了可以抑制B型肝炎病毒表面抗原的產生，尚可減少B型肝炎病毒複製的中間產物RNA及病毒顆粒。為了瞭解黃芩和Wogonin 對B型肝炎病毒基因表現的作用機制，利用 Luciferase reporter assay 分析B型肝炎病毒主要的四個啟動子在肝癌細胞中的活性，實驗的結果明確看到黃芩和 Wogonin 都會明顯的抑制核心抗原的活性，而當外送核心抗原啟動子到腎臟上皮癌細胞293T 時，藥物對啟動子的抑制現象則消失，此結果表示黃芩和 Wogonin 對於核心抗原啟動子的抑制是具有肝組織特異性的。當外送一些肝組織特有或是肝組織特多的轉錄因子到人類肝癌細胞時，可以看到當PGC-1�悀j量表現時，黃芩對核心抗原啟動子的抑制效果減弱 ; 而當 CEBP/�恁BPPAR�� 大量表現時，Wogonin 對核心抗原啟動子的抑制效果減弱， Wogonin 與黃芩均具有抗B型肝炎病毒的效果，黃芩與 Wogonin 可能是透過不同的機轉抑制B型肝炎病毒的基因表現及複製，黃芩中還有其他有效成份值得進一步探討。

Hepatitis B virus (HBV) infection causes acute and chronic hepatitis, and the affected patients have an increased risk of developing liver cirrhosis and hepatocellular carcinoma (HCC). Although a preventive vaccine for HBV is
available, effective drugs to eradicate HBV in chronic carriers are still urgently needed.The Chinese herbal medicine Xiao-Chai-Hu-Tang consists of Bupleuri radix and Scutellaria radix (HD-1S) and is widely used for treatment of liver disease. In searching of compounds on suppression of hepatitis B surface antigen (HBsAg) production, our laboratory previously have shown that HD-1S strongly suppressed hepatitis B surface antigen (HBsAg) in human hepatoma cells. In this study, I demonstrated that the aqueous extract of HD-1S and one of its components Wogonin had potent anti-HBV activity. Both HD-1S and Wogonin not only suppressed HBsAg production but also decreased HBV transcripts and HBV viral particles in cultured human hepatoma cells. To understand the molecular mechanism of suppression of HBV gene expression by HD-1S and Wogonin, the effects of HD-1S and Wogonin on four viral promoter activities using luciferase as a reporter were examined. This study showed clearly that both HD-1S and Wogonin selectively suppresses core promoter (CP) of HBV. Such suppressive effects of HD-1S and Wogonin are liver-specific because no suppressive activity of HD-1S and Wogonin was observed when CP activity was assayed in non-liver 293T cells. Furthermore, ectopic expression of PGC-1�� and PPARr/CEBP�� abolished the suppressive effect of HD-1S and Wogonin on HBV CP activity, respectively. Therefore, HD-1S and Wogonin may selectively modulate transcriptional machinery of human liver cells to suppress HBV gene expression and replication. Results from this study also suggested that some components other than Wogonin might also have novel anti-HBV activity and deserve futher
investigation in the future.

中文摘要…………………………………………………………………1
英文摘要…………………………………………………………………2~3
緒論………………………………………………………………………4~12
材料與方法………………………………………………………………13~29
結果………………………………………………………………………30~39
討論………………………………………………………………………40~44
參考文獻…………………………………………………………………45~50
圖表………………………………………………………………………51~79

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