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研究生:林元鈺
研究生(外文):Yuan-Yu Lin
論文名稱:探討Klf6在Mir122a缺失時所引起的肝臟纖維化中所扮演的角色
論文名稱(外文):Contribution of Klf6 in liver fibrosis in Mir122a deficiency
指導教授:鄒安平
指導教授(外文):Ann-Ping Tsou
學位類別:碩士
校院名稱:國立陽明大學
系所名稱:醫學生物技術暨檢驗學系
學門:醫藥衛生學門
學類:醫學技術及檢驗學類
論文種類:學術論文
論文出版年:2012
畢業學年度:100
語文別:中文
論文頁數:51
中文關鍵詞:肝臟纖維化肝臟星狀細胞
外文關鍵詞:Klf6Liver fibrosismiR-122aHepatic Stellate Cell
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Mir122a是一個可以抑制腫瘤形成的微小核醣核酸,由我們實驗室近年來的研究指出,在小鼠當中,Mir122a的缺失會引發漸進式的肝癌病程,而肝臟纖維化也包含在此病程之中。肝臟纖維化的形成,通常是由不斷的發炎反應所引起的,因為發炎反應的關係,一些發炎反應的因子會被釋放出來,像是Tgf-1以及Il-6,然而這些因子,會刺激Hepatic stellate cell (HSC)的活化,進一步造成Collagen I的累積與一些在肝臟纖維化這個主題常用到的指標:matrix metalloproteinase (MMP), tissue inhibitor of metalloproteinase (TIMP)表現量上升。
Krüppel-like factor 6 (Klf6)是Krüppel -like factor家族當中其中一種轉錄因子,它可以在轉錄層次進行正向調控或是負向調控。在近年來的研究當中也指出,在活化的HSC以及纖維化的肝臟當中,Klf6的表現量會上升,而Klf6的轉錄目標基因也包含與肝臟纖維化息息相關的指標,像是Col1a1、Tgfb1、Serpinh1。至今,我們仍不了解Klf6是如何調控肝臟纖維化發生的。
在本論文當中,我將研究(1)在Mir122a缺失的條件下,Klf6、Klf6 target genes以及fibrosis associated genes的表現量與正常老鼠有何不同。(2)探討Klf6調控肝臟纖維化的分子機轉。
由實驗結果得知,Klf6、Klf6 target genes以及fibrosis associated genes的表現量在Mir122+/+會比Mir122a-/-的表現量來的高,而Tgf-1的分泌量也具有相同的情形,藉由Sirius red實驗結果也得知Mir122a-/-產生的肝臟纖維化會隨著年齡增加而嚴重。於是我更進一步的使用hydrodynamic injection將shKlf6由尾靜脈打入小鼠肝臟內,發現shKlf6能有有效抑制Klf6、Klf6 target genes以及fibrosis associated genes在Mir122a-/-的基因表現量,肝臟纖維化的collagen堆積還有HSC的活化。並且藉由實驗數據推論出一套Klf6調控肝臟纖維化的分子機轉。這也證實了Klf6在調控肝臟纖維化的課題中,的確扮演著一個相當重要的角色。

MiR-122a is a tumor suppressor microRNA. Mice lacking miR-122a (Mir122-/-) are viable but develop temporally controlled steatohepatitis, fibrosis and hepatocellular carcinoma (HCC). How miR-122a regulates liver fibrosis is unclear. Liver fibrosis is a disease stimulated by multiple inflammatory mediators, such as Tgf-β1 and Il-6, which led to hepatic stellate cell (HSC) activation, collagen accumulation and up-regulation of matrix metalloproteinase (MMP) as well as tissue inhibitor of metalloproteinase (TIMP). Recently, we identified Klf6, a fibrogenic transcription factor as a novel miR-122a target gene. Krüppel-like factor 6 (Klf6) is reported to be associated with HSC activation and elevated level of Klf6 was detected in the fibrosis livers of both human and rodents. To characterize the roles of Klf6 in liver fibrosis of Mir122-/- mice, we investigated (i) the expression of Klf6 target genes and fibrosis associated genes in liver fibrosis under mir-122 deficiency. (ii) the molecular mechanism of Klf6-regulated liver fibrosis.
In Mir122a-/- livers, the expression levels of Klf6, Klf6 target genes and fibrosis associated genes as well as serum level of Tgf-β1 are much higher than that of the wild-type mice. In vivo silencing of Klf6 by the hydrodynamic injection effectively reduced the severity of liver fibrosis, the elevated serum levels of Tgf-β1 and the hallmarks of HSC activation. I integrated the experimental data to infer a new molecular mechanism of Klf6-regulated liver fibrosis. These results also confirm that Klf6 plays a very important role in liver fibrosis.

中英文名詞縮寫對照表…………………………………………………1
中文摘要 ………………………………………………………………2
英文摘要 ………………………………………………………………3
緒論 ……………………………………………………………………4
論文研究動機 …………………………………………………………9
實驗材料 ………………………………………………………………10
實驗方法 ………………………………………………………………12
實驗結果 ………………………………………………………………20
討論 ……………………………………………………………………25
總結 ……………………………………………………………………29
參考文獻 ………………………………………………………………30
圖表 ……………………………………………………………………34
附錄 ……………………………………………………………………49

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