慢性肝損傷造成肝臟纖維化。肝臟星狀細胞觸發的發炎產生第一型膠原蛋白過度 的累積最終導致肝硬化,是台灣主要的公共健康問題。在人類肝硬化的樣本中大 麻受體第 1 型(CB1)受到高度的誘導。以四氯化碳(CCl4)誘發 4 週後的大麻受 體第 1 型轉基因培育的小鼠(CB1 overexpression)具有比野生型小鼠較高的肝臟 纖維化的水平。以 Masson’s trichrome stain 和 Immunohistochemistry 染色,顯現 出大麻受體第 1 型轉基因培育的小鼠其肝臟組織中的第一型膠原蛋白比野生型小 鼠的更高。大麻受體第 1 型轉基因培育的小鼠其肝臟組織中大麻受體第 1 型和α- 平滑肌肌動蛋白(肝臟纖維化的標誌物)在免疫組織化學染色的表達均比野生型 小鼠有顯著的增加。CB1 overexpression 會助長肝臟纖維化的進展。這顯示,大麻 受體第 1 型的反義寡核苷酸可能對於肝臟纖維化具有治療的效果。 以四氯化碳誘導肝臟纖維化的 B6 小鼠 隨機接受了 4 週的 CB1 antisense oligonucleotide,CB1 sense oligonucleotide,或大麻受體第 1 型受體阻抗劑 AM251。 B6 小鼠罹患肝臟纖維化之後接受 28 天的 CB1 antisense oligonucleotide 的治療, CB1 antisense oligonucleotide 治療組的肝臟組織其第一型膠原的表達回復到類似正 常小鼠。CB1 antisense oligonucleotide 治療組的肝臟組織中 CB1 受體和α-平滑肌肌 動蛋白的表達也比肝臟纖維化的小鼠顯著降低。在 CB1 antisense oligonucleotide 治 療組,其肝臟 IL-6 的表達較肝臟纖維化組顯著下降。在血清白蛋白的表達方面, CB1 antisense oligonucleotide 治療組較肝臟纖維化小鼠和正常小鼠兩者都高。大麻 受體第 1 型反義寡核苷酸治療組的肝功能得以恢復到正常。 透過大麻受體第 1 型 反義寡核苷酸和 AM251 對大麻受體第 1 型在基因和藥物上的阻斷,不僅削弱了肝臟纖維化的水平並可將肝臟纖維化的小鼠之肝功能修復正常。調控大麻受體第 1 型的信號對肝臟纖維化有治療的潛力。

Liver fibrosis results from chronic injury to the liver and inflammation activation of hepatic stellate cells produce excessive accumulation of type I collagen proteins, ultimately leads to cirrhosis, a major public health in Taiwan. Cannabinoid receptor 1 (CB1) was highly induced in human cirrhotic samples. CB1 transgenic mice (CB1 overexpression) induced by carbon tetrachloride (CCl4 ) for 4 weeks with higher liver fibrosis level than the wild type mice. Collagen type I in liver tissue of CB1 transgenic mice is higher than wild type mice by Masson’s trichrome stain and immunohistochemistry(IHC) stain. Both CB1 and α-SMA Immunohistochemistry
(liver fibrosis biomarker) expression in liver tissue of CB1 transgenic mice significantly increased compared with the wild type mice. Overexpression of CB1 promotes progression of fibrosis. This suggests that CB1 antisense oligonucleotide may have therapeutics effect for liver fibrosis.
B6 mice with CCl4-induced liver fibrosis were randomized to receive CB1 antisense oligonucleotide, CB1 sense oligonucleotide, or the CB1 antagonist AM251 for 4 weeks. After B6 mice with liver fibrosis had 28 days CB1 antisense oligonucleotide treatment,
type I collagen expression in liver tissue of CB1 antisense oligonucleotide therapy group returned to similar with normal mice. The CB1, α-SMA expression in liver tissue of CB1 antisense oligonucleotide therapy group were also significantly decreased compared with the liver fibrosis mice. The hepatic IL-6 expression in CB1 antisense oligonucleotide therapy group was significantly decreased than liver fibrosis group. For serum albumin expression respect, CB1 antisense oligonucleotide therapy group were higher than both liver fibrosis mice and normal mice. The CB1 antisense oligonucleotide therapy group liver function is restored to normal. Genetic and pharmacologic blockade of CB1 by CB1 antisense oligonucleotide and AM251 attenuated liver fibrosis level and can repair liver fibrosis mice hepatic function to normal. Modulation of CB1 signaling has therapeutic potential for liver fibrosis.

Content
I.中文摘要.........................................................................ii II.Abstract..........................................................................iv III.Abbreviation...................................................................vi
IV .Introductions...................................................................1
V.Material and Methods.........................................................7
VI.Results
Overexpression CB1 mice deteriorating liver fibrosis than normal type mice.............................12
CB1 Antisense reduce the AST and ALT level in serum of B6 mice and recover the function of albumin................................................................................................................15
CB1 Expression in Chronic Liver Damage after Treatment of CB1 Antisense.............................19
CB1 Antisense can retard the accumulation of collagen in the liver of Mice with Fibrosis...............22
CB1 Antisense inhibited the activation of hepatic stellate cells..............................................25
CB1 Antisense treatment improved hepatic inflammation response.........................................28
CB1 Antisense promoted the regeneration of damaged liver cells...........................................32
VII.Disscusion.....................................................................34 VIII.Reference.....................................................................36
IX.Attachment.....................................................................40

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