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研究生:潘乃蓁
研究生(外文):Nai-Chen Pan
論文名稱:純植物化合物Cytopiloyne 透過GATA-3 途徑且影響PKCα之路徑抑制脂肪細胞的分化
論文名稱(外文):Cytopiloyne regulated adipogenesis by maintaining GATA-3 expression and affecting the PKCα pathway
指導教授:張力天
指導教授(外文):Lee-Tian Chang
學位類別:碩士
校院名稱:國立中興大學
系所名稱:獸醫學系暨研究所
學門:獸醫學門
學類:獸醫學類
論文種類:學術論文
論文出版年:2010
畢業學年度:98
語文別:中文
論文頁數:51
中文關鍵詞:大花咸豐草萃取物脂肪細胞分化
外文關鍵詞:Cytopiloyneadipogenesis
相關次數:
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有些天然植物化合物具有抑制脂肪細胞分化及誘導脂肪細胞凋亡的能力。脂肪細胞分化包含許多基因表現因子的調控作用,如peroxisome proliferator activated receptor-γ (PPARγ) 及GATA binding protein-3 (GATA-3)。在我們的研究中證實由大花咸豐草萃取出的植物化合物cytopiloyne (CP),可以抑制不同動物來源的脂肪細胞分化。我們也證實出CP透過GATA-3途徑抑制脂肪細胞分化。我們實驗室已有的數據指出CP能調節RAW264.7巨噬細胞及RIN-m5F胰島細胞的PKCα膜到質轉位作用。近年來的文獻亦指出,PKCα和鈣離子之結合蛋白calreticulin (CRT) 的表現情形會影響脂肪細胞分化情形。因此,我們未來也將探討CP對PKCα、CRT 及 Ca2+ 之間的關係。

Certain nature compounds have ability to inhibit adipogenesis and induce apoptosis in adipocytes. Adipocyte differentiation involves a temporally regulation set of gene-expression events, such as peroxisome proliferator activated receptor-γ (PPARγ) and GATA binding protein-3 (GATA-3). In our investigation, we found cytopiloyne (CP), a phytocompound from Bidens pilosa, can reduce the differentiation of adipocytes from various animal sources. We also proved CP inhibits adipocyte differentiation through the GATA-3 pathway in 3T3-L1 cells. Our previous results showed CP can regulate the expression of PKCα translocation in RAW264.7 macrophages and RIN-m5F β-cells. Recent papers indicated that the expression of PKCα and calreticulin (CRT), the major Ca2+-binding protein, can regulate the adipogenesis in 3T3-L1 cells. Therefore, we will focus the adipogenic effect of CP on interaction among PKCα, CRT and Ca2+.

中文摘要 .i

英文摘要 .ii

目次 .iii

第一章 前言 .1

第二章 文獻探討 .2

第一節 脂肪細胞與疾病之相關性 .2

第二節 脂肪細胞之代謝機制 2

一、脂肪細胞之生理功能 .2

二、脂肪細胞研究類型之分類 .3

第三節 天然化合物及脂肪細胞之相關性 .3

第四節 脂肪細胞之分化機制 .4

一、GATA binding protein (GATA) .4

二、Peroxisome proliferator activated receptor (PPARs) .5

三、 CCAAT/enhancer binding proteins (C/EBPs) .6

第五節 脂肪細胞之鈣離子調控機制 .6

一、Protein kinase C .6

(一) PKC 之分類 .6

(二) PKC之結構 .7

(三) PKC之活化機制 .7

(四) PKC和脂肪細胞分化之相關性 .8

二、Calreticulin (CRT) .8

第三章 材料與方法 .10

第一節 細胞培養 (cell culture) .10

第二節 油紅染色法 (Oil Red O staining) .12

第三節 細胞凋亡試驗 (cell viability and apoptosis assays) .13

第四節 小干擾RNA試驗 (siRNA) .13

第五節 RNA萃取及反轉錄聚合酶鏈鎖反應 (RT-PCR) .14

第六節 及時聚合酶鏈鎖反應 (Real-time) .15

第七節 蛋白質萃取及西方墨點法 (Western blotting) 15

第八節 薄膜色層分析法 (Thin Layer Chromatography) .17

第四章 結果 .18

第五章 討論 .29

Reference .33

附圖 .40

附錄 .49


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