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系統識別號 U0026-1407201116524000
論文名稱(中文) 探討星狀細胞中CEBPD調控ZNF179基因轉錄
論文名稱(英文) Investigation of CEBPD-mediated ZNF179 transcription in astrocytes
校院名稱 成功大學
系所名稱(中) 藥理學研究所
系所名稱(英) Department of Pharmacology
學年度 99
學期 2
出版年 100
研究生(中文) 黃靖芳
研究生(英文) Jing-Fang Huang
學號 s26981034
學位類別 碩士
語文別 中文
論文頁數 64頁
口試委員 指導教授-王育民
口試委員-李宜釗
口試委員-黃阿敏
中文關鍵字 星狀細胞  人類膠質瘤細胞  人類神經瘤細胞  神經發炎  阿茲海默氏症 
英文關鍵字 CEBPD  ZNF179  astrocytes  U373MG  SH-SY5Y  neuroinflammation  HuR  Alzheimer’s disease 
學科別分類
中文摘要 神經發炎 (neuroinflammation) 已被認為在神經病變及退化扮演重要成因之一,但是參與影響阿茲海默症的發炎分子機轉仍有許多未知。CEBPD為CCAAT/enhancer-binding proteins (C/EBP) 家族成員之一。已知CEBPD (human)參與細胞的凋亡、分化及免疫和發炎反應。先前的研究發現,受到澱粉樣蛋白 (amyloid-β peptides) 及促發炎因子的活化,有病斑 (plaque) 形成的神經細胞周遭之星狀細胞 (astrocytes) 能觀察到CEBPD增加情形。Znf179 (mouse)為RING finger 家族成員之一,目前僅知在神經分化過程中扮演重要角色。在本研究中,我們發現Znf179的表現可延緩TNFα所誘發之HeLa細胞凋亡進程,此現象產生一有趣的問題,讓我們想檢測在星形細胞中CEBPD是否有助於 ZNF179 (human)轉錄活化,幫助細胞在炎症環境下生存。實驗發現處理發炎因子IL -1β及TNF可以誘導U373MG細胞中CEBPD和ZNF179的表達。再者藉由報導基因分析結果,我們發現,CEBPD能誘導 ZNF179啟動子活性,反之ZNF179則無法活化CEBPD啟動子。此現象與在小鼠系統中結果是一致,這表明CEBPD是ZNF179上游的基因。此外,利用免疫螢光染色在APP老鼠腦組織切片上可以觀察到澱粉樣蛋白斑附近所聚集的星狀細胞會大量表現CEBPD和Znf179,同時我們證明了星狀細胞中,HuR可能在CEBPD介導 Znf179活化中扮演角色,綜合以上結果,可以解釋炎症反應情況中,星形膠質細胞比神經元更具耐受性。
英文摘要 Neuroinflammation has been known to play an important role in neurodegenerative diseases. However, the inflammation-related molecular mechanisms in pathogenesis of Alzheimer's disease (AD) remain largely unclear. CEBPD, a member of CCAAT/enhancer-binding protein (C/EBP) family, participates in the regulation of various physiological processes including cell apoptosis, differentiation and immune responses. We demonstrated that CEBPD (human) activation in astrocytes is stimulated by A peptides and proinflammatory factors. Preliminary results suggested that Znf179 (mouse), a RING family member, plays a functional role in neuron differentiation. In this study, we found that Znf179 can resistant to TNFα-induced cell death of HeLa cells. It raises an interesting issue for us to test whether CEBPD contributes to ZNF179 (human) transcriptional activation in astrocytes and help the cells to survival in an inflammatory environment. IL-1β and TNF can induce both expressions of CEBPD and ZNF179 in U373MG cell. By using a reporter assay, we found that the CEBPD can induce the ZNF179 reporter activity, not vice versa. This phenomenon is also coincident in mouse system, suggesting that CEBPD is the upstream of ZNF179 gene in general. Moreover Cebpd (mouse) expression is coincident with Znf179 in the activated astrocytes surrounding the A plaques by immunofluroence assay. In addition, we demonstrated that HuR may play an important role in CEBPD-mediated Znf179 activation in astrocytes. Taken together, these results provide an explanation for the reason why astrocytes is more resistant than neuron in an inflammatory situation.
論文目次 中文摘要………………………………………………………..……….. I
Abstract…………………………………………….…………………… II
致謝………………………………………………………….…………… III
目錄………………………………………………………………………. V

第一章 緒論
第一節 前言…………………………………………………….............. 1
第二節 神經及週遭細胞在神經退化性疾病中的功能及特性…….…. 2
第三節 CEBPD之生理功能與特性….………………………………... 3
第四節 Brain Finger Protein ZNF179之生理功能與特性….................. 4
第五節 Hu家族蛋白................................................................................ 5
第六節 研究動機…………………………………………...................... 6
第二章 實驗材料及方法
第一節 來源………………………………………………….................. 8
第二節 實驗方法…………………………………………….................. 13
第三章 實驗結果
第一節 ZNF179延緩細胞凋亡的進程………………………………... 30
第二節 TNF誘導人類膠質瘤細胞中ZNF179蛋白質表達…………. 31
第三節 App轉基因鼠腦組織中Znf179過度表現在活化態星狀細胞 31
第四節 星狀細胞上CEBPD活化ZNF179轉錄及轉譯活性……….... 32
第五節 在U373MG細胞中CEBPD的持續存在會支持ZNF179蛋白穩定表現………………………………………………………. 33
第六節 U373MG細胞中 HuR可以穩定CEBPD mRNA表現............. 33
總結……………………………………………………………………… 35
第四章 討論………………………………………………………………… 36
第五章 參考文獻…………………………………………………………… 41
附圖
Fig. 1. ZNF179 attenuates TNF/MMS-induced death of HeLa cell…… 47
Fig. 2. Expressions of CEBPD with ZNF179 are coincident in TNF- treated cells.……………………………........................................ 49
Fig. 3. Both Cebpd and Znf179 are observed in the activated astrocytes in APP mice………………….…………………………………... 50
Fig. 4. CEBPD is coincident with ZNF179 expression in TNF-induced U373MG cells................................................................................. 51
Fig. 5. CEBPD postively regulates ZNF179 reporter activity in
U373MG cells................................................................................. 52
Fig. 6. ZNF179 correlates with sustained CEBPD expression in
astrocytes.……………………………………………………....... 55
Fig. 7. RNA binding protein HuR could participate in CEBPD-regulated ZNF179 promoter activity in astrocytes.………………………… 56
附錄…………………………………………………………………………… 58
作者簡介………………………………………………………........................ 64
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