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研究生:李俊彥
研究生(外文):Lee Chun-Yen
論文名稱:小神經膠細胞CD200受器的特性
論文名稱(外文):Characterization of CD200 receptors on microglia
指導教授:吳慶祥
指導教授(外文):Wu Ching-Hsiang
學位類別:碩士
校院名稱:國防醫學院
系所名稱:生物及解剖學研究所
學門:生命科學學門
學類:生物訊息學類
論文種類:學術論文
論文出版年:2007
畢業學年度:95
語文別:中文
中文關鍵詞:小神經膠細胞
外文關鍵詞:microglia
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中文摘要

小神經膠細胞是中樞神經系統中最小的神經膠細胞,它的起源來自於單核球,並有許多細長、具分枝狀的突起。研究指出小神經膠細胞能對其它的神經性細胞造成許多的影響,而這樣的影響被認為是透過這些細胞上的CD200和小神經膠細胞的CD200R進行交互作用後,來調控小神經膠細胞的活性。雖然CD200和CD200R同屬於黏著分子中的免疫球蛋白超級家族成員,不過只有CD200 R才能夠被磷酸化而擁有訊息傳導的功能。本篇論文研究的目的即在探討小神經膠細胞是否有CD200R的存在及其特性。實驗結果顯示,在體外培養的小神經膠細胞的確有CD200R的存在;而在加入LPS的刺激後,小神經膠細胞CD200R的表現有些微增加的情況產生。之前的研究指出,加入抗CD200R的抗體去治療由膠原纖維所引發關節炎的老鼠,可使其症狀得到明顯的改善。因此我們想去觀察加入抗CD200R的抗體對於經由LPS活化過後的小神經膠細胞有何影響。實驗結果顯示經由LPS活化過後的小神經膠細胞,不管是同時加入還是事先加入抗CD200R抗體的處理,小神經膠細胞一氧化氮的累積量都有明顯的增加。
Abstract

Microglia is the smallest glia cell in the central nervous system, which showed thin processes with much branch and originated from circulating monocytes. The researches pointed out that microglia could cause much influence on other neural cells. The latter, in contrast, had also been proposed to modify microglial activation through the interaction of CD200 of cells with microglial CD200R. Although CD200 and CD200R are adhesion molecules belonging to the member of immunoglobulin superfamily, only CD200R can be phosphorylated and has ability to invoke signal transduction. The purpose of the present study is to characterize microglial CD200R if existed. In our data, we found that CD200R did exist on cultured microglia and could be up-regulated after LPS challenge. In the previous study, administration of anti-CD200R antibody could ameliorate collagen-induced arthritis. Therefore, the effects of anti-CD200R antibody on LPS-induced microglial activation were examined. Our results however showed an enhancement of LPS-induced NO accumulation in isolated microglia regardless pre-treated and co-treated with anti-CD200R antibody.
目錄
頁次
目錄 ---------------------------------------------------------------------------------------I
圖目錄 ----------------------------------------------------------------------------------III
中文摘要 -------------------------------------------------------------------------------IV
英文摘要 --------------------------------------------------------------------------------V
第一章 緒言 ----------------------------------------------------------------------------1
第一節 小神經膠細胞 --------------------------------------------------------1
第二節 CD200和CD200R ---------------------------------------------------7
第二章 實驗目的 --------------------------------------------------------------------11
第三章 材料與方法 -----------------------------------------------------------------12
第一節 細胞培養 --------------------------------------------------------------12
第二節 免疫細胞化學染色 --------------------------------------------------14
第三節 免疫螢光雙重染色 --------------------------------------------------14
第四節 亞硝酸根濃度的測定 -----------------------------------------------16
第五節 西方墨點法 -----------------------------------------------------------18
第六節 統計分析 --------------------------------------------------------------20
第四章 實驗結果 -------------------------------------------------------------------21
第一節 小神經膠細胞CD200R的免疫細胞化學染色 ----------------21
第二節 小神經膠細胞與CD200R的免疫螢光雙重染色 -------------21
第三節 LPS活化小神經膠細胞後CD200R的表現 --------------------22
第四節 LPS誘導小神經膠細胞CD200R的表現量增加 --------------22
第五節 抗CD200R抗體對經由LPS活化的小神經膠細胞活性造成的影響 -------------------------------------------------------------------23
第六節 LPS活化小神經膠細胞後MHC II的表現 --------------------24
第五章 討論 -------------------------------------------------------------------------25
第一節 小神經膠細胞的型態變化 ----------------------------------------25
第二節 加入LPS刺激後小神經膠細胞CD200R的變化 ------------26
第三節 加入LPS刺激後給予抗CD200R抗體對小神經膠細胞活性所造成的影響 ----------------------------------------------------------27
第六章 結論 -------------------------------------------------------------------------30
第七章 參考文獻 -------------------------------------------------------------------37

圖目錄
頁次
圖一、混合神經膠細胞培養以及富含小神經膠細胞培養中CD200R的免疫反應 ----------------------------------------------------------------------------31
圖二、混合神經膠細胞培養以及富含小神經膠細胞培養中CD200R的免疫螢光雙重染色 ----------------------------------------------------------------32
圖三、加入LPS刺激後小神經膠細胞CD200R的表現 ---------------------33
圖四、加入不同濃度的LPS刺激後小神經膠細胞CD200R的表現 ------34
圖五、加入LPS與抗CD200R抗體處理後小神經膠細胞一氧化氮的累積量 -------------------------------------------------------------------------------------35
圖六、加入LPS刺激後小神經膠細胞MHC II抗原的免疫螢光染色 ----36
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