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研究生:林靜吟
研究生(外文):Ching-Yin Lin
論文名稱:第十一型介白質和其α-受體在正常及無胚胎懷孕之人類子宮蛻膜及絨毛的表現情形
論文名稱(外文):Expression of IL-11 and IL-11Rα in human decidua and villi of normal and anembryonic pregnancies
指導教授:何弘能何弘能引用關係
指導教授(外文):Hong-Nerng Ho
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:免疫學研究所
學門:醫藥衛生學門
學類:醫學學類
論文種類:學術論文
論文出版年:2001
畢業學年度:89
語文別:中文
論文頁數:78
中文關鍵詞:第十一型介白質子宮蛻膜
外文關鍵詞:IL-11decidua
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第十一型介白質(IL-11) 在鼠類胚胎著床所引起的子宮基質細胞蛻膜化反應過程扮演了一個重要角色。在本篇論文中我們檢測了IL-11和其α受體(IL-11Rα)在下列組織的表現和分佈情形,包括不同月經週期的子宮內膜、早期正常懷孕和無胚胎懷孕 (anembryonic pregnancy)的子宮蛻膜(decidua)和絨毛(villi)。另外,訊息傳遞分子-gp130 mRNA在這些組織的表現情形也同樣被偵測。高表現量的IL-11 蛋白質在早期懷孕的子宮蛻膜和絨毛組織被偵測到,特別是表現在子宮蛻膜基質細胞、腺體上皮細胞、及絨毛組織上的合體滋養細胞(syncytiotrophoblast)和滋養細胞(cytotrophoblast)。我們也利用即時定量聚合酉每 反應(real-time quantitative RT-PCR)來分析IL-11 mRNA的表現量。IL-11 mRNA從分泌期的子宮內膜開始被偵測到,而在正常懷孕的子宮蛻膜和絨毛達到高量表現。然而,anembryonic pregnancy的子宮蛻膜和絨毛的IL-11 mRNA表現量都有明顯的下降。IL-11Rα和gp130在增生期和分泌期的子宮內膜、正常懷孕和無胚胎懷孕的子宮蛻膜和絨毛都有表現,其表現量差異卻不大。IL-11隨月經週期的變化和表現位置都強烈暗示它在人類子宮基質細胞蛻膜化過程扮演了一個重要角色。此外,在無胚胎懷孕中發現IL-11表現量大量減少也顯示IL-11可能對滋養細胞的生長和invasion很重要,或許將來可以把IL-11運用在生殖領域的臨床治療上。

Interleukin-11( IL-11) is crucial in the decidualization of the uterine stroma to prepare for the implantation of blastocyst in the mouse. This study examined the localization and expression of IL-11 and IL-11 receptor α (IL-11Rα) in human endometrium throughout the menstrual cycle and in the decidua and villi of early normal pregnancy and in those of anembryonic pregnancy. The mRNA expression of the signaling component, gp130, in endometrium, decidua, and villi tissue was also examined. Immunoreactive IL-11 was highest in the decidua and villi of early normal pregnancy and present in decidualized stromal cells, glandular epithelial cells, syncytiotrophoblast cells, and cytotrophoblast cells. The IL-11 mRNA expression was verified by real-time quantitative RT-PCR. It was detected in the secretory phase and showed much excess in early normal pregnancy. The decidua and villi of anembryonic pregnancy had a significant decrease in IL-11 mRNA expression. Similar expression pattern and constant amount of IL-11 Rα and gp130 mRNA were detected in endometrium of proliferative phase and secretory phase and in decidua of early pregnancy and anembryonic pregnancy . The cyclical changes in amount and cell type specific expression suggest an important role of IL-11 in the decidualization of stromal cells. The decreased amounts of IL-11 in anembryonic pregnancy also suggests a potential role in the development and invasion of trophoblast cells and may provides another clinical use of IL-11 in reproduction filed.

目錄
論文口試委員審定書………………………………………………..
授權書……………………………………………………………….. ii
電子檔案上網授權書……………………………………………….. iii
誌謝………………………………………………………………….. iv
中文摘要…………………………………………………………….. v
英文摘要…………………………………………………………….. vi
第一章 緒論………………………………………………………… 1
第一節胚胎著床與子宮內變化……………………………… 1
第二節胎兒存活的可能機制………………………………… 3
第三節IL-11的角色…………………………………………... 7
第四節研究動機及目的……………………………………… 10
第二章 實驗材料與方法…………………………………………… 12
第一節實驗材料……………………………………………… 12
2.1.1 實驗材料…………………………………………… 12
2.1.2 反應試劑與溶液…………………………………… 12
第二節實驗方法……………………………………………… 13
2.2.1 細胞株細胞的冷凍與解凍………………………… 13
2.2.2 分離人類周邊血液單核淋巴細胞………………… 14
2.2.3 RNA的萃取、定量、品質確認…………………. 14
2.2.4 反轉錄反應………………………………………… 16
2.2.5 聚合酉每 連鎖反應…………………………………. 16
2.2.6 即時定量聚合酉每 連鎖反應分析………………… 18
2.2.7 石蠟包埋檢體……………………………………… 21
2.2.8 冷凍包埋檢體……………………………………… 22
2.2.9 Cytospin……………………………………………... 23
2.2.10 H&E 染色…………………………………………. 23
2.2.11 免疫染色…………………………………………... 24
2.2.12 構築IL-11Rα 目標基因………………………….. 25
2.2.13 原位雜交法………………………………………... 28
2.2.14 北方墨點轉漬法…………………………………. 31
第三章 結果…………………………………………………………. 34
第一節全RNA的萃取結果…………………………………... 34
第二節以RT-PCR方法偵測IL-11、IL-11Rα、及gp130 mRNA在人類不同組織中的表現…………………………… 34
第三節以real-time quantitative PCR分析IL-11和IL-11Rα mRNA在人類子宮內膜、蛻膜、及絨毛的表現量………… 35
第四節不同時期子宮內膜及normal pregnancy和anembryonic pregnancy病人子宮蛻膜及絨毛之型態變化……….. 37
第五節以免疫化學染色研究IL-11在人類子宮內膜、蛻膜、及絨毛的分佈……………………………………………… 38
第六節以原位雜交法研究IL-11Rα mRNA在人類子宮內膜、蛻膜、及絨毛的分佈情形……………………………… 38
第四章 討論…………………………………………………………. 41
參考文獻……………………………………………………………... 46
圖表與說明…………………………………………………………... 53
附錄…………………………………………………………………... 75

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