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完整後設資料紀錄
DC 欄位 | 值 | 語言 |
---|---|---|
dc.contributor.advisor | 朱善德(Sin-Tak Chu) | |
dc.contributor.author | Che-Yu Lin | en |
dc.contributor.author | 林哲宇 | zh_TW |
dc.date.accessioned | 2021-06-08T04:45:29Z | - |
dc.date.copyright | 2009-08-11 | |
dc.date.issued | 2009 | |
dc.date.submitted | 2009-07-31 | |
dc.identifier.citation | 1. Bratt, T., Lipocalins and cancer. Biochim Biophys Acta, 2000. 1482(1-2): p. 318-26.
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dc.identifier.uri | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/23169 | - |
dc.description.abstract | Lcn2蛋白屬於疏水性分子結合蛋白家族中的一員,為急性期反應蛋白之一,實驗室先前研究指出小鼠子宮Lcn2蛋白可以透過小鼠動情週期之類固醇荷爾蒙調控,以Lcn2蛋白處理人類子宮內膜細胞株(RL95-2)會刺激IL-8和MCP-1的表現,同時使細胞凋亡停止。子宮內膜異位症是由雌激素刺激子宮內膜細胞過度增生所發生的疾病,而IL-8和MCP-1會參與誘導子宮內膜異位症的形成。為了瞭解Lcn2是否於子宮內膜異位症形成過程中可能參與的生理功能,利用手術建立小鼠子宮內膜異位症的模式以及純化自小鼠子宮分泌液中的Lcn2蛋白對小鼠巨噬細胞進行研究,以瞭解Lcn2蛋白在子宮內膜異位症中經由巨噬細胞所參與的可能分子機制。實驗中發現子宮內膜異位症的小鼠腹腔液中Lcn2濃度較高。此外實驗證實Lcn2於無血清狀態測定巨噬細胞活化因子iNOS、TNF-α均顯示巨噬細胞是Lcn2刺激而活化,同時會使活化的巨噬細胞產生IL-6、GCSF、RANTES、COX-2、MIP-2、MCP-1等與子宮內膜異位症形成相關的因子。推測高濃度的Lcn2在腹腔液中可活化巨噬細胞與誘導子宮內膜異位症的形成可能有密切的關係。 | zh_TW |
dc.description.abstract | Lcn2 protein, a lipocalin, is one of the acute phase proteins. According to our previous data, ovarian steroids regulate the expression of Lcn2 during the estrous cycle. Besides, Lcn2 protein has been found to induce IL-8 and MCP-1 expression in human endometrial carcinoma cell line (RL95-2). There are substantial evidences showing that the progression of endometriosis is estrogen-dependent and that increased levels of cytokines such as IL-8 and MCP-1 are associated with endometriosis formation. In this study, a mouse model of surgically induced endometriosis and a mouse macrophage cell line (RAW264.7) were used to investigate the role of Lcn2 during the development of endometriosis. We demonstrated that during endometriosis formation, the levels of Lcn2 were increased in the peritoneal fluid. Treatment of RAW264.7 cells with Lcn2 resulted in the activation of macrophages. This was accompanied by the elevation of iNOS and TNF-α and the secretion of cytokines, including IL-6, GCSF, RANTES, COX-2, MIP-2 and MCP-1, all of which are factors that facilitate the development of endometriosis. In conclusion, Lcn2 may trigger the activation of macrophages in the peritoneal fluid and thus promote the development of endometriosis. | en |
dc.description.provenance | Made available in DSpace on 2021-06-08T04:45:29Z (GMT). No. of bitstreams: 1 ntu-98-R95b46017-1.pdf: 1382854 bytes, checksum: 2e6941adaab929fa4c75beeb319ea0aa (MD5) Previous issue date: 2009 | en |
dc.description.tableofcontents | 摘要……………………………………….……………………………………………. i
Abstract…………………………………….…………………………………………... ii 縮寫表…………………………………………………………………………………. iii 目錄……………………………………………………………………………………. vi 圖表目錄……………………………………………………………………………... viii 前言……………………………………….……………………………………………. 1 材料與方法……………………………….…………………………………………... 8 小鼠動物實驗………………………………………………………………...…. 8 小鼠子宮液誘發與Lcn2蛋白純化………………………………………... 8 子宮液誘發與收集………………………………………………….... 8 分子篩膠體管柱層析………………………………………………… 8 蛋白質濃縮…………………………………………………………… 9 逆向高效能液相層析…………………………………….…………... 9 質譜分析(膠體內水解) ………………………………………………. 10 小鼠子宮內膜異位系統建立手術………………………………….……... 11 卵巢摘除手術………………………………………………………… 11 血液、腹腔液、腹腔內免疫細胞等樣品收集………………………… 12 小鼠初代巨噬細胞培養……………………………………………………… 13 小鼠巨噬細胞株培養…………………………………………………………… 13 細胞株RNA萃取………………………………………………………………... 14 反轉錄反應……………………………………………………………………… 16 聚合酵素連鎖反應……………………………………………………………… 16 洋菜膠電泳……………………………………………………………………… 17 即時定量聚合酵素連鎖反應…………………………………………………… 18 一氧化氮測量…………………………………………………………………… 18 細胞蛋白質萃取………………………………………………………………… 19 小鼠細胞激素抗體微陣列分析………………………………………………… 19 西方墨點………………………………………………………………………… 20 結果…………………………………………………………………………………… 22 移植子宮組織至腹腔後腹腔液中Lcn2濃度隨時間的變化….…..…………… 22 子宮內膜異位小鼠腹腔液中Lcn2濃度變化…………...……………………… 22 子宮內膜異位腹腔液Lcn2濃度與巨噬細胞關係……...……………………… 23 Lcn2蛋白與RAW264.7巨噬細胞株活化……………………………………… 24 Lcn2蛋白對於RAW264.7巨噬細胞株細胞激素及相關因子的影響………… 25 RAW264.7細胞株Lcn2受體的表現…………………….………………….….. 26 Lcn2與巨噬細胞中MAPK、CREB訊息傳遞的關係………………………...… 27 討論…………………………………………………………………………………… 28 圖表…………………………………………………………………………………… 32 參考文獻……………………………………………………………………………… 53 附錄…………………………………………………………………………………… 64 附錄一 RT-PCR primer 序列……………………………………………….… 64 附錄二 小鼠子宮內膜異位移植組織………………………………………….. 65 圖表目錄 表一、子宮內膜異位移植手術不同天數情況………………………………….. 32 表二、子宮內膜異位移植3週後腹腔液Lcn2濃度及巨噬細胞數目………….. 33 圖一、子宮內膜異位移植手術腹腔液Lcn2變化……………………………..... 34 圖二、子宮內膜異位腹腔液Lcn2濃度與巨噬細胞關係………………….…… 36 圖三、子宮內膜異位腹腔液巨噬細胞一氧化氮濃度測量…………………..… 37 圖四、Lcn2蛋白身份鑑定……………………………………………………..… 38 圖五、Lcn2對於RAW264.7細胞株形態影響………………………………….. 40 圖六、Lcn2對於RAW264.7細胞株活性測量………………………………….. 42 圖七、分析Lcn2對於RAW264.7細胞激素濃度的改變………..……………… 44 圖八、Lcn2對於RAW264.7細胞株IL-6、GCSF、COX2 mRNA表現量的影響 …………………………………………………………………………… 46 圖九、Lcn2對於巨噬細胞與子宮內膜異位相關細胞激素的影響…………...... 48 圖十、Lcn2對於巨噬細胞中Lcn2與Lcn2受體:24p3R、megalin mRNA表現 量的影響………………………………………………………………… 50 圖十一、Lcn2對於巨噬細胞MAPK與CREB活性的影響…………………….. 51 | |
dc.language.iso | zh-TW | |
dc.title | Lcn2蛋白活化巨噬細胞與子宮內膜異位症之相關研究 | zh_TW |
dc.title | Study of the relationship between Lcn2 protein-activated macrophage and endometriosis | en |
dc.type | Thesis | |
dc.date.schoolyear | 97-2 | |
dc.description.degree | 碩士 | |
dc.contributor.oralexamcommittee | 余榮熾(Lung-Chih Yu),張?仁(Ching-Jin Chang),蔡偉博(Wei-Bor Tsai) | |
dc.subject.keyword | 巨噬細胞,疏水性結合蛋白,子宮內膜異位, | zh_TW |
dc.subject.keyword | Lcn2,macrophage,endometriosis, | en |
dc.relation.page | 65 | |
dc.rights.note | 未授權 | |
dc.date.accepted | 2009-07-31 | |
dc.contributor.author-college | 生命科學院 | zh_TW |
dc.contributor.author-dept | 生化科學研究所 | zh_TW |
顯示於系所單位: | 生化科學研究所 |
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