2011-08-012024-05-13https://scholars.lib.ntu.edu.tw/handle/123456789/649602摘要：胚胎著床機制是生殖醫學面臨的重大問題，胚胎著床的重要因素包括良好的受精卵與健康的著床環境，也就是子宮內膜的環境。現有的醫療技術對於受精卵品質的調控已經發展到達一個程度，如何調控子宮內膜的最佳化成為生殖醫學最大的挑戰。本團隊近年來致力於不孕症生殖醫學的相關研究，在卵巢過度刺激症的研究著重於血管通透性與抗細胞凋亡兩大機制，三年內共有五篇發表於JCEM、 Human Reproduction、Endocrine 期刊之論文，在研究過程中，我們發現HCG 會刺激黃體的抗凋亡。近年來有研究顯示 HCG 可使用於胚胎植入的過程中，我們的初步研究成果顯示HCG 會誘發子宮內膜細胞產生抗凋亡分子Mcl-1，因此我們推論Mcl-1 在子宮內膜應該具有重要的角色，而且可能與胚胎著床機制有關。因此在本三年期計畫中我們主要將探討Mcl-1在子宮內膜細胞的調控機制與其對胚胎著床機制的影響。本計畫的分年目標與執行策略分別為第一年， 以動物模式探討自然月經週期情況下Mcl-1 在子宮內膜表現的情況，主要使用的方法為免疫組織染色，我們也將利用人類子宮內膜細胞RL95-2 探討生殖內分泌相關荷爾蒙對Mcl-1 產生的效應，並運用化學抑制劑探討個別的調控分子機制。第二年， 主要將探討Mcl-1 對胚胎著床的影響，主要的研究策略與方法是利用RNA 干擾技術抑制Mcl-1 在子宮內膜細胞的表現，再以體外胚胎著床測試方法驗證Mcl-1 表現對著床的效應，分子機制的探討將以免疫螢光染色及流式細胞儀分析Mcl-1 是否影響內膜細胞膜表面固著分子的表現為主軸。第三年， 主要將探討Mcl-1 在活體內子宮內膜細胞的表現對胚胎著床的效應，我們已經著手在建立子宮內膜細胞專一性Mcl-1 抑制基因轉殖鼠，主要運用galectin-9 promoter 調控細胞專一性，並以tetracyclin 調控Mcl-1 shRNA 表現，我們將以此模式配合胚胎植入技術探討Mcl-1 的表現對內膜健康情況及著床的效應。我們也將嘗試以基因治療方式探討Mcl-1 表現對內膜細胞健康情況及著床的效應。本研究成果將能證明抗凋亡分子 Mcl-1 對子宮內膜細胞的重要性，也提供生殖相關荷爾蒙對Mcl-1 的調控機制，Mcl-1 的基因治療也將對人類生殖醫學在子宮內膜環境的改造上有重大的影響。<br> Abstract: The implantation of the embryo into the maternal uterus is a critical step in humanreproduction. Myeloid cell leukemia-1 (Mcl-1), a member of Bcl-2 family, can inhibitapoptosis by inhibiting Ca2+ signals within mitochondrial. Apoptosis is believed to playcritical roles in endometrium. However, the regulation and the role of Mcl-1 in endometriumand its effect on embryo implantation are unclear. The specific aim of this project is toinvestigate the roles of Mcl-1 in embryo implantation. The research designs of the threeyears project are as follows. In the first year, the aim is to evaluate the expression of Mcl-1in endometrial tissue in natural cycle in mouse model by immunohistal chemical stainingand to clarify the effect of pregnancy related endocrine molecular estradiol, and progesteroneon the regulation of Mcl-1 in human endometrial cells with RL-95-2 cell line, furthermore,molecular biology methodologies will be used to clarify the signaling transduction pathwaysand transcriptional regulation. In the second year, the aim is to clarify the anti-apoptosisrole and the effect of Mcl-1 on embryo implantation, Mcl-1 is a calcium regulatory protein,and the anti-apoptosis role of Mcl-1 will be verified by series apoptosis assays, the effect ofMcl-1 on adhesion molecules of endometrial epithelial cells will be verified by florescencestaining based flow cytometry analysis. In the third year, the aim is to verify the role ofMcl-1 on endometrial cells and its effect on embryo adhesion in vivo. We will establish anendometrial cells specific and Mcl-1 conditional know-down mouse strain by usinggalectin-9 gene promoter- Tetracycline-controlled Mcl-1 shRNA transcriptional activationstrategy. The transgenic mouse will be used to evaluate the effect of Mcl-1 on endometriumin vivo by histopathology judgment; furthermore, embryo implantation technology will beused to evaluate the effect of Mcl-1 expression on embryo implantation and the birth rate offetus. Adenovirus-mediated Mcl-1 gene-therapy will be used to test the recover effect in theMcl-1 knock-down mice and evaluate the function by histopathology judgment and embryoimplantation technology. Results obtained from this project may provide importantinformation for the clinically relevant role in enhancing embryo implantation.胚胎著床子宮內膜Myeloidcellleukemia-1embryo implantation endometrium Myeloid cell leukemia-1