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博碩士論文 etd-0729116-161442 詳細資訊
Title page for etd-0729116-161442
論文名稱
Title
正常懷孕與子癲前症孕婦胎盤中的基因差異表現
Differential gene expression in human placentas from preeclampsia and normal pregnancies
系所名稱
Department
畢業學年期
Year, semester
語文別
Language
學位類別
Degree
頁數
Number of pages
47
研究生
Author
指導教授
Advisor
召集委員
Convenor
口試委員
Advisory Committee
口試日期
Date of Exam
2016-07-22
繳交日期
Date of Submission
2016-08-29
關鍵字
Keywords
細胞黏附分子、血管內皮生長因子、子癲前症、白血球跨內皮細胞層移行、微陣列晶片
Cell adhesion molecules 1, Vascular endothelial growth factor 1, Leukocyte transendothelial migration, Preeclampsia, microarray analysis
統計
Statistics
本論文已被瀏覽 5680 次,被下載 55
The thesis/dissertation has been browsed 5680 times, has been downloaded 55 times.
中文摘要
子癲前症在台灣的發生率約2-5%,比國外5-7%低,但它卻是造成台灣地區孕婦死亡的一個主要原因。造成子癲前症的發病機轉至今仍無定論,但是一般認為與胎盤異常有關造成胎盤功能降低導致胎盤缺血和全身血管痙攣。據推測,子癲前症可以分為兩個階段:滋養層細胞受某些原因的影響使侵入母體蛻膜細胞的能力較差,因此血管維持高張力造成阻力提高,造成胎盤功能降低而產生血管內皮細胞功能喪失、血栓、血小板聚集與胎盤功能降低導致胎盤缺血及氧化壓力等一系列反應,則供應胎兒的血液與養分含量也會大幅減少;缺血和氧化壓力會促使胎盤分泌促進發炎的細胞激素、脂質過氧化反應的產物與滋養層細胞碎片進入母體內循環,進而誘發嚴重的母體全身性發炎反應以致於造成子癲前症的各種症狀。因此本研究使用微陣列晶片分析正常與子癲前症懷孕婦女胎盤中的基因差異表現,進而探討這些差異表現的基因是否與白血球跨內皮細胞層移行訊號傳遞路徑有關聯?實驗結果發現差異表現的基因VCAM-1、CADM-1在子癲前症組的表現都比正常懷孕組要高很多,這些細胞黏附分子可能是形成子癲前症過程的一種機制,可做為子癲前症發炎反應時所引發的白血球活化至內皮細胞損傷部位的標誌物。
Abstract
Preeclampsia (PE) incidence in Taiwan is 2-5%, less than the average rate of 5-7% worldwide; however, it is one of the three major causes of maternal death in Taiwan. Unfortunately, the cause of PE remains unclear. PE is associated with abnormal placentation, reduced placental perfusion and systemic vasospasm. It is assumed that PE can be divided into two stages: In the pre-clinical stage, insufficient trophoblast invasion in the first half of pregnancy results in poor placentation. Subsequent uteroplacental insufficiency leads to placental ischemia and oxidative stress. The ischemic and oxidatively stressed placenta releases pro- inflammatory cytokines, lipid peroxidation products and trophoblast debris into the maternal circulation, which in turn trigger an exaggerated maternal systemic inflammatory response as well as cause a variety of symptoms of PE. This study used microarray analysis of differential gene expression in human placentas from PE and normal pregnancies, and then investigate whether the differential gene expression connected with signal transduction of leukocyte transendothelial migration. The results showed that vascular endothelial growth factor 1 (VEGF-1) and cell adhesion molecules 1 (CADM-1) in PE group higher than the normal pregnancies group. The cell adhesion molecule may play a role in the pathophysiology of PE. VEGF-1 and CADM-1 expression are up-reaglated upon stimulation by inflammatory mediators, probably by contributing to the reduced trophoblast invasion and increased vascular damage. These may be useful biomarkers in predicting PE.
目次 Table of Contents
目 錄
論文審定書............................................................................................................... i
誌謝........................................................................................................................... ii
中文摘要................................................................................................................... iii
英文摘要................................................................................................................... iv
目錄........................................................................................................................... v
圖次........................................................................................................................... vii
表次........................................................................................................................... viii
第一章 緒論............................................................................................................. 1
1.1 子癲前症(Preeclampsia)的簡介................................................................ 1
1.2 子癲前症與白血球跨內皮細胞層移行訊號傳遞路徑的關聯...... 3
第二章 研究目的............................................................................. 5
第三章 實驗材料............................................................................. 6
3.1 人體胎盤檢體收集.............................................................. 6
3.2 胎盤組織均質..................................................................... 6
3.3 核醣核酸萃取與定量.......................................................... 6
3.4 基因晶片上機前處理.......................................................... 6
3.5 基因晶片掃描..................................................................... 7
3.6 基因晶片圖譜分析............................................................... 7
3.7 反轉錄-聚合酶反應(RT-PCR)................................................ 7
3.8 即時定量聚合酶連鎖反應(Quantitative Real-time RT-PCR)............ 7
第四章 實驗方法.............................................................................. 8
4.1 子癲前症懷孕孕婦與對照組正常懷孕孕婦臨床檢體胎盤收集流程.... 8
4.2 胎盤組織均................................................................................................ 8
4.3 核醣核酸的萃取與定量............................................................................ 9
4.4 基因晶片上機前處理............................................................ 9
4.5 基因表現圖譜掃描與分析........................................................ 10
4.6 即時定量聚合酶連鎖反應(Quantitative Real-time RT-PCR).................. 10
4.7 統計分析.................................................................................................... 12
第五章 實驗結果..................................................................................................... 13
5.1子癲前症懷孕孕婦與對照組正常懷孕孕婦臨床檢體資料.................... 13
5.2 基因晶片的分析資料................................................................................ 14
5.3 使用即時定量聚合酶連鎖反應來驗證基因晶片的結果........................ 15
第六章 討論............................................................................................................. 16
參考文獻................................................................................................................... 18

圖 次
圖一、 正常懷孕與子癲前症子宮螺旋動脈的變化............................................... 20
圖二、 子癲前症的兩個發展階段.......................................................................... 21
圖三、 白血球與血管內皮細胞間的互動示意圖.................................................. 22
圖四、 主成份分析圖(PCA).................................................................................... 23
圖五、 微陣列晶片差異表現基因的熱力圖(heatmap).......................................... 24
圖六、 白血球跨內皮細胞層移行訊號傳遞路徑.................................... 25
圖七、 CADM-1的基因表現.................................................................................. 26
圖八、 CLDN3的基因表現..................................................................................... 27
圖九、 CLDN8的基因表現..................................................................................... 28
圖十、 VCAM-1的基因表現................................................................................... 29

表 次
表一、 基因晶片分析孕婦與胎兒的臨床特徵....................................................... 30
表二、 孕婦與胎兒的臨床特徵............................................................................... 31
表三、106個差異表現的基因名單......................................................................... 32
表四、 正常與子癲前症差異表現基因參與的訊號傳遞路徑............................... 36
表五、 使用在即時定量聚合酶連鎖反應的引子名單.......................................... 37
參考文獻 References
1. Zhang HH, Wang YP, Chen DB. Analysis of Nitroso-Proteomes in Normotensive and Severe Preeclamptic Human Placentas.
Biol Reprod. 2011; 84(5): 966-75.

2. Redman CW, Sargent IL. Latest Advances in Understanding Preeclampsia. Science. 2005;308(5728):1592-4.

3. Cuckle H, von Dadelszen P, Ghidini A. Current controversies in prenatal diagnosis 4: pregnancy complications due to placental
vascular disease (pre- eclampsia, FGR): are we ready for prevention? Prenat Diagn. 2013; 33(1): 17-20.

4. Molvarec A, Shiozaki A, Ito M, Toldi G, Stenczer B, Szarka A, Nakashima A, Vásárhelyi B, Rigó J Jr, Saito S. Increased
prevalence of peripheral blood granulysin-producing cytotoxic T lymphocytes in preeclampsia. J Reprod Immunol. 2011; 91
(1-2): 56-63.

5. Saito S, Umekage H, Sakamoto Y, Sakai M, Tanebe K, Sasaki Y, Morikawa H. Increased T-helper-1-type immunity and
decreased T-helper-2-type immunity in patients with preeclampsia. Am J Reprod Immunol. 1999;41(5): 297-306.

6. Toder V, Blank M, Gleicher N, Voljovich I, Mashiah S, Nebel L. Activity of natural killer cells in normal pregnancy and edema-
proteinuria-hypertension gestosis. Am J Obstet Gynecol. 1983;145(1): 7-10.

7. Darmochwal-Kolarz D, Leszczynska-Gorzelak B, Rolinski J, Oleszczuk J. The expression and concentrations of Fas/APO-1
(CD95) antigen in patients with severe pre-eclampsia. J Reprod Immunol. 2001;49(2): 153-64.

8. Walsh SW. Plasma from preeclamptic women stimulates transendothelial migration of neutrophils. Reprod Sci. 2009;16(3):
320-5.

9. Fotis L, Giannakopoulos D, Stamogiannou L, Xatzipsalti M. Intercellular cell adhesion molecule-1 and vascular cell adhesion
molecule-1 in children. Do they play a role in the progression of atherosclerosis? Hormones. 2012;11(2):140-6.

10. Budak E, Madazli R, Aksu MF, Benian A, Gezer A, Palit N, Yildizfer F. Vascular cell adhesion molecule-1 (VCAM-1) and
leukocyte activation in pre-eclampsia and eclampsia. Int J Gynaecol Obstet. 1998;63(2):115-21.

11. Djurovic S, Schjetlein R, Wislaff F, Haugen G, Berg K. Increased levels of intercellular adhesion molecules and vascular cell
adhesion molecules in preeclampsia. Br J Obstet Gynaecol. 1997;104(4):466-70.

12. Coata G, Pennacchi L, Bini V, Liotta L, Di Renzo GC. Soluble adhesion molecules: marker of preeclampsia and intrauterine
growth restriction. J Matern Fetal Neonatal Med. 2002;12(1):28-34.
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