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研究生:劉秀莉
研究生(外文):Liu, shiow-li.
論文名稱:去除雌性素對小鼠子宮及陰道上皮細胞死亡之影響
論文名稱(外文):Effect of estrolgen wtithdrawal on epithelial cell death in mouse uterus and vagina
指導教授:陳麗玉陳麗玉引用關係
指導教授(外文):Lih-Yuh Chen
學位類別:碩士
校院名稱:國立成功大學
系所名稱:生理學研究所
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:1999
畢業學年度:87
語文別:中文
論文頁數:60
中文關鍵詞:雌性素子宮陰道上皮細胞死亡
外文關鍵詞:estrogenuterusvaginaepithelium cell death
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中文摘要
雌性素刺激子宮及陰道上皮細胞的增生,而切除卵巢後子宮及
陰道上皮細胞則會經由凋亡的方式而進行退化。目前已證實Bcl-2及Bcl-xL的表現,細胞能夠獲得保護而免於死亡。相反的,Bax及Bcl-xS則導致細胞死亡。本論文探討在雌性素的處理及去除雌性素的情況之下,對小鼠子宮及陰道上皮細胞的組織型態變化及控制細胞死亡的
Bcl-2 family表現之影響。將小鼠去除卵巢至少2週後,皮下植入E2 ( 17-Estradiol )膠囊,三天後將動物分成兩組:一組持續保留E2膠囊,另一組則去除E2膠囊,分別於不同時間犧牲動物,並取出子宮及陰道,利用TUNEL染色法及免疫組織化學染色法來分析。由實驗結果顯示,在雌性素的處理下,子宮管腔上皮細胞的厚度及面積明顯增加,陰道上皮細胞層的厚度及層數亦顯著增加。然而,去除雌性素之後,子宮管腔上皮細胞的厚度及面積下降,而陰道上皮細胞層的厚度及層數亦同樣會下降。在移除E2膠囊後的第一天至第五天,在子宮管腔上皮細胞及陰道上皮細胞的表皮層與基底層出現許多凋亡的細胞。另外在Bcl-2 family的表現方面,給予E2的處理後,Bcl-2於子宮管腔上皮細胞的表現微微增加,而在移除E2膠囊後的第三天至第七天,Bcl-2的表現會逐漸下降回到切除卵巢後未給予E2刺激時的程度。在雌性素的處理及去除雌性素的情況之下,Bax及Bcl-x的表現於子宮管腔上皮細胞並無任何變化。此外,於陰道的上皮細胞層亦偵測到Bcl-2 , Bax及Bcl-x的表現,但在雌性素的處理及去除雌性素的情況之下,這些蛋白質的表現並無任何明顯的變化。以上結果顯示,雖然去除荷爾蒙可導致子宮及陰道上皮細胞進行凋亡,但所測試Bcl-2 family之表現變化與凋亡細胞的產生並無甚大關聯。

Abstract
Estrogen stimulates the proliferation of epithelial cells in the uterus and the vagina. Ovariectomy induces regression via apoptotic death in these two organs. Bcl-2 and Bcl-xL are known to protect cell from apoptotic death, whereas Bax and Bcl-xS induce apoptotic death. This study has investigated the effect of estrogen treatment and its removal on histological change and the expression of the cell death regulatory proteins, Bcl-2 family, in mouse uterine and vaginal epithelium. Mice were ovariectomized for at least two weeks and then received subcutaneous implantation of capsules containing 17-estradiol (E2). After three days, animals were divided into two groups: one group were treated with estrogen continuously, and the E2 capsules were removed from the second group. Mice were sacrificed at various times and their uteri and vaginae were removed for immunohistochemical staining of Bcl-2, Bax and Bcl-x and for the detection of apoptotic cells by TUNEL. Our results showed that estrogen treatment increased the height and area of luminal epithelium in the uterus. It also increased the thickness and cell layers of vaginal epithelium. However, after the removal of E2 capsules, the height and area of uterine luminal epithelium as well as the thickness and the number of epithelial cell layers in the vagina were decreased. In addition, apoptotic cells appeared in the luminal epithelium of uterus, and in the superficial and basal layer of stratified epithelium of the vagina. The treatment of estrogen slightly increased the expression of Bcl-2 in luminal epithelium of the uterus, but the removal of hormone caused the reduction of Bcl-2 expression gradually back to the level of ovariectomized mice. Bax and Bcl-x were expressed in uterine luminal epithelium, but no changes in Bax and Bcl-x expression could be observed. Bcl-2, Bax and Bcl-x were also detected in vaginal epithelium. No obvious changes in the expression of these Bcl-2 family proteins could be observed. Our results suggest that hormone withdrawal induces apoptotic death in both uterine and vaginal epithelium, and that the changes in the expression of Bcl-2 family proteins are not correlated with epithelial cell death.

著作權聲明
誌謝 i
中文摘要 ii
英文摘要 iii
目錄 v
圖表目錄 vii
I. 緒論 1
II. 材料與方法 6
( I )方法
A. 實驗動物
B. 動物的處理
C. 17-Estradiol ( E2 )膠囊的製作
D. 組織形態學( Morphology )研究的製備
E. 石蠟切片之蘇木精及伊紅( Hematoxylin & Eosin stain )染色
F. 免疫組織化學染色法( Immunohistochemistry )
G. TUNEL stain
H. 子宮管腔上皮細胞厚度及陰道上皮細胞層厚度的測量方法
I. 子宮管腔上皮細胞面積的測量方法
J. 統計分析
( II )器材與試劑 19
III. 結果 22
一.子宮管腔上皮細胞在生長及退化過程中組織型態上之變化
二.陰道上皮細胞層在生長及退化過程中組織型態上之變化
三.去除E2對子宮管腔上皮細胞凋亡之影響
四.去除E2對陰道上皮細胞層細胞凋亡之影響
五.子宮與陰道上皮細胞在生長及退化過程中Bcl-2之變化
六.子宮與陰道上皮細胞在生長及退化過程中Bax之變化
七.子宮與陰道上皮細胞在生長及退化過程中Bcl-x之變化
IV. 討論 29
V. 參考資料 33
圖表目錄
圖1. 小鼠子宮管腔上皮細胞組織型態之變化
圖2. 小鼠子宮管腔上皮細胞的厚度及面積的變化
圖3. 小鼠陰道上皮細胞層及腳底皮膚組織型態之變化
圖4. 小鼠陰道上皮細胞層的厚度及層次的變化
圖5. 利用TUNEL染色法偵測去除E2後子宮管腔上皮細胞凋亡的情形
圖6. 利用TUNEL染色法偵測去除E2後陰道上皮細胞層內皮細胞凋亡的情形
圖7. 利用免疫組織化學染色法觀察子宮管腔上皮細胞中Bcl-2表現的變化
圖8. 利用免疫組織化學染色法觀察陰道上皮細胞層內Bcl-2表現的變化
圖9. 利用免疫組織化學染色法觀察子宮管腔上皮細胞中Bax表現的變化
圖10. 利用免疫組織化學染色法觀察陰道上皮細胞層內Bax表現的變化
圖11. 利用免疫組織化學染色法觀察子宮管腔上皮細胞中Bcl-x表現的變化
圖12. 利用免疫組織化學染色法觀察陰道上皮細胞層內Bcl-x表現的變化

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