臺灣博碩士論文加值系統

摘要
截至目前為止的研究，我們已經證明在大白鼠乳腺肌上皮細胞上有類似CD200的表現，且不同生理狀況下的乳腺可產生不同的賀爾蒙去影響管腔上皮細胞變化，肌上皮細胞則隨著上皮細胞之變化而改變。的確，我們目前的研究已進一步發現在大白鼠乳腺休止、懷孕、泌乳及離乳四個生理時期，其乳腺肌上皮細胞的CD200均會有不同表現。於大白鼠乳腺休止期，CD200會分佈在肌上皮細胞的整個細胞膜上。但在懷孕期，CD200的分佈卻有所改變，此時CD200僅表現在肌上皮細胞靠近基底膜側的細胞膜上；靠近上皮細胞側細胞膜上的CD200卻消失。進入泌乳期，CD200在肌上皮細胞細胞膜上兩側的表現剛好和懷孕期的表現相反。有趣的是，離乳期的乳腺上，肌上皮細胞會包被正在退化萎縮的腺泡，且其靠近基底膜側細胞膜上的CD200免疫反應是中等程度且連續的分佈；然而靠近已經退化上皮細胞那側的細胞膜，CD200呈現較為強烈且偶發性的免疫反應。進一歩研究乳腺肌上皮細胞CD200在離乳期不同時間點的變化，證據顯示肌上皮細胞CD200的表現明顯增加。隨著乳腺退化的進行，肌上皮細胞會將其含有愈多明顯凸出物的突起縮回。在大白鼠乳腺離乳晚期，其管道及腺泡均退化至類似乳腺休止時期之結構，此時，強烈的CD200免疫反應不僅出現在肌上皮細胞上，同時亦出現在部分的上皮細胞上。而青春期五週大之大白鼠乳腺上皮細胞也被證實有明顯CD200的免疫反應，強烈的CD200免疫反應表現在五週大大白鼠乳腺大部分但非全部的上皮細胞之細胞膜及其頂端修飾物結構上。有趣的是，構成末端盲管芽最外層的帽細胞，卻一直沒有CD200的免疫反應。同樣的現象亦發生在大白鼠乳腺懷孕、泌乳及離乳期的上皮細胞中。本實驗結果指出，由於賀爾蒙環境的刺激造成乳腺上皮細胞有不同的生理狀態，乳腺肌上皮細胞可能會隨著上皮細胞不同的生理狀況而有所改變，同時，乳腺的生長可能會透過CD200黏著分子。

Abstract
In the present study, we demonstrated a similar CD200 expression in the myoepithelial cells of the mammary glands. Myoepithelial cells were well known to be altered morphologically under different hormonal conditions in a physiologic state correlative to the luminal epithelial cells. Indeed, our present study further showed a differential expression of myoepithelial CD200 in the rat mammary gland during resting, pregnancy, lactation and involution. CD200 is distributed at entire cytoplasmic membrane of myoepithelial cells in the virgin mammary gland. This expression is altered when rats are in pregnancy and myoepithelial CD200 is expressed only on the cytoplasmic membrane facing the basal lamina and absent on that facing the epithelium. The polarity of CD200 expression on myoepithelial cells is reversed during lactation. Interestingly, myoepithelial cells investing regressing alveoli of involuted glands showed a moderate but continuous CD200 immunoreactivity on the cytoplasmic membrane facing the basal lamina. Opposite degenerated epithelial cells, the immunoreactivity on the membrane is however intense but sporadic. Extended study on chronological changes of myoepithelial CD200 in the involuted glands showed an increased CD200 expression on myoepithelia that retracted their processes bearing more obvious protrusions with advance of the involution. At the final phase of mammary involution when the ducts and alveoli regressed towards a resting state, intense CD200 immunoreactivity was found at not only myoepithelia but some epithelial cells. The remarkable expression of epithelial CD200 was also evidenced in the mammary glands of 5-week-old pubertal rats that showed intense immunoreactivity on the cytoplasmic membrane including its apical modifications of major but not all epithelial cells. Interestingly, the outermost layer made up by cap cells of the terminal end buds has nerve possessed CD200. It is also true for epithelial cells of mammary glands during pregnancy, lactation and involution. These results indicated that myoepithelial cells of the mammary glands may be changed and related to the physiologic state of the epithelial cells by hormonal environments synchronizing the gland growth through in part CD200 adhesion molecules.

目錄 頁次
目錄.................................................................I
圖目錄..............................................................IV
附圖目錄............................................................VI
中文摘要...........................................................VII
英文摘要............................................................IX
第一章 緒論...........................................................1
第一節 CD200.......................................................1
壹、CD200的起源...................................................1
貳、CD200之相關研究...............................................2
第二節 大白鼠乳腺構造................................................4
壹、大體解剖構造 (Gross anatomy)..................................4
貳、顯微解剖構造 (Microscopic anatomy)............................5
第二章 研究目的......................................................20
第三章 材料與方法....................................................21
第一節 實驗材料....................................................21
壹、實驗動物.....................................................21
貳、實驗藥品.....................................................21
參、免疫組織化學染色抗體..........................................21
肆、免疫螢光染色抗體..............................................21
伍、其他化學試劑.................................................22
陸、儀器設備....................................................23
第二節 實驗設計與研究方法...........................................23
壹、實驗設計、動物犧牲與標本組織取得................................23
貳、冷凍切片之標本備製與切片.......................................26
參、免疫組織化學染色..............................................27
肆、免疫組織化學染色之對比染色及脫水................................28
伍、免疫螢光染色.................................................28
陸、免疫螢光染色之對比染色及脫水...................................29
柒、電子顯微鏡切片之標本備製與切片..................................29
第四章 結果..........................................................31
第一節 運用CD200免疫組織化學染色法，探討CD200在大白鼠乳腺休止、懷孕、泌乳
及離乳四個生理時期之分佈......................................31
壹、光學顯微鏡觀察大白鼠乳腺在休止、懷孕、泌乳及離乳時期，CD200之分佈..31
貳、電子顯微鏡觀察大白鼠乳腺在休止、懷孕、泌乳及離乳時期，CD200之分佈..33
第二節 運用CD200免疫組織化學染色法，進一步探討大白鼠乳腺離乳期，CD200在乳
腺肌上皮細胞之分佈............................................35
壹、離乳二天的大白鼠乳腺上CD200之分佈..............................35
貳、離乳四天的大白鼠乳腺上CD200之分佈..............................36
參、離乳十四天的大白鼠乳腺上CD200之分佈.............................37
肆、離乳二十一天的大白鼠乳腺上CD200之分佈...........................37
第三節 運用CD200免疫組織化學染色法，進一步探討五週大大白鼠乳腺休止期，CD200
於乳腺末端盲管芽、小泡芽及終末導管之分佈........................38
壹、光學顯微鏡觀察五週大的大白鼠乳腺休止期CD200之分佈................39
貳、共軛焦顯微鏡觀察五週大的大白鼠乳腺休止期CD200之分佈...............40
第五章 討論..........................................................42
第一節 乳腺肌上皮細胞CD200之顯著變化，除可反應出肌上皮細胞形態上的改變外，
亦代表其不同的功能(生理)階段...................................42
第二節 乳腺肌上皮細胞與腺體上皮細胞的前軀細胞均可被CD200標示上，且二者可透
過CD200間同質性黏著的結合方式，來達成細胞與細胞間的交互作用.......46
第三節 CD200是已分化的乳腺腺體組織當中肌上皮細胞的專一標誌..............47
第四節 大白鼠乳腺離乳期CD200所扮演的角色..............................47
壹、大白鼠乳腺離乳期是一種免疫反應的表現............................47
貳、CD200為一種免疫抑制分子，其接受體存在於巨噬細胞表面...............50
參、大白鼠乳腺離乳期，肌上皮細胞不被巨噬細胞吞噬，亦不會引發更嚴重的自體
免疫反應.....................................................52
第六章 結論..........................................................54
參考文獻...........................................................93

圖目錄 頁次
圖一、光學顯微鏡觀察成熟(八週大)大白鼠乳腺休止期CD200免疫組織化學染色之表現.56
圖二、光學顯微鏡觀察大白鼠乳腺懷孕期CD200免疫組織化學染色之表現............58
圖三、光學顯微鏡觀察大白鼠乳腺泌乳期CD200免疫組織化學染色之表現............60
圖四、光學顯微鏡觀察離乳七天的大白鼠乳腺上CD200免疫組織化學染色之表現......62
圖五、電子顯微鏡觀察成熟(八週大)大白鼠乳腺休止期CD200免疫組織化學染色之表現.64
圖六、電子顯微鏡觀察大白鼠乳腺懷孕期CD200免疫組織化學染色之表現............66
圖七、電子顯微鏡觀察大白鼠乳腺泌乳期CD200免疫組織化學染色之表現............68
圖八、電子顯微鏡觀察離乳七天的大白鼠乳腺上CD200免疫組織化學染色之表現......70
圖九、光學顯微鏡觀察離乳二天的大白鼠乳腺上CD200免疫組織化學染色之表現......72
圖十、光學顯微鏡觀察離乳四天的大白鼠乳腺上CD200免疫組織化學染色之表現......74
圖十一、光學顯微鏡觀察離乳十四天的大白鼠乳腺上CD200免疫組織化學染色之表現...76
圖十二、光學顯微鏡觀察離乳二十一天的大白鼠乳腺上CD200免疫組織化學染色之表現.78
圖十三、光學顯微鏡觀察五週大之大白鼠乳腺休止期CD200免疫組織化學染色之表現...80
圖十四、光學顯微鏡觀察五週大之大白鼠乳腺休止期，CD200在末端盲管芽、小泡芽及
終末導管免疫組織化學染色之表現..................................82
圖十五、共軛焦顯微鏡觀察五週大之大白鼠乳腺休止期，CD200在末端盲管芽、小泡芽
及終末導管免疫組織化學染色之表現................................84
圖十六、大白鼠乳腺在休止、懷孕、泌乳及離乳時期，CD200分佈在肌上皮細胞之模式
圖...........................................................86

附圖目錄 頁次
附圖一、雌性小鼠乳腺分佈圖.............................................88
附圖二、末端盲管芽和小泡芽之構造模式圖..................................88
附圖三、56天大雌性大白鼠其乳腺休止期第四對乳腺之模式圖....................90
附圖四、活躍期之乳腺構造..............................................90
附圖五、大白鼠乳腺不同生理階段之肌上皮細胞利用免疫組織化學染色法觀察之模式圖.92

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