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研究生:黃文翰
研究生(外文):Boon-Han Ng
論文名稱:大白鼠最後區經脂多醣類處理後其神經元及神經膠細胞的反應
論文名稱(外文):Response of neurons and glia in the area postrema of adult rats following lipopolysaccharide challenge
指導教授:溫振源溫振源引用關係
學位類別:碩士
校院名稱:國立臺灣大學
系所名稱:解剖學研究所
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:2004
畢業學年度:92
語文別:中文
論文頁數:49
中文關鍵詞:免疫分子最後區脂多醣類
外文關鍵詞:lipopolysaccharideimmunomoleculearea postrema
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最後區為環腦室周圍器官之一,其在內的微血管由穿孔性內皮細胞構成,因此不具有血腦障壁的保護弁遄C一般認為最後區可探測血液中有害物質,進而引發嘔吐反應來排除毒物。本實驗之目的是以免疫組織化學染色觀察正常生理狀態下,最後區內某特殊神經元及神經膠細胞免疫分子的分佈。第二部份試驗則是給予大白鼠靜脈注射脂多醣類,然後觀察最後區內神經元及神經膠細胞的早期急性反應。

實驗結果顯示在正常生理狀態下,最後區內具神經性一氧化氮的神經元之分佈不具有區域的差異性。而最後區內星狀神經膠細胞所表現的神經膠質纖維酸性蛋白則大部份出現在最後區背面及中間部位。最後區內巨噬細胞/小神經膠細胞會因其表現不同的免疫分子而有不同的分佈區域,例如具補體第三型受器巨噬細胞/小神經膠細胞主要分佈在最後區的腹面而僅稀疏地散佈在背面,具主要組織相容複合物第二型抗原的巨噬細胞/小神經膠細胞大部份則出現在最後區背面,而具未知細胞質或溶菌體抗原的巨噬細胞/小神經膠細胞主要分布在最後區的中間部位。

給予脂多醣類刺激後,最後區內c-Fos免疫活性迅速升高,而具神經性一氧化氮合成酶神經元在某些特殊時間點上有下降的趨勢。雙重免疫染色結果顯示c-Fos與具神經性一氧化氮合成酶神經元的關連性並不大。另外經脂多醣類刺激後,最後區內同時表現c-Fos與酪胺酸水解酶免疫活性的神經元明顯增加。靜脈注射脂多醣類後發現,最後區具不同免疫分子的神經膠細胞有不同反應趨勢。其中具神經膠質纖維酸性蛋白星狀神經膠細胞與具補體第三型受器巨噬細胞/小神經膠細胞都有早期急速下降的情形,然後才上升。而具主要組織相容複合物第二型抗原巨噬細胞/小神經膠細胞則隨時間增長而增高免疫活性,具未知細胞質或溶菌體抗原的巨噬細胞/小神經膠細胞則無明顯的變化。

綜合以上結果發現,在正常生理情況下,最後區內星狀神經膠細胞及巨噬細胞/小神經膠細胞所表現的不同免疫分子各有其分佈上的特異性,以因應最後區沒有血腦障壁保護的特性。當受到外來致病原刺激後,最後區內星狀神經膠細胞及巨噬細胞/小神經膠細胞所表現的不同免疫分子,各有其不同的表現形式,顯示作為中樞神經系統對外的窗口—最後區受到病原體刺激時會產生敏感且獨特的急性反應。
The area postrema (AP) is located on the dorsal surface of the medulla oblongata at the caudal end of the fourth ventricle. It is one of the so-called circumventricular organs (CVOs) that serve as an interface between the brain parenchyma and the cerebrospinal fluid (CSF)-containing ventricles. Because for its capaillary constrution by fenestrated endothelial cell, the most striking feature of the AP, in common with other CVOs, is its lack of a specific blood-brain barrier to large polar molecules. The major functional of the AP is to detect toxic material in the blood and to exclude the toxic substance by vomiting.

The aim of this study was to use immunohistochemical method to find out the normal distribution of some specific neurons, neuroglia and macrophages/microglia expressing different immunomolecules in the AP. The further study is to analyze acute responses of the above-mentioned cell elements in the AP following an intravenous injection (i.v.) of lipopolysaccharide (LPS).

The present results showed that neurons bearing neuronal nitric oxide synthase (nNOS) have no specific distribution in AP. However, glial fibrillary acidic protein (GFAP) expressed by astrocytes was heavily distributed at the dorsal and central portions of the AP. The AP macrophages/microglia expressing complement type 3 receptors were localized mainly in the ventral portion of the AP. Another AP macrophages/microglia that expressed the major histocompatibility complex class II antigen were frequently observed in the dorsal portion of the AP. Those with unknown cytoplasmic/lysosomal antigen (ED1) were localized mainly in the medial portion of the AP.

When challenged with LPS, the c-Fos containing neurons in the AP increased in number dramatically. The decreased expression of nNOS was found at specific time intervals after LPS treatment and not correlated to the Fos expression. However, AP neurons co-expressing with c-Fos and tyrosine hydroxylase were increased after treatment with LPS.

LPS treatment elicited different patterns of acute responses in AP macrophages/microglia bearing different immunomolecule. Macrophages/microglia possessing complement type 3 receptors and astrocytes showed an initial rapid drop of the immunoreactivity after LPS challenge but increased their reactivities thereafter. LPS treatment also resulted in a time-dependent increase of MHC class II antigen in AP macrophages/microglia. While, ED1 containing macrophages/microglia were not affected when challenged with LPS.

In conclusion, the expression of immunoreactive molecules in AP glial cell varies dependent on topographical distributions of the cells. This may be linked to the
lack of BBB feature in the AP. LPS-elicited different patterns in acute responses of AP astrocytes and macrophages/microglia containing different immunomolecule implicated that AP, as a window of CNS, would show a sensitive and special acute response to the pathogens.
目錄

中文摘要…………………………………………………………………I

Abstract………………………………………………………………III

第一章 引言

一、 文獻回顧

第一節 最後區………………………………………………………1

第二節 脂多醣類……………………………………………………5

第三節 脂多醣類對中樞神經系統內小神經膠細胞及星狀神經
膠細胞的影響………………………………………………7

第四節 脂多醣類對中樞神經系統神經元的影響…………………9

二、 研究目的……………………………………………………11

第二章 實驗材料與方法……………………………………………12

第三章 實驗結果

第一部份:正常大白鼠腦幹最後區內表現不同免疫分子之巨噬
細胞/小神經膠細胞及星狀神經膠細胞的分佈…………16

第二部份:大白鼠施以脂多醣類靜脈注射後,最後區內巨噬細胞/
小神經膠細胞的免疫分子及星狀神經膠細胞之變……17

第三部份:注射脂多醣類後,大白鼠最後區內神經性一氧化氮
合成酶與Fos表現的變化…………………………………19

第四部份:注射脂多醣類後,引發最後區內表現c-Fos的神經元屬性………………………………………………………………………20

第四章 討論

第一節 在正常情況下,最後區中具神經性一氧化氮
合成酶神經元及酪胺酸水解酶神經元之分佈………………………22

第二節 脂多醣類刺激下,最後區內神經性一氧化氮合成酶
表現之變化及此變化與c-Fos表現的相關性…………………………23

第三節 在脂多醣類刺激下,最後區內酪胺酸水解酶之表現
與c-Fos表現的相關性…………………………………………………24

第四節 在正常情況下,最後區中星狀神經膠細胞之神經膠質
纖維酸性蛋白分佈……………………………………………………25

第五節 在脂多醣類刺激下,最後區內星狀神經膠細胞與
其突起之表現…………………………………………………………26

第六節 在正常情況下,最後區內巨噬細胞/小神經膠細胞
的免疫分子:補體第三型受器、主要組織相容複合物
第二型抗原及未知的細胞質或溶菌體抗原的分佈…………………27

第七節 在脂多醣類刺激下,最後區內巨噬細胞/小神經膠細胞
的免疫分子:補體第三型受器、主要組織相容複合物
第二型抗原及未知的細胞質或溶菌體抗原的表現…………………29

圖表說明………………………………………………………………33

參考文獻………………………………………………………………42
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