臺灣博碩士論文加值系統

　　腸病毒71型感染後的臨床症狀通常是輕微的，但有時在嬰幼兒感染會產生併發症如無菌性腦膜炎、類小兒麻痺脊髓炎和腦膜炎，更嚴重的會出現肺水腫及心臟衰竭等症狀。我們的研究是希望藉由小鼠感染模式，探討中樞神經在腸病毒71型感染時所引起的疾病與肺部異常的關係，並嘗試了解可能的細胞或分子機制。我們以顱內注射的方式感染腸病毒71型小鼠適應株－MP4至14日齡的ICR小鼠，在感染後第五天，小鼠開始出現後肢麻痺的現象，甚至會死亡。病毒的定量分析發現在腦幹及小腦區域的病毒量比大腦要高，因此我們推測這兩個區域可能為腸病毒71型在感染中樞神經時的主要目標。腦組織切片的免疫組織化學染色發現，在橋腦的區域會產生catecholamine的神經細胞有減少的趨勢；肺部的病理切片顯示小鼠在感染腸病毒後會有肺氣腫的現象，且後肢肌肉及肋間肌的組織切片都有嚴重的組織壞死現象。以Whole Body Plethysmograph分析，發現受到腸病毒71型感染的小鼠比起對照組會有明顯肺功能指數下降的現象，包括最大吸氣值、最大呼氣值、潮氣容積以及每分鐘的呼吸容積，但以顱內注射酯多醣體的小鼠則不會出現肺功能異常的變化。其他的感染途徑如肌肉注射或口內餵食感染，同樣會造成小鼠感染並出現肺功能的異常，但是發病的時間與中樞神經被波及有關。整體而言，腸病毒71型感染小鼠所引起的肺功能異常與中樞神經感染有著一定的關係，且catecholamine也可能是參與其中的調控。

　　Enterovirus 71 (EV71) infection is usually mild, but occasionally leads to severe diseases such as aseptic meningitis, polio-like myelitis, encephalitis, pulmonary edema and cardiac insufficiency in neonates. This study is attempted to clarify the relationship between EV71-induced central nervous system (CNS) infection and pulmonary abnormalities, and the underlying cellular and molecular mechanism using an experimental infection model. Fourteen-day-old ICR mice developed hindlimb paralysis 5 days followed by death after intracranial inoculation of EV71/MP4. The brain stem and cerebellum had a higher viral load than the cerebrum suggesting it might be the major target site for EV71 in the CNS. The lung displayed emphysema and there were severe necrosis in the hindlimb and intercostals muscle. The whole body plethysmograph analysis indicated that the EV71-inoculated but not LPS-inoculated mice had a marked alteration in pulmonary function parameters including peak inspiratory flow, peak expiratory flow, minute volume and tidal volume when compared to mock-infected mice. Other infectious routes including intramuscular and oral inoculation also could induce lung function abnormality but with a delay onset. Immunohistochemical staining showed the numbers of catecholaminergic neurons in the midbrain were gradually declined after infection. These results suggest that there is a relationship between the CNS disease and pulmonary abnormality in EV71-infected mice and that catecholamine expression level might be involved in the disease process.

目錄

中文摘要 Ⅰ
英文摘要 Ⅱ
目錄 Ⅲ
圖目錄 Ⅴ
第一章 緒論 1
第二章 研究目標與實驗設計 10
第三章 材料與方法 12
A 材料 12
A-1 實驗動物 12
A-2 細胞株 12
A-3 病毒 12
A-4 藥品與試劑 12
A-5 抗體與試劑組 14
A-6 耗材 14
A-7 儀器 15
B 方法 16
(A) 細胞培養 16
(B) 病毒培養與感染 19
(C) 組織切片與染色觀察 23
(D) ELISA:細胞激素(cytokines)的監測 28
(E) 肺功能的測定:Unrestrained Whole Body Plethysmography 30
第四章 結果 32
第五章 討論 37
第六章 參考文獻 43
圖 50

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