臺灣博碩士論文加值系統

日本腦炎病毒會感染人類的中樞神經，並導致急性的腦炎症狀，去髓鞘作用是其明顯病徵之一。日本腦炎病毒誘發去髓鞘作用的機制目前仍不清楚，但多數的研究指出病毒可能直接對中樞神經細胞造成損傷，而免疫反應間接對中樞神經細胞造成影響也可能與去髓鞘發生有關。因此，了解中樞神經中可被病毒感染的細胞種類相當重要，本研究希望了解日本腦炎病毒感染各神經細胞的情形，同時探討被感染的細胞與去髓鞘作用的關係。我們利用尾靜脈注射，觀察感染日本腦炎病毒的鼠腦組織，藉電子顯微鏡和免疫組織染色觀察所提供的證據，以了解這些被病毒侵犯的神經細胞是否與神經性損傷有關。結果顯示，具有產生髓鞘功能的寡突觸細胞似無法被病毒感染，反之，其它中樞神經細胞則常見被病毒感染的情形。被病毒感染的神經元有趨向細胞壞死的現象，另一方面，腦組織中也可發現有些細胞正在進行細胞凋亡，若利用TUNEL進行檢測，發現這些細胞包括許多星狀細胞和極少數的寡突觸細胞。因此中樞神經的去髓鞘現象，有可能直接和病毒感染神經元或星狀細胞有關，但與寡突觸細胞並無密切的關係。因細胞凋亡和腦炎的嚴重性有關，據推測其在日本腦炎病毒引發的去髓鞘作用中，可能也扮演重要的角色。

Japanese encephalitis virus (JEV) causes severe central nervous system (CNS) infection in humans and produces acute encephalitis, resulting in inflammatory demyelination. The mechanisms leading to JEV-induced demyelination are still under study but most likely involve either virus induced or immune-mediated damage or both to cells in the CNS. It is therefore important to identify the type of cells in which continued virus production is permitted. In this study, we evaluated the possibility of virus infection in neuronal cells and its relation with occurrence of demyelination. As evidenced by electron microscopy and immunohistochemistry, we extended the study to CNS tissues from mice suffering from JEV-induced disease by intravascular inoculation. We then assessed the role of virus presented in these cells in causing true neuronal injury. Our findings showed that the main cell for viral replication in the CNS was not oligodendrocytes that produce and extend myelin to wrap the axon. On the other hand, virus-infected neurons were frequently seen in the CNS. Moreover, the necrotic death was usually shown in infected neurons. Apoptosis was also observed in certain cells in JEV-positive brains. TUNEL-positive cells were identified as mainly astrocytes, and few oligodendrocytes on the basis of immunolabling with cell-specific markers. Most of demyelination presented in the CNS, therefore, is probably the result of neuron damage due to viral infection of neuronal cells, and astrocytes as well, but probably not oligodendrocytes. Because apoptosis of neural cells may give rise to aggravated encephalopathy, it may also play an important role in JEV-induced demyelination.

第一章 文獻回顧
第一節 日本腦炎病毒及其流行病學
一、 日本腦炎病毒的傳播………………………………………….…1
二、 日本腦炎的臨床症狀………………………………………….…2
三、 日本腦炎病毒顆粒構造……………………………………….…2
第二節 日本腦炎病毒的感染模式及傳遞
一、 日本腦炎病毒的感染模式……………………………………….4
二、 日本腦炎病毒於中樞神經的傳遞……………………………….5
第三節 中樞神經的特性與細微結構
一、 中樞神經細胞的特性…………………………………………….7
二、 中樞神經細胞的功能……………………………………………10
第四節 病毒與中樞神經的關係
一、 病毒與腦炎的關係………………………………………………13
二、 病毒對神經細胞的影響…………………………………………13
第二章 研究目的………………………………………………………………..16
第三章 材料與方法
第一節 實驗材料…………………………………………………………..18
第二節 實驗方法…………………………………………………………..20
第四章 結果
一、 染日本腦炎病毒對腦組織的影響…………………………………..27
二、 感染日本腦炎病毒的細胞種類……………………………………..28
三、 日本腦炎病毒對神經細胞的影響…………………………………..30
第五章 討論……………………………………………………………………..32
參考文獻…………………………………………………………………………38
附錄………………………………………………………………………………47

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