出版年:

2001

研究生:

郭雅純

研究生(英文姓名):

論文名稱:

探討鼠腦部位在致死性HSV腦炎的重要性

英文論文名稱:

Mapping the sites in mouse brain critical for lethal HSV encephalitis

指導教授:

陳舜華博士

學位類別:

碩士

校院名稱:

國立成功大學　

系所名稱:

微生物暨免疫學研究所

學號:

S46881123

學年度:

89

語文別:

中文

論文頁數:

0

關鍵詞:

單純疹病毒第一型 ; 腦炎

英文關鍵詞:

HSV ; encephalitis

被引用次數:

0

[ 摘要 ]

單純疹病毒第一型（HSV-1）可以在哺乳類動物的神經系統中複製與擴散，並造成致死性腦炎。以前在人類病例或動物實驗的研究中，對於單純疹病毒在腦炎時在腦的分布與生長複製情形及造成宿主死亡的原因，並無清楚的闡釋。在這個研究裡，我觀察病毒在鼠腦內複製生長的區域及程度，並試圖決定病毒腦炎造成宿主死亡的區域。首先，小鼠經眼睛感染後，在不同的時間點將各部分鼠腦分離出，測其病毒量。在嗅腦、大腦皮層、海馬迴、小腦及腦幹中皆可發現具感染力的病毒，其中以大腦皮層與腦幹中所含病毒量特別地高。此外，我們也使用顱內立體定位注射方法將病毒注射至鼠腦內病毒複製的區域，以檢視這些區域對於病毒腦炎之重要性。我挑選了腦幹與海馬迴來作比較。令人訝異地，海馬迴與腦幹的百分之五十致死率的病毒劑量並沒有顯著的差異性。而將顱內立體定位注射後的鼠腦分離並進行病毒計量後，我們看到病毒可以在注射的位置大量地複製, 但同時它們亦可以快速地散播至其它非注射區域，這樣的情況在海馬迴感染的組別裡尤其明顯。顱內立體定位注射後鼠腦的損傷則在PI染色或Annexin V與PI雙染色後，以流體細胞儀分析。無論是從腦幹或海馬迴感染，鼠腦的腦幹皆有超過百分之十的損傷死亡；但在海馬迴感染的組別裡，海馬迴所受損傷則不明顯。這些腦細胞死亡是因為細胞凋亡。所以，海馬迴及腦幹的百分之五十致死率差異不明顯可能是因在腦幹裡的腦細胞受到損傷所致。

[ 英文摘要 ]

Herpes simplex virus 1 (HSV-1) multiplies and spreads in mammalian nervous system to cause lethal encephalitis. Previous postmortem histopathological examination of human encephalitis cases and mouse studies did not clearly demonstrate how HSV replicates and kills host during encephalitis. In this study, we characterized the sites viruses multiply in mouse brain and whether any of these sites is critical for causing death after infection. We applied titration method to determine the distribution and growth of virus in various parts of mouse brain at different time points after infection. Infectious virus was recovered from olfactory bulbs, frontal cortex, hippocampus, cerebellum and brain stem. Virus titers were very high in frontal cortex and brain stem. Meanwhile, stereotaxic injection of virus into replication sites was performed to investigate whether any of these sites is critical for lethal encephalitis. The brain stem and hippocampus were chosen for microinjection to compare the sensitivity to HSV-1in different regions of mouse brain. Surprisingly, LD50s of hippocampus and the nucleus tractus solitarius of brain stem are comparable. Titration of dissected brain tissues after stereotaxic microinjection showed that viruses replicate abundantly in injection sites, but they also can spread out to other non-injecting sites, particularly by hippocampus injection. The damage of mouse brain after microinjection was examined by flow cytometry using PI stain or Annexin V plus PI stain. More than 10 percent of cells in brain stem were damaged by both brain stem and hippocampus injection, while the damage in hippocampus was not significant even by hippocampus injection. Neuron damage was caused by apoptosis. Thus, comparable LD50s in hippocampus and brain stem may result from the damage of cells in brain stem.