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標題: 探討第一型干擾素訊號路徑在日本腦炎病毒感染大鼠神經膠細胞所扮演之角色
Role of Type I Interferons Signaling Pathway during Japanese encephalitis virus Infection in Rat Glial Cell
作者: 周明論
Chou, Ming-Lun
關鍵字: Japanese encephalitis virus;日本腦炎病毒;Rat cerebral cortex primary cell;Type I interferon;Signal transducer and activator of transcription;Interferon stimulated gene;大鼠大腦皮質初代細胞;第一型干擾素;訊號轉導與轉錄活化蛋白;第一型干擾素誘發基因
出版社: 生物醫學研究所
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Type I interferon (IFN) is the first line of host defense against virus infection. Recent studies indicate the non-structural proteins of Japanese encephalitis virus (JEV) have antagonistic effects on type I IFN in vitro, but it is not consistent with the results in animal experiments and clinical cases. Therefore, we want to clarify whether blocking of type I IFN signaling by JEV is in a cell-specific manner. We found that the RNA expression of type I IFN was increased after JEV infection in microglia, but sightly affected in astrocytes. And its downstream signaling, transducers and activators of transcription (STAT) 1/2 protein family, was obviously phosphorylated in microglia. Astrocytes intrinsically expressed STAT1/2 in activated form, and not altered after JEV infection. Moreover, we also demonstrated similar results in protein expression of ISG15 and viperin, which are IFN-stimulated genes (ISGs).

On the other hand, the production of type I IFN in central nervous system (CNS) is little known. Glia cells are numerous in rat cerebral cortex and also important in immunomodulation. Hence, we obversed activation of mitogen-activated protein kinase (MAPK), increased transcription activity of nuclear factor-κB (NF-κB) after JEV infection in mixed glia. It is a pathway involved in the biosynthesis of type I IFN, and regulated by reactive oxygen species (ROS). Further analysis indicated that JEV-induced oxidative press is mainly from superoxide, which is produced through nicotinamide adenine dinucleotide phosphate (NADPH) oxidase in microglia. In addition, protein expression of inducible nitric oxide synthase (iNOS) and cyclooxygenase-2 (COX-2) were increased in microglia following JEV infection. Activated microglia, as evidenced by morphological transformation, play roles in neuroinflammation and may indirectly cause neuron death.

Molecular pathogenesis of JEV is still unclear, and there is no useful drug to cure Japanese encephalitis patients in clinical. In this study, we pointed out that glia cells produce type I IFN through ROS─MAPK─NF-κB pathway after JEV infection. And activated glia cells not only process inflammatory response, but also indirectly lead to neuron death. Importantly, the sensitivity to type I IFN of microglia and astrocytes seems to be different. Providing a platform for studying molecular pathogenesis of JEV, it is potential to develope another strategy against JEV depend on cell-specific virus susceptibility.

第一型干擾素是宿主細胞對抗病毒感染的第一道防線,近來研究指出日本腦炎病毒的非結構蛋白具有拮抗作用,但與活體動物實驗、臨床案例的結果並不一致。因此,我們想要釐清日本腦炎病毒拮抗第一型干擾素是否具有細胞特異性。本篇論文中,以日本腦炎病毒分別感染大鼠大腦皮質初代微神經膠細胞與星狀神經膠細胞,第一型干擾素 RNA 在前者有較為明顯的上升表現,而星狀神經膠細胞處理病毒後,除了本身第一型干擾素 RNA 的表現,僅微幅增加。第一型干擾素下游訊號路徑 signal transducers and activators of transcription (STAT)-1/2 蛋白家族,酪胺酸磷酸化位置在微神經膠細胞中明顯被活化,而星狀神經膠細胞本身就穩定表現活化態 STAT,沒有受到病毒感染而增加蛋白表現的幅度。進而探討下游第一型干擾素誘發基因,ISG15、viperin 抗病毒蛋白產物,也得到類似的結果。

此外,關於中樞神經系統如何產生第一型干擾素,知道的仍舊不多,而神經膠細胞除了參與免疫調節,也佔有大腦皮質大部分比例。因此,我們以日本腦炎病毒感染混合神經膠細胞後,發現 mitogen-activated protein kinase (MAPK) 路徑活化,促使 nuclear factor-κB (NF-κB) 進行轉錄生成第一型干擾素,且受到活性含氧物質的調控。進一步分析,觀察日本腦炎病毒造成的氧化壓力,主要來自微神經膠細胞,經由 nicotinamide adenine dinucleotide phosphate (NADPH) oxidase 產生超氧化物所致。先前實驗室研究已指出神經膠細胞參與發炎反應,間接導致神經元細胞死亡。在此,我們也觀察到病毒感染微神經膠細胞增加 inducible nitric oxide synthase (iNOS)、cyclooxygenase-2 (COX-2) 蛋白表現。

目前針對日本腦炎病患臨床上尚無有效治療方式,相關致病分子機轉的研究有待釐清。在本篇論文中,我們指出 ROS─MAPK─NF-κB 為一條參與神經膠細胞生成第一型干擾素的路徑,且神經膠細胞進行發炎反應間接導致神經元細胞死亡。另外,我們說明不同細胞對於第一型干擾素具有感受性的差異,導致抗病毒能力亦不相同,提供一個思考平台探討日本腦炎病毒致病分子機轉,期許發展出另一種策略對抗日本腦炎病毒。
其他識別: U0005-1411201110550200
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