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標題: | 剖析登革病毒蛋白酵素造成登革出血以及微血管內皮細胞凋亡之分子機制 To dissect the mechanism of dengue viral protease-mediated hemorrhage and endothelial cell apoptosis |
作者: | Jung-Chen Lin 林榮辰 |
指導教授: | 伍安怡(Betty A. Wu-Hsieh) |
關鍵字: | 登革出血致病機制,登革蛋白酵素,IκB蛋白,NF-κB,血管內皮細胞死亡, Pathogenesis of dengue haemorrhage,Dengue viral protease,IκB proteins,NF-κB,Endothelial cell death, |
出版年 : | 2014 |
學位: | 博士 |
摘要: | 登革出血現象經常伴隨著一系列登革病症而發生。許多研究集中於探討免疫因子在登革出血致病過程中的角色,對於登革病毒本身如何參與在登革疾病過程卻很少探討。登革病毒抗原會存在具有嚴重登革症狀的病人之血管內皮細胞中,而血管內皮細胞的凋亡現象也能在病人身上以及動物疾病模式中被觀察到。這些結果顯示血管內皮細胞凋亡與疾病發展過程之間存在著關連性。
本實驗研究目的為探討登革病毒對於血管內皮細胞凋亡以及登革出血現象的分子致病機制。先前動物實驗結果顯示血管內皮細胞的凋亡對於登革出血發展扮演重要的角色。我們的研究結果顯示在血管內皮細胞凋亡的過程中,登革病毒蛋白酵素是一個致病因子。並且鑑定出登革病毒蛋白酵素會與宿主蛋白IκBα和IκBβ結合。登革病毒蛋白酵素透過切割IκBα和IκBβ與活化IKK 進而活化NF-κB而導致血管內皮細胞凋亡。利用重組腺相關病毒之活體內的實驗系統,我們將登革病毒蛋白酵素送入老鼠皮膚血管內皮細胞,發現登革病毒蛋白酵素會造成老鼠皮下出血。此外,大量表現IκBα和IκBβ蛋白會減低血管內皮細胞凋亡的發生。實驗結果也發現登革蛋白酵素能和登革病毒感染一樣造成血管內皮細胞中NF-κB 活化,並造成IL-32 與TNFRSF9 表現量增加。IL-32 與TNFRSF9 均被報導過在發炎過程中扮演著重要角色。 本篇研究結論為登革病毒造成的細胞凋亡與登革出血現象是經由登革病毒蛋白酵素引發NF-κB活化來達成。這些發現對於我們在了解登革病毒如何參與血管內皮細胞凋亡以及出血過程的分子機制,提供了一個新的方向。 Hemorrhagic manifestations occur frequently accompanying a wide range of dengue disease syndromes. Much work has focused on the contribution of immune factors to the pathogenesis of hemorrhage, but how dengue virus (DENV) participates in the pathogenic process has never been explored. DENV antigen is detected in the endothelium of tissues from patients with severe disease. The link between endothelial cell death and severe disease is reported in fatal cases and in dengue hemorrhage mouse model. The goal of this study is to explore the molecular basis of the contribution of DENV to endothelial cell death and hemorrhage development. Our previous results showed in dengue hemorrhage mouse model that endothelial cell apoptosis is important to hemorrhage development in mice. We discovered that DENV protease is a viral factor that participates in the pathogenesis of dengue hemorrhage and interacts with new cellular proteins, IκBα and IκBβ. DENV protease activates NF-κB through cleavage of IκBα and IκBβ and activation of IKK, thereby inducing endothelial cell apoptosis. In addition, DENV protease targeted to endothelium by AAV9 induces hemorrhage in mice. Over-expression of IκBα or IκBβ protects endothelial cells from DENV-induced apoptosis. Transduction of endothelial cells with DENV protease, as it was in DENV infection, induces NF-κB activation and in the mean time up-regulated IL-32 and TNFRSF9, both of which are known to play a role in inflammatory response. In summary, these results reveal DENV induces endothelial cell apoptosis and hemorrhage development through viral protease-induced NF-κB activation. Our findings provide novel insights into the molecular mechanism of how DENV causes endothelial cell apoptosis and participate in the hemorrhage development. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/56046 |
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顯示於系所單位: | 免疫學研究所 |
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