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標題: | 應用三段四極柱串聯式質譜儀分析受茲卡病毒感染之白線斑蚊其含磷酸膽鹼脂質組成研究 Using UPLC-QqQ-MS/MS based lipidomic approach to analyze phosphorylcholine-containing lipid compositions in Zika virus-infected Aedes albopictus |
作者: | Ya-Chih Cheng 鄭雅之 |
指導教授: | 蔡坤憲(Kun-Hsien Tsai) |
關鍵字: | 脂質體學,茲卡病毒,白線斑蚊,磷脂醯膽鹼,鞘磷脂, Lipidomics,Zika virus,Ae. albopictus,Phosphatidylcholine,Sphingomyelin, |
出版年 : | 2020 |
學位: | 碩士 |
摘要: | 白線斑蚊(Aedes albopictus)是全球重要的病媒之一,可傳播茲卡病毒感染症等蟲媒傳染病。這些病毒在感染細胞、細胞中複製和離開細胞時,都需要宿主提供脂質製膜狀構造。而脂質的結構、功能和脂質與病毒間的關係,可藉由脂質體學的方法來了解。先前的研究發現,部分RNA病毒與含磷酸膽鹼(phosphorylcholine)的脂質如磷脂醯膽鹼(phosphatidylcholine,PC)和鞘磷脂(sphingomyelin,SM)有關,並指出特定脂質的表現有助於病毒的複製和包裝。但由於多數的研究將焦點放在病毒與宿主間的關係,而非病毒與病媒間關係,使得相關研究較少。因此,本研究的目的是應用極致液相層析儀三段四極柱串聯式質譜儀,來了解受茲卡病毒亞洲型和非洲型感染的病媒細胞C6/36和經中胸注射的白線斑蚊雌蚊其頭胸部和腹部中的感染情況差異,與其含磷酸膽鹼脂質組成變化間的關係。 在蚊蟲細胞層級和蚊蟲個體層級的研究結果顯示在亞洲型和非洲型茲卡病毒的感染下,PCs和SMs相較於控制組有顯著表現量差異。首先,在兩者都觀察到PCs的表現量有顯著上升。另一方面,Lyso-PCs在受感染的細胞中顯著上升,但在蚊蟲腹部中卻呈下降趨勢。而SMs表現量變化則僅在細胞層級中觀察到,但在亞洲型與非洲型病毒的感染下有顯著差異。Lyso-PCs表現量的上升,與SMs表現量的下降,可能與非洲株病毒的感染速度和毒性有關。然而,在蚊蟲腹部中的僅發現同樣有上升變化的PCs,在Lyso-PCs和SMs中的發現與細胞層級有所不同,強調在蚊蟲個體和組織中進行實驗以反映脂質變化真實性的重要。因此,此研究描述了茲卡病毒亞洲型和非洲型在感染病媒細胞層級和個體層級的變化情形,並可增進對於茲卡病毒在病媒中感染、複製與脂質間關係更進一步的了解。 Aedes albopictus is an important vector that can transmit vector-borne diseases such as Zika virus (ZIKV) disease. The infection of these arbovirus depends on hosts’ lipid-made membranes. Recent studies that applied lipidomic approaches demonstrated the perturbation of host phosphorylcholine-containing lipids like phosphatidylcholines (PCs) and sphingomyelins (SMs) at viral replication sites and packing vesicles that aid the formation of viral replication complexes (VRCs) and increase membrane fluidity. However, the understanding about ZIKV infection from the aspects of vectors are not well developed. This study aimed to provide information on the relationships between Asian strain and African strain of ZIKV infection and phosphorylcholine-containing lipid composition in the vector by applying UPLC-QqQ-MS/MS in order to compare the virulence and the expression of PCs and SMs in cell and individual levels. The results were acquired from the extracted lipid of ZIKV infected C6/36 cells and intrathoracically infected Aedes albopictus mosquitoes. And it showed that in both cell and individual levels, the lipid composition was significantly different between ZIKV Asian and African strains-infected ones and uninfected control. First, in infected C6/36 cells and mosquito bodies, PCs were observed to increase. On the other hand, the changing tendencies of Lyso-PCs were increasing in infected C6/36 cells but decreasing in mosquito bodies. As for SMs, the different expression levels were only observed in ZIKV Asian and African strain-infected C6/36 cells which could be related to the different virulence and the infection stage. The increasing levels of Lyso-PCs and the decreasing levels of SMs can explain the more rapid and virulent properties of ZIKV African strain. And the different expression of lipids between cell and individual levels indicated the complex environment in the whole mosquito, which emphasized on the importance of in vivo experiments. These novel findings can provide basic biological information for understanding the relationship between ZIKV infection, replication and lipid composition in mosquito vector. And provide insights of the lipid composition from the aspect of vectors. |
URI: | http://tdr.lib.ntu.edu.tw/jspui/handle/123456789/73082 |
DOI: | 10.6342/NTU202004460 |
全文授權: | 有償授權 |
顯示於系所單位: | 環境與職業健康科學研究所 |
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