脂滴和脂代谢在肠道病毒71型增殖过程中的作用研究

发布时间：2018-07-06 10:35

本文选题：肠道病毒71型 + EV71　；参考：《南京大学》2014年硕士论文

【摘要】：肠道病毒71型(enterovirus71, EV71)是单股正链无包膜RNA病毒,属于小核糖核酸病毒科、肠道病毒属,是引起婴幼儿手足口病(hand-foot and mouth disease; HFMD)的主要病原体之一。临床上EV71可致婴幼儿手足口病并诱发严重的神经系统并发症,目前其致病机制仍不清楚。大量的研究表明EV71病毒在被感染的细胞胞浆内复制核酸并合成蛋白,然而在细胞胞浆中复制的具体机制及复制的亚细胞位点却没有相关的研究报道。本研究初步证明EV71的复制是在细胞胞浆中进行,并且观察到病毒复制过程中所形成的双链RNA(double-stranded RNA, dsRNA)中间体是存在于细胞胞浆中特征性的颗粒结构。随后进一步的研究发现病毒复制过程中所形成的dsRNA与细胞胞浆中的脂滴(lipid droplet, LD)以及围脂滴蛋白(perilipin A)可以共定位,并且随着感染时间的延长这种共定位颗粒会逐渐增大；在细胞感染前4h应用脂滴形成的阻断剂脂酰-CoA合成酶特异性抑制物(Triacsin C)、脂肪酸合成酶抑制物(C75)及脂肪分化相关蛋白特异性抑制物(TOFA)进行药物预处理后我们观察到被感染的细胞胞浆中,这种共定位颗粒在不同时间点颗粒数明显减少,并且不随感染时间的延长而增大；同时,感染后细胞上清病毒滴度与对照相比明显减低。本次研究表明。特异性抑制长链脂肪酸合成可以有效地抑制脂滴的形成,从而抑制EV71的复制增殖。证明EV71病毒复制与细胞脂质代谢相关,说明了EV71感染过程中病毒复制的可能机制,为新一代抗EV71病毒药物的研究提供新的思路。
[Abstract]:Enterovirus71 (EV71) is a single-strand positive strand non-encapsulated RNA virus, belonging to the small RNA virus family. Enterovirus 71 is one of the main pathogens of hand, foot and mouth disease (hand-foot and mouth disease;) in infants. EV71 can cause hand, foot and mouth disease in infants and induce severe neurological complications. The pathogenesis of EV71 is still unclear. A large number of studies have shown that EV71 virus replicates nucleic acids and synthesized proteins in the cytoplasm of infected cells. However, there are no related studies on the mechanism and subcellular sites of replication in the cytoplasm of EV71 virus. In this study, we preliminarily demonstrated that the replication of EV71 was carried out in the cytoplasm of the cells, and it was observed that the intermediate of double-stranded RNA (dsRNA), which was formed during the replication of the virus, was a characteristic granular structure in the cytoplasm of the cells. Then further studies showed that dsRNAs formed during viral replication could be co-located with lipid droplet (lipid droplet, LD in cytoplasm and peri lipid drop protein (perilipin A), and the co-localization particles would increase gradually with the prolongation of infection time. After pretreatment with lipid droplet blockers (triacsin C), fatty acid synthase inhibitors (C75) and adipose differentiation related protein specific inhibitors (TOFA) 4 hours before infection, we observed that In the cytoplasm of infected cells, The number of co-localized particles decreased significantly at different time points and did not increase with the prolongation of infection time, and the virus titer of supernatant of the infected cells decreased significantly compared with the control. This study shows that. Specific inhibition of long chain fatty acid synthesis can effectively inhibit the formation of lipid droplets and thus inhibit the replication and proliferation of EV71. It is proved that the replication of EV71 virus is related to the lipid metabolism of cells, and the possible mechanism of virus replication during EV71 infection is explained, which provides a new idea for the study of a new generation of anti-EV71 drugs.
【学位授予单位】：南京大学
【学位级别】：硕士
【学位授予年份】：2014
【分类号】：R725.1

【相似文献】