汉坦病毒体外诱导HSP70表达的定量研究

发布时间：2018-06-06 21:44

本文选题：汉坦病毒 + 热休克蛋白70　；参考：《第四军医大学》2005年硕士论文

【摘要】：热休克反应(Heat-Shock Response,HSR)是原核生物和真核生物受到不利或有害外界因素刺激后发生的一种生理性的快速短暂的细胞代谢调节,是一种细胞自身保护性反应。该过程以选择性上调表达热休克蛋白(Heat-Shock Proteins,HSPs)为特点。研究表明,HSP70作为一类“分子伴侣”(molecular chaperones),在生理状态下,参与多种蛋白质正常的折叠、装配及转运;在应激情况下可限制变性或损伤蛋白质的聚集从而抗细胞损伤,是细胞生存所必需的重要蛋白质。汉坦病毒(Hantavirus,HTV)是引起肾综合征出血热(Hemorrhagic Fever with Renal Syndrome,HFRS)的病原体,本课题组在前期研究发现,HFRS病人多种组织中有HSP70的表达,与HTV抗原存在共存和互相抑制关系,提示HTV感染是参与诱导HSR的重要应激因素之一。但是,目前对于HTV体外诱导细胞HSP70表达的规律及意义知之甚少。本课题在前期研究基础上,进一步观察了HTV感染诱导Vero-E6细胞和人脐静脉血管内皮细胞(Human Umbilical Vein Vascular Endothelial Cell,HUVEC)的热休克反应,定量分析HSP70的表达规律并探讨其意义: 1.Vero-E6细胞感染HTV后72 hr内不同时间点HSP70的表达规律:实验性体外感染Hantaan 76-118,免疫细胞化学染色证实HTV可直接感染Vero-E6细胞。用原位杂交和RT-PCR方法检测发现,在感染后0.5 hrHSP70 mRNA即出现升高,感染后12 hr达到高峰,并持续高表达至72 hr。用免疫细胞化学染色和免疫荧光检测发现,感染后2 hr可检测到HSP70蛋白表达增加,感染后12 hr达到高峰,并持续高表达至72 hr。 2.实验发现,HTV体外感染可以直接感染HUVEC并诱导热休克反应表达HSP70,与Vero-E6细胞有相似的表达规律。用原位杂交和RT-PCR
[Abstract]:Heat shock response Heat-Shock response (HSRs) is a physiological, rapid and transient regulation of cell metabolism after prokaryotes and eukaryotes are stimulated by adverse or harmful external factors. Studies have shown that HSP70, as a class of molecular chaperones, is involved in the normal folding, assembly and transport of many proteins under physiological conditions. It is an important protein necessary for cell survival. Hantavirus hantavirus (HTV) is the pathogen causing hemorrhagic fever with renal syndrome (HFRS). Our previous study found that HSP70 was expressed in various tissues of patients with HFRS, and there was a coexisting and mutual inhibition relationship between HSP70 and HTV antigen. It is suggested that HTV infection is one of the important stress factors involved in HSR induction. However, little is known about the regulation and significance of HSP70 expression induced by HTV in vitro. Based on previous studies, the heat shock responses of Vero-E6 cells and human umbilical vein endothelial cells (HUVECs) induced by HTV infection were further studied. Quantitative analysis of HSP70 expression and its significance: 1. Expression of HSP70 in Vero-E6 cells at different time points within 72 hours after HTV infection: Hantaan 76-118 was infected in vitro. Immunocytochemical staining confirmed that HTV could directly infect Vero-E6 cells. By in situ hybridization and RT-PCR, it was found that the mRNA of HSP70 increased at 0.5 h after infection, reached the peak at 12 h after infection, and continued to be overexpressed to 72 hr. Immunocytochemical staining and immunofluorescence assay showed that the expression of HSP70 protein increased at 2 hours after infection, reached the peak at 12 hours after infection, and continued to be high to 72 hr. 2. It was found that HTV infection in vitro could directly infect HUVEC and induce heat shock response to express HSP70, which was similar to that of Vero-E6 cells. In situ hybridization and RT-PCR
【学位授予单位】：第四军医大学
【学位级别】：硕士
【学位授予年份】：2005
【分类号】：R373

【共引文献】