本文選題：漢坦病毒 + 熱休克蛋白70�。� 參考：《第四軍醫(yī)大學(xué)》2005年碩士論文

【摘要】：熱休克反應(yīng)(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發(fā)生的一種生理性的快速短暫的細(xì)胞代謝調(diào)節(jié),是一種細(xì)胞自身保護(hù)性反應(yīng)。該過(guò)程以選擇性上調(diào)表達(dá)熱休克蛋白(Heat-Shock Proteins,HSPs)為特點(diǎn)。研究表明,HSP70作為一類“分子伴侶”(molecular chaperones),在生理狀態(tài)下,參與多種蛋白質(zhì)正常的折疊、裝配及轉(zhuǎn)運(yùn);在應(yīng)激情況下可限制變性或損傷蛋白質(zhì)的聚集從而抗細(xì)胞損傷,是細(xì)胞生存所必需的重要蛋白質(zhì)。漢坦病毒(Hantavirus,HTV)是引起腎綜合征出血熱(Hemorrhagic Fever with Renal Syndrome,HFRS)的病原體,本課題組在前期研究發(fā)現(xiàn),HFRS病人多種組織中有HSP70的表達(dá),與HTV抗原存在共存和互相抑制關(guān)系,提示HTV感染是參與誘導(dǎo)HSR的重要應(yīng)激因素之一。但是,目前對(duì)于HTV體外誘導(dǎo)細(xì)胞HSP70表達(dá)的規(guī)律及意義知之甚少。本課題在前期研究基礎(chǔ)上,進(jìn)一步觀察了HTV感染誘導(dǎo)Vero-E6細(xì)胞和人臍靜脈血管內(nèi)皮細(xì)胞(Human Umbilical Vein Vascular Endothelial Cell,HUVEC)的熱休克反應(yīng),定量分析HSP70的表達(dá)規(guī)律并探討其意義: 1.Vero-E6細(xì)胞感染HTV后72 hr內(nèi)不同時(shí)間點(diǎn)HSP70的表達(dá)規(guī)律:實(shí)驗(yàn)性體外感染Hantaan 76-118,免疫細(xì)胞化學(xué)染色證實(shí)HTV可直接感染Vero-E6細(xì)胞。用原位雜交和RT-PCR方法檢測(cè)發(fā)現(xiàn),在感染后0.5 hrHSP70 mRNA即出現(xiàn)升高,感染后12 hr達(dá)到高峰,并持續(xù)高表達(dá)至72 hr。用免疫細(xì)胞化學(xué)染色和免疫熒光檢測(cè)發(fā)現(xiàn),感染后2 hr可檢測(cè)到HSP70蛋白表達(dá)增加,感染后12 hr達(dá)到高峰,并持續(xù)高表達(dá)至72 hr。 2.實(shí)驗(yàn)發(fā)現(xiàn),HTV體外感染可以直接感染HUVEC并誘導(dǎo)熱休克反應(yīng)表達(dá)HSP70,與Vero-E6細(xì)胞有相似的表達(dá)規(guī)律。用原位雜交和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
【學(xué)位授予單位】：第四軍醫(yī)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2005
【分類號(hào)】：R373

【共引文獻(xiàn)】

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本文編號(hào)：1988214