本文選題：病毒侵染 + 主要穹窿蛋白��； 參考：《武漢大學》2012年博士論文

【摘要】：丙型肝炎病毒(HCV)感染成為危害公眾健康的主要問題。全世界有超過1.7億人感染HCV,其中在中國有超過3000萬人感染HCV。主要穹窿蛋白(MVP)是穹窿蛋白的一個主要組成部分,它在細胞耐藥性、核質(zhì)運輸、細胞信號通路中起到關(guān)鍵作用。目前為止,關(guān)于HCV感染與MVP的關(guān)系尚不清楚。 為了探討HCV感染與MVP的關(guān)系,我們檢測了HCV患者與健康人MVP mRNA和蛋白的表達差異。結(jié)果表明,和健康人比,HCV患者PBMC細胞中的MVP mRNA表達水平明顯升高。同樣,在HCV患者血清和肝組織中,MVP蛋白的表達水平也顯著升高。因為Huh7細胞系和Huh7.5.1細胞系是一種有效的的HCV感染的細胞培養(yǎng)模型,所以我們使用HCV2a亞型復(fù)制子(JFH-1)侵染Huh7細胞系和Huh7.5.1細胞系,并檢測MVP表達水平的變化。結(jié)果表明,HCV JFH-1能誘導MVP表達,并呈時間依賴和劑量依賴的方式。為探討HCV上調(diào)MVP表達水平的機理,我們通過PCR的方法獲得了MVP基因啟動子,將它構(gòu)建到載體pGL3-basic上。研究發(fā)現(xiàn),HCVNS5A蛋白能夠上調(diào)MVP啟動子的活力,進一步研究表明,轉(zhuǎn)錄因子NF-κB和Sp1參與了NS5A對MVP的調(diào)控。 接著,我們探討了MVP對HCV的影響。結(jié)果表明,MVP能夠通過上調(diào)Ⅰ型干擾素：nRNA的表達和促進蛋白的分泌,來抑制HCV的復(fù)制。進一步研究發(fā)現(xiàn),過表達MVP能夠增加IRF7/NF-κB的表達,并且促進IRF7/NF-κB入核,但對IRF3沒有什么影響。將MVP干擾后,能夠抑制水皰性口炎病毒(VSV)誘導的IRF7/NF-κB的表達和入核,以及Ⅰ型干擾素mRNA的表達和蛋白的分泌。進一步研究發(fā)現(xiàn),水皰性口炎病毒(VSV)、A型流感病毒(IAV)、腸道病毒71(EV71)能誘導MVP的表達,而MVP反過來對這三種病毒都有抑制作用。小結(jié)：MVP是一種病毒誘導的宿主因子,它的表達能夠上調(diào)Ⅰ型干擾素,進而對病毒產(chǎn)生抑制。 雖然對MVP在病毒復(fù)制和抗病毒反應(yīng)中的作用的許多細節(jié)還不是十分清楚,但是以上研究為揭示MVP的一個新功能(即細胞抵抗病毒作用)提供了新的證據(jù)。同時為HCV的治療奠定理論基礎(chǔ)。
[Abstract]:Hepatitis C virus (HCV) infection has become a major public health problem. More than 170 million people worldwide are infected with HCV, of which more than 30 million are infected in China. Major fornix protein (MVP) is a major component of fornix protein, which plays a key role in cell resistance, nuclear and cytoplasmic transport, and cell signaling pathway. So far, the relationship between HCV infection and MVP is unclear. To investigate the relationship between HCV infection and MVP, we detected the difference of MVP mRNA and protein expression between HCV patients and healthy subjects. The results showed that the expression of MVP mRNA in PBMC cells of HCV patients was significantly higher than that in healthy subjects. Similarly, the expression of MVP protein in serum and liver tissues was significantly increased in HCV patients. Because Huh7 cell line and Huh7.5.1 cell line are an effective cell culture model of HCV infection, we used HCV2a subtype replicon (JFH-1) to infect Huh7 cell line and Huh7.5.1 cell line, and to detect the change of MVP expression level. The results showed that HCV JFH-1 could induce MVP expression in a time and dose dependent manner. In order to investigate the mechanism of HCV up-regulation of MVP expression, we obtained the promoter of MVP gene by PCR and constructed it into pGL3-basic vector. It was found that HCVNS5A protein could up-regulate the activity of MVP promoter. Further studies showed that NF- 魏 B and Sp1 were involved in the regulation of MVP by NS5A. Then, we studied the effect of MVP on HCV. The results showed that MVP could inhibit HCV replication by up-regulating the expression of type I interferon: nRNA and promoting protein secretion. Further studies showed that overexpression of MVP increased the expression of IRF7 / NF- 魏 B and enhanced the entry of IRF7 / NF- 魏 B into the nucleus, but had little effect on IRF3. The interference of MVP inhibited the expression and nucleation of IRF7 / NF- 魏 B induced by vesicular stomatitis virus (VSV), as well as the expression of interferon type I mRNA and the secretion of protein. Further studies showed that vesicular stomatitis virus (VSV) influenza A virus (IAV) and enterovirus 71 (EV71) could induce the expression of MVP and MVP could inhibit the expression of MVP. Conclusion: MVP is a virus-induced host factor, which can up-regulate the expression of interferon type I and thus inhibit the virus. Although many details of the role of MVP in viral replication and antiviral response are not well understood, the above studies provide new evidence to reveal a new function of MVP (cell resistance to virus). At the same time, lay a theoretical foundation for the treatment of HCV.
【學位授予單位】：武漢大學
【學位級別】：博士
【學位授予年份】：2012
【分類號】：R392

【參考文獻】

相關(guān)期刊論文 前3條

1 Susan L.Uprichard;;Hepatitis C Virus Experimental Model Systems and Antiviral drug Research[J];Virologica Sinica;2010年04期

2 Volker Meier;Sabine Mihm;Perdita Wietzke-Braun;Guliano Ramadori;;HCV-RNA positivity in peripheral blood mononuclear cells of patients with chronic HCV-infection: does it really mean viral replication?[J];World Journal of Gastroenterology;2001年02期

3 Miriam J Alter;;Epidemiology of hepatitis C virus infection[J];World Journal of Gastroenterology;2007年17期

，

本文編號：2087981