本文選題：病毒侵染 + 主要穹窿蛋白�。� 參考：《武漢大學(xué)》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(gè)主要組成部分,它在細(xì)胞耐藥性、核質(zhì)運(yùn)輸、細(xì)胞信號(hào)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(cè)了HCV患者與健康人MVP mRNA和蛋白的表達(dá)差異。結(jié)果表明,和健康人比,HCV患者PBMC細(xì)胞中的MVP mRNA表達(dá)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(dá)水平也顯著升高。因?yàn)镠uh7細(xì)胞系和Huh7.5.1細(xì)胞系是一種有效的的HCV感染的細(xì)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(xì)胞系和Huh7.5.1細(xì)胞系,并檢測(cè)MVP表達(dá)水平的變化。結(jié)果表明,HCV JFH-1能誘導(dǎo)MVP表達(dá),并呈時(shí)間依賴和劑量依賴的方式。為探討HCV上調(diào)MVP表達(dá)水平的機(jī)理,我們通過PCR的方法獲得了MVP基因啟動(dòng)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(dòng)子的活力,進(jìn)一步研究表明,轉(zhuǎn)錄因子NF-κB和Sp1參與了NS5A對(duì)MVP的調(diào)控。 接著,我們探討了MVP對(duì)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(dá)和促進(jìn)蛋白的分泌,來抑制HCV的復(fù)制。進(jìn)一步研究發(fā)現(xiàn),過表達(dá)MVP能夠增加IRF7/NF-κB的表達(dá),并且促進(jìn)IRF7/NF-κB入核,但對(duì)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(dǎo)的IRF7/NF-κB的表達(dá)和入核,以及Ⅰ型干擾素mRNA的表達(dá)和蛋白的分泌。進(jìn)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(dǎo)MVP的表達(dá),而MVP反過來對(duì)這三種病毒都有抑制作用。小結(jié)：MVP是一種病毒誘導(dǎo)的宿主因子,它的表達(dá)能夠上調(diào)Ⅰ型干擾素,進(jìn)而對(duì)病毒產(chǎn)生抑制。 雖然對(duì)MVP在病毒復(fù)制和抗病毒反應(yīng)中的作用的許多細(xì)節(jié)還不是十分清楚,但是以上研究為揭示MVP的一個(gè)新功能(即細(xì)胞抵抗病毒作用)提供了新的證據(jù)。同時(shí)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(xué)位授予單位】：武漢大學(xué)
【學(xué)位級(jí)別】：博士
【學(xué)位授予年份】：2012
【分類號(hào)】：R392

【參考文獻(xiàn)】

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

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本文編號(hào)：2087981