【摘要】：人類免疫缺陷病毒（human immunodeficiency virus，HIV）是獲得性免疫缺陷綜合癥（acquired immune deficiency syndrome，AIDS）的病原體，屬于逆轉(zhuǎn)錄病毒。HIV-1長(zhǎng)末重復(fù)端（long terminal repeat，LTR）上有一個(gè)調(diào)節(jié)基因tat編碼反式激活蛋白（trans-activator of transcription，Tat）。Tat在HIV病毒復(fù)制過程中是必需的，它可以調(diào)節(jié)病毒基因組的mRNA轉(zhuǎn)錄的起始，并與細(xì)胞的凋亡有關(guān)。Tat在HIV-1基因表達(dá)的調(diào)控和生命周期中發(fā)揮重要作用，使其成為抗HIV治療的新靶點(diǎn)。 本課題選擇對(duì)HIV-1Tat基因起調(diào)節(jié)作用的沉默信息調(diào)節(jié)子2類似物1——也叫人類去乙�；�1（silent mating type information regulation2homolog1，SIRT1）作為靶點(diǎn)，選取與之相關(guān)的尼克酰胺磷酸核糖轉(zhuǎn)移酶（nicotinamidephosphoribosyltransferase，Nampt）以及能與Nampt、SIRT1mRNA相作用的微小RNA（microRNAs）進(jìn)行探索，檢驗(yàn)是否在內(nèi)源基因水平對(duì)Tat介導(dǎo)的HIV-1的轉(zhuǎn)錄與復(fù)制的調(diào)節(jié)起作用。 實(shí)驗(yàn)結(jié)果：通過實(shí)驗(yàn)發(fā)現(xiàn)一些microRNAs確實(shí)對(duì)Tat介導(dǎo)的HIV-1LTR的轉(zhuǎn)錄能夠起到有效的抑制作用。其中miR-34、miR-199a、miR-217都能夠通過對(duì)人類去乙�；窼IRT1的抑制作用對(duì)Tat介導(dǎo)的HIV-1的轉(zhuǎn)錄進(jìn)行調(diào)控。與SIRT1緊密相關(guān)的尼克酰胺磷酸核糖轉(zhuǎn)移酶Nampt能通過調(diào)節(jié)NAD~+水平調(diào)控SIRT1的活性，Nampt與NAD~+水平緊密相關(guān)。由于SIRT1是NAD~+依賴的，我們選取可能影響Nampt的miR-182進(jìn)行實(shí)驗(yàn)，檢驗(yàn)其對(duì)Tat介導(dǎo)的HIV-1轉(zhuǎn)錄的影響，實(shí)驗(yàn)結(jié)果發(fā)現(xiàn)miR-182能有效的降低Nampt的蛋白表達(dá)，同時(shí)由于Nampt與SIRT1的正相關(guān)性，，miR-182能有效的降低SIRT1mRNA及蛋白水平的表達(dá)，這些microRNAs都能有效地增強(qiáng)下游核因子NF-κB的p65亞基的乙�；磉_(dá)，p65亞基乙酰化的增加會(huì)促使轉(zhuǎn)錄的活化促進(jìn)HIV-1的轉(zhuǎn)錄。而這些microRNA的抑制劑則能有效降低乙�；痯65的表達(dá)，使得HIV-1轉(zhuǎn)錄過程受到阻礙。實(shí)驗(yàn)表明通過尋找microRNAs作為抗HIV-1的新型治療方式是可行的。
[Abstract]:Human immunodeficiency virus (human immunodeficiency virus,HIV) is the pathogen of acquired immunodeficiency syndrome (acquired immune deficiency syndrome,AIDS) and belongs to retrovirus. HIV-1 long terminal repeat (long terminal repeat, There is a regulatory gene tat encoding transactivator protein (trans-activator of transcription,Tat). Tat) on LTR that is necessary in the replication of the HIV virus, which regulates the initiation of mRNA transcription in the viral genome. Tat plays an important role in the regulation and life cycle of HIV-1 gene expression, making it a new target of anti-HIV therapy. In this study, we selected analogue 1 of silencing information regulator 2, also known as human deacetylase 1 (silent mating type information regulation2homolog1,SIRT1, which regulates the HIV-1Tat gene, as the target, and selected the related nicotinamidephosphoribosyltransferase, ribonucleosyltransferase (nicotinamidephosphoribosyltransferase,). Nampt) and tiny RNA (microRNAs) that interact with Nampt,SIRT1mRNA were explored to examine whether Tat mediated HIV-1 transcription and replication were regulated at the endogenous gene level. The results showed that some microRNAs could inhibit the transcription of HIV-1LTR mediated by Tat. MiR-34,miR-199a,miR-217 can regulate the transcription of HIV-1 mediated by Tat through the inhibition of human deacetylase SIRT1. Nicotinamide phosphate ribonucleosyltransferase (Nampt), closely related to SIRT1, can regulate the activity of SIRT1 by regulating the level of NAD~, and Nampt is closely related to the level of NAD~. Since SIRT1 is NAD~ dependent, we selected miR-182 that may affect Nampt to test its effect on HIV-1 transcription mediated by Tat. The results showed that miR-182 could effectively reduce the expression of Nampt protein. At the same time, because of the positive correlation between Nampt and SIRT1, miR-182 can effectively reduce the expression of SIRT1mRNA and protein. These microRNAs can effectively enhance the acetylation of p65 subunit of downstream nuclear factor NF- 魏 B. An increase in acetylation of p65 subunits promotes the activation of transcription to promote the transcription of HIV-1. These inhibitors of microRNA can effectively reduce the expression of acetylated p65 and block the process of HIV-1 transcription. The experiment shows that it is feasible to find microRNAs as a new type of anti-HIV-1 therapy.
【學(xué)位授予單位】：北京工業(yè)大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2012
【分類號(hào)】：R373

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相關(guān)期刊論文 前2條

1 孫曉娜;楊怡姝;曾毅;;反式激活應(yīng)答元件RNA在HIV-1感染中的作用[J];中國(guó)生物化學(xué)與分子生物學(xué)報(bào);2010年04期

2 張紅勝;周s

本文編號(hào)：2298587