【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，屬于逆轉錄病毒。HIV-1長末重復端（long terminal repeat，LTR）上有一個調節(jié)基因tat編碼反式激活蛋白（trans-activator of transcription，Tat）。Tat在HIV病毒復制過程中是必需的，它可以調節(jié)病毒基因組的mRNA轉錄的起始，并與細胞的凋亡有關。Tat在HIV-1基因表達的調控和生命周期中發(fā)揮重要作用，使其成為抗HIV治療的新靶點。 本課題選擇對HIV-1Tat基因起調節(jié)作用的沉默信息調節(jié)子2類似物1——也叫人類去乙酰化酶1（silent mating type information regulation2homolog1，SIRT1）作為靶點，選取與之相關的尼克酰胺磷酸核糖轉移酶（nicotinamidephosphoribosyltransferase，Nampt）以及能與Nampt、SIRT1mRNA相作用的微小RNA（microRNAs）進行探索，檢驗是否在內源基因水平對Tat介導的HIV-1的轉錄與復制的調節(jié)起作用。 實驗結果：通過實驗發(fā)現(xiàn)一些microRNAs確實對Tat介導的HIV-1LTR的轉錄能夠起到有效的抑制作用。其中miR-34、miR-199a、miR-217都能夠通過對人類去乙酰化酶SIRT1的抑制作用對Tat介導的HIV-1的轉錄進行調控。與SIRT1緊密相關的尼克酰胺磷酸核糖轉移酶Nampt能通過調節(jié)NAD~+水平調控SIRT1的活性，Nampt與NAD~+水平緊密相關。由于SIRT1是NAD~+依賴的，我們選取可能影響Nampt的miR-182進行實驗，檢驗其對Tat介導的HIV-1轉錄的影響，實驗結果發(fā)現(xiàn)miR-182能有效的降低Nampt的蛋白表達，同時由于Nampt與SIRT1的正相關性，，miR-182能有效的降低SIRT1mRNA及蛋白水平的表達，這些microRNAs都能有效地增強下游核因子NF-κB的p65亞基的乙酰化表達，p65亞基乙酰化的增加會促使轉錄的活化促進HIV-1的轉錄。而這些microRNA的抑制劑則能有效降低乙�；痯65的表達，使得HIV-1轉錄過程受到阻礙。實驗表明通過尋找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.
【學位授予單位】：北京工業(yè)大學
【學位級別】：碩士
【學位授予年份】：2012
【分類號】：R373

【參考文獻】

相關期刊論文 前2條

1 孫曉娜;楊怡姝;曾毅;;反式激活應答元件RNA在HIV-1感染中的作用[J];中國生物化學與分子生物學報;2010年04期

2 張紅勝;周s

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