本文選題：結(jié)核病 + SNP��； 參考：《北京協(xié)和醫(yī)學院》2014年博士論文

【摘要】：結(jié)核病是嚴重危害人類健康的重要傳染病之一。過去二十年隨著現(xiàn)代結(jié)核病控制策略的普及,我國結(jié)核病防控工作取得了顯著成效。但是,作為全球二十二個結(jié)核病高負擔國家之一,我國的結(jié)核病疫情仍然非常嚴峻。面對這種形勢,尋找有效的方法控制結(jié)核病,降低結(jié)核病發(fā)病率和死亡率成為當務(wù)之急。然而,目前結(jié)核病診防治領(lǐng)域面臨著很多瓶頸問題：耐藥患者數(shù)量增多、新藥研發(fā)遲緩；診斷方法耗時且敏感性差,難以滿足臨床工作需要；唯一可選的預防性疫苗(卡介苗)對成年人保護效果不佳。新藥物、新診斷技術(shù)以及新疫苗的研發(fā)都離不開分子標識研究,尤其是具有我國自主知識產(chǎn)權(quán)的分子標識不僅僅是應(yīng)用研究的基礎(chǔ),也將為高危人群鑒定以及疾病發(fā)生發(fā)展的機制研究提供線索和證據(jù)。本研究旨在通過分子流行病學研究方法,采用病例-對照的研究設(shè)計,分析宿主單核苷酸多態(tài)性(Single Nucleotide Polymorphism, SNP)及血漿microRNA (miRNA)與結(jié)核分枝桿菌感染狀態(tài)及結(jié)核病發(fā)生之間的相關(guān)性,探討結(jié)核分枝桿菌感染到結(jié)核病發(fā)生的進程中有潛在應(yīng)用價值的宿主分子標識。 結(jié)核病發(fā)病相關(guān)宿主SNP的病例-對照研究納入三組調(diào)查對象(涂陽活動性肺結(jié)核患者組,結(jié)核分枝桿菌潛伏感染組和感染陰性對照組)共計600例,應(yīng)用Illumina公司的Goldengate芯片對44個候選SNP進行檢測。研究結(jié)果發(fā)現(xiàn),IFNG2109G/A位點GG基因型與結(jié)核病發(fā)病風險的降低顯著相關(guān)(OR=0.54,95%CI:0.30-0.98); TLR91174A/G位點GG基因型(OR=1.87,95%CI:1.08-3.23),及TLR91635位點AA基因型(OR=1.90,95%CI:1.09-3.31)均與較高的結(jié)核病發(fā)病風險顯著相關(guān)。同時,TLR91174G/G和IFNG2109A/A基因型表現(xiàn)出協(xié)同效應(yīng)。其余候選SNP在本研究中未表現(xiàn)出與結(jié)核分枝桿菌感染或者結(jié)核病發(fā)病的顯著相關(guān)性。以上結(jié)果提示在中國人群中TLR9和IFN-γ的基因多態(tài)性可能與肺結(jié)核的發(fā)病風險相關(guān),為進一步探討TLR9/IFN-γ通路在從結(jié)核分枝桿菌感染到致病的過程中所發(fā)揮的調(diào)控作用提供了思路。 結(jié)核病發(fā)病與血漿miRNA表達水平的相關(guān)性研究納入三組調(diào)查對象(涂陽活動性肺結(jié)核患者組,結(jié)核分枝桿菌潛伏感染組和感染陰性對照組)共計34例。針對調(diào)查對象的血漿樣本,應(yīng)用Agilent human miRNA芯片對1887個人類miRNA的表達水平進行檢測,隨后對篩選得到的11個差異表達miRNA應(yīng)用Real-time qPCR (Fold Change4, P0.01)方法進行驗證,最后對通過驗證的差異表達miRNA進行靶基因預測和調(diào)控網(wǎng)絡(luò)構(gòu)建。研究結(jié)果發(fā)現(xiàn),在與對照組(潛伏感染組和感染陰性對照組)的比較中,hsa-let-7b-5p和hsa-miR-30b-5p在結(jié)核病患者血漿中的表達水平顯著上調(diào)(P0.05),基于統(tǒng)計模型的靶基因調(diào)控網(wǎng)絡(luò)預測結(jié)果進一步提示,這兩個miRNA可能通過調(diào)控炎癥因子、細胞凋亡因子等的表達而參與結(jié)核病的發(fā)生。該研究結(jié)果為深入研究血漿miRNA在結(jié)核病發(fā)病過程中的調(diào)控作用及其作為診斷分子標識的應(yīng)用價值提供了線索,也為進一步的大樣本驗證以及分子標識的優(yōu)化組合提供了科學依據(jù)。
[Abstract]:Tuberculosis is one of the important infectious diseases seriously endangering human health. With the popularization of modern tuberculosis control strategy in the past twenty years, the tuberculosis prevention and control work has achieved remarkable results. However, as one of the twenty-two countries with high tuberculosis burden in the world, the epidemic situation in China is still very severe. Effective methods to control tuberculosis and reduce the incidence and mortality of tuberculosis have become the top priority. However, there are many bottlenecks in the field of tuberculosis treatment and prevention: the number of drug resistant patients, the slow development of new drugs, the time-consuming and poor sensitivity of the diagnostic methods, and the difficulty of meeting the needs of clinical work; the only optional preventive vaccine ( Bacillus Calmette Guerin (BCG) is not effective in protecting adults. New drugs, new diagnostic techniques and new vaccine research and development can not be separated from molecular identification research. Especially, molecular markers with independent intellectual property rights in China are not only the basis of application research, but also provide clues and evidence for the study of high-risk population and the mechanism of disease and development. The purpose of this study is to analyze the correlation between the Single Nucleotide Polymorphism (SNP) and the plasma microRNA (miRNA) and the infection status of Mycobacterium tuberculosis and the occurrence of tuberculosis through a case control study. The purpose of this study is to explore the infection of Mycobacterium tuberculosis to tuberculosis. There are potentially valuable host molecular markers in the process of being born.
A case-control study of the host SNP of tuberculosis associated with three groups of subjects (smear positive pulmonary tuberculosis patients, Mycobacterium tuberculosis latent infection group and negative control group) totaled 600 cases, and 44 candidate SNP were tested with the Goldengate chip of Illumina company. The results of the study found that the IFNG2109G/A locus GG genotypes There was a significant correlation with the reduction of the risk of tuberculosis (OR=0.54,95%CI:0.30-0.98); the TLR91174A/G locus GG genotype (OR=1.87,95%CI:1.08-3.23) and the TLR91635 locus AA genotype (OR=1.90,95%CI:1.09-3.31) were significantly related to the higher risk of tuberculosis. At the same time, the co effect of the TLR91174G/G and IFNG2109A/A genotypes showed a synergistic effect. The candidate SNP did not show a significant correlation with Mycobacterium tuberculosis or tuberculosis in this study. The above results suggest that the genetic polymorphism of TLR9 and IFN- gamma may be associated with the risk of tuberculosis in Chinese people, so as to further explore the process of TLR9/IFN- gamma pathway in the process of infection from Mycobacterium tuberculosis to disease. It provides a way of thinking to play the role of regulation and control.
The correlation of tuberculosis incidence and plasma miRNA expression level was included in three groups of subjects (smear positive pulmonary tuberculosis patients, Mycobacterium tuberculosis latent infection group and negative control group) in total 34 cases. The expression level of 1887 human miRNA was examined by Agilent human miRNA chip. Then, the selected 11 differential expression miRNA applications, Real-time qPCR (Fold Change4, P0.01), were verified. Finally, the target gene prediction and regulation network were constructed by the differential expression of the verified miRNA. The results were found in the comparison with the control group (latent infection group and infection negative control group), hsa-let-7b-5p and The expression level of hsa-miR-30b-5p in the plasma of the patients with tuberculosis was significantly up-regulated (P0.05). The prediction results of the target gene regulation network based on the statistical model suggest that these two miRNA may be involved in the development of tuberculosis by regulating the expression of inflammatory factors and apoptosis factors. The result of this study is to study the plasma miRNA in tuberculosis. The regulatory role in the pathogenesis of the disease and its application value as a diagnostic molecular marker provides a clue, and provides a scientific basis for further large sample validation and the optimization of molecular markers.

【學位授予單位】：北京協(xié)和醫(yī)學院
【學位級別】：博士
【學位授予年份】：2014
【分類號】：R52

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相關(guān)博士學位論文 前1條

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本文編號：1845717