本文選題：神經(jīng)管畸形　切入點(diǎn)：無腦畸形　出處：《中國協(xié)和醫(yī)科大學(xué)》2010年博士論文　論文類型：學(xué)位論文

【摘要】： 神經(jīng)管畸形(neural tube defects,NTDs)是由于在胚胎發(fā)育過程中,神經(jīng)管閉合不全所引起的一類先天畸形。NTDs屬多因素疾病,該病的發(fā)生主要是遺傳因素和環(huán)境因素及兩者相互作用的結(jié)果。這種畸形起因于神經(jīng)管閉合的失敗,所以影響神經(jīng)管發(fā)育的因素都可能促使其形成。MicroRNAs (miRNAs)是一類長約22個(gè)核苷酸的內(nèi)源性單鏈非編碼RNA,通過與靶mRNA完全或不完全互補(bǔ)配對(duì),導(dǎo)致靶mRNA降解或抑制其翻譯,從而在基因的表達(dá)調(diào)控中發(fā)揮著重要作用。目前研究表明miRNAs在神經(jīng)干細(xì)胞分化和哺乳動(dòng)物腦發(fā)育過程中起重要作用,提示miRNAs可能參與神經(jīng)管畸形的發(fā)生。但以往研究檢測的miRNA數(shù)量有限,并且國內(nèi)外尚無miRNA在胚胎腦組織中功能研究的報(bào)告。因此有必要深入開展miRNA在NTDs胚胎腦組織中的表達(dá)和功能研究,為深入了解NTDs的發(fā)病機(jī)理提供新的線索。本課題通過對(duì)于miRNAs在NTDs與正常胚胎腦組織的差異表達(dá)分析和特定miRNAs作用機(jī)制的闡釋,有助于我們進(jìn)一步提高對(duì)于NTDs發(fā)生發(fā)展的分子機(jī)制的認(rèn)識(shí)。 目的： 建立神經(jīng)管畸形與正常胚胎腦組織miRNAs的差異表達(dá)譜,利用生物信息學(xué)手段,預(yù)測差異表達(dá)顯著的miRNAs的靶基因,并進(jìn)一步分析靶基因的生物學(xué)功能及在神經(jīng)管畸形發(fā)生中的作用。為進(jìn)一步探討miRNAs在神經(jīng)管畸形發(fā)生機(jī)制中的作用提供依據(jù)。 方法： 利用1niRCURYTM LNA miRNA表達(dá)譜芯片分析無腦畸形殘余腦組織和正常胎兒腦組織miRNAs差異表達(dá)。利用NCodeTM qRT-PCR kit試劑盒,以U6 snRNA作為內(nèi)標(biāo),對(duì)差異表達(dá)顯著的miRNAs進(jìn)行驗(yàn)證。利用mirGEN v3數(shù)據(jù)庫預(yù)測差異表達(dá)顯著的9個(gè)miRNAs的靶基因,利用Human Protein Reference Database (HPRD)分析靶蛋白的相互作用,建立相互作用網(wǎng)。利用GeneCodis 2.0分析相互作用網(wǎng)所含蛋白質(zhì)的生物學(xué)功能,并以此推斷miRNAs在NTDs發(fā)生發(fā)展中的作用。 結(jié)果： 1. miRNA芯片檢測發(fā)現(xiàn)：實(shí)驗(yàn)組與對(duì)照相比,有213個(gè)miRNAs表達(dá)差異顯著(兩組差異2倍)。niRNAs表達(dá)上調(diào)2倍以上116個(gè)(其中包含1niRPLUS 9個(gè)),miRNAs表達(dá)下調(diào)2倍以上97個(gè)(其中包含miRPLUS 2個(gè)) 2.依據(jù)表達(dá)差異顯著性和在腦組織中表達(dá)的特異性,選擇9個(gè)miRNAs進(jìn)行real-time RT-PCR驗(yàn)證。miR-9、miR-212、miR-451、miR-198、miR-126、miR-124和miR-138檢測結(jié)果與miRNA芯片一致,miR-103/miR-107與miRNA芯片結(jié)果相反。表達(dá)上調(diào)：miR-451、miR-198和miR-126,表達(dá)下調(diào)：miR-9、miR-212、miR-103、miR-107、miR-124和miR-138。這9個(gè)miRNAs在NTDs和正常對(duì)照中的表達(dá)差異均具有統(tǒng)計(jì)學(xué)意義(p0.05) 3.經(jīng)過mirGEN v3數(shù)據(jù)庫分析,881個(gè)基因被預(yù)測為這9個(gè)1niRNAs的靶基因。其中79個(gè)基因包含在蛋白質(zhì)相互作用網(wǎng)中。 4.經(jīng)GeneCodis 2.0分析,這些蛋白參與基因轉(zhuǎn)錄、信號(hào)轉(zhuǎn)導(dǎo)、細(xì)胞周期、細(xì)胞生長、死亡和細(xì)胞間信號(hào)轉(zhuǎn)導(dǎo)這些重要的生物學(xué)過程；發(fā)揮蛋白結(jié)合、轉(zhuǎn)錄因子活化、受體結(jié)合及活化、核酸結(jié)合、DNA結(jié)合和轉(zhuǎn)移酶活化等生物學(xué)功能；并且它們涉及肌動(dòng)蛋白、Jak-STAT和MAPK等與神經(jīng)發(fā)育相關(guān)的信號(hào)通路。 結(jié)論： miRNAs在無腦畸形和正常對(duì)照胎兒腦組織中表達(dá)譜存在差異,可能參與了無腦畸形發(fā)生的分子機(jī)制。同時(shí),本研究繪制了靶基因的蛋白質(zhì)相互作用網(wǎng),并分析了這些靶蛋白的生物學(xué)功能,為深入研究這些miRNAs的調(diào)控機(jī)制及在NTDs發(fā)生發(fā)展中的作用提供了基礎(chǔ)。
[Abstract]:Neural tube defects (neural tube, defects, NTDs) is due to the process of embryonic development, a kind of neural tube malformation caused by the incomplete closure of.NTDs is a multifactorial disease, the incidence of the disease is genetic and environmental factors and the interaction between the two results. This deformity is attributed to the failure of neural tube closure, so factors affecting the development of the neural tube are likely to promote the formation of.MicroRNAs (miRNAs) is a single stranded endogenous 22 nucleotides encoding non RNA, with target mRNA complete or incomplete complementary, leading to degradation of target mRNA or inhibiting the translation, resulting in the regulation of gene expression plays an important role in the present. Research shows that miRNAs plays an important role in the process of development and differentiation of nerve stem cells in the mammalian brain, suggesting that miRNAs may be involved in the occurrence of neural tube defects. But previous studies detected a limited number of miRNA and And there is no report on miRNA function in embryonic brain tissue. So it is necessary to carry out the expression and function of miRNA in brain tissue in NTDs embryos, and provides new clues to the pathogenesis of in-depth understanding of NTDs. Through the research for the differences in miRNAs NTDs and normal embryonic brain tissue specific expression analysis and miRNAs the mechanism of interpretation, contribute to the improvement of our understanding of the molecular mechanism of the occurrence and development of NTDs.
Objective:
Differential expression of neural tube defects and normal embryonic brain tissue miRNAs established by the method of bioinformatics, gene expression of miRNAs was predicted differences, and further analysis of the biological function of target gene and tube malformation in nerve. In order to further explore the miRNAs incidence of neural tube defect mechanism provides the basis for.
Method錛，

本文編號(hào)：1639633