本文選題：配子-胚胎源性疾病 + iTRAQ��； 參考：《浙江大學(xué)》2016年博士論文

【摘要】：第一部分基于于iTRAQ-2D-LC-MS/MS的子癇前期子代臍動(dòng)脈血管的蛋白質(zhì)組學(xué)分析流行病學(xué)調(diào)查顯示子癇前期對母親和胎兒的健康都有重大影響,宮內(nèi)的不良環(huán)境可能導(dǎo)致成人心血管疾病的發(fā)生,但對其分子機(jī)制知之甚少。因此,我們利用定量蛋白質(zhì)組學(xué)技術(shù)試圖深入的研究子癇前期子代心血管功能紊亂的分子機(jī)制。iTRAQ結(jié)果顯示我們總共鑒定到1521個(gè)蛋白質(zhì),其中1496個(gè)蛋白質(zhì)具有定量信息。進(jìn)一步分析得到在臍動(dòng)脈血管中有53個(gè)具有顯著差異表達(dá)的蛋白質(zhì),其中22個(gè)蛋白質(zhì)上調(diào)表達(dá),31個(gè)蛋白質(zhì)下調(diào)表達(dá)。通過利用IPA軟件進(jìn)行深入的生物信息學(xué)分析,我們發(fā)現(xiàn)了一組與心血管密切相關(guān)的蛋白質(zhì)(SLIT3,CD99,FN1,DAPK3,CRKL,FBN1,C1 QBP,CSNK2B,AKR1B1,TMOD1,LRPA P1,FBN2,PRKCA),這些蛋白質(zhì)與心血管疾病和功能密切相關(guān),功能方面主要是與血管生成(angiogenesis)最為相關(guān),相關(guān)疾病方面則是跟高血壓(hypertension)聯(lián)系最為密切。我們對AKR1B1,FN1,FBN1這三個(gè)蛋白質(zhì)進(jìn)行了western b lot的驗(yàn)證,結(jié)果與蛋白質(zhì)組學(xué)結(jié)果一致。在對這些心血管功能相關(guān)蛋白的上游調(diào)控分析中,我們發(fā)現(xiàn)miR-9-5p可以抑制AKRIB 1,FBN1,FBN2,PRKCA這四個(gè)蛋白的表達(dá)。心血管功能相關(guān)蛋白的相互作用網(wǎng)絡(luò)顯示FN1蛋白處于軸心位置。研究結(jié)果對揭示子癇前期子代的血管功能紊亂提供了新的研究視角。第二部分：基于子癇前期動(dòng)物模型的成年子代血管的蛋白質(zhì)組學(xué)分析心血管疾病的胚胎起源是全世界備受重視的研究領(lǐng)域。流行病學(xué)調(diào)查顯示子癇前期所造成的宮內(nèi)不良環(huán)境會導(dǎo)致胎兒的血管功能出現(xiàn)不同程度的紊亂,從而加大了子代發(fā)育過程中心血管疾病發(fā)生的風(fēng)險(xiǎn),但其分子機(jī)制仍有待研究。在前期研究中,我們分析了子癇前期子代的臍動(dòng)脈差異蛋白表達(dá)譜,為了更深入的研究子癇前期子代成年后的血管功能變化,我們建立了子癇前期的動(dòng)物模型,喂養(yǎng)子代至成年(1年)后,我們收集其胸主動(dòng)脈進(jìn)行定量蛋白質(zhì)組學(xué)的研究。iTRAQ結(jié)果顯示我們總共鑒定到1825個(gè)蛋白質(zhì),進(jìn)一步分析可知有106個(gè)具有顯著差異表達(dá)的蛋白質(zhì),其中75個(gè)蛋白質(zhì)上調(diào)表達(dá),31個(gè)蛋白質(zhì)下調(diào)表達(dá)。通過IPA軟件的疾病與功能分析,我們發(fā)現(xiàn)差異表達(dá)蛋白中有20個(gè)心血管疾病和功能都有明顯相關(guān)性。其中血管生成(angiogenesis)是心血管相關(guān)功能中最為密切的,相關(guān)疾病方面則是跟動(dòng)脈阻塞(occlusion of artery)和動(dòng)脈粥樣硬化(atherosclerosis)最為相關(guān)。在對這些心血管功能相關(guān)蛋白的上游調(diào)控分析中,我們發(fā)現(xiàn)miR-423-5p起到了抑制表達(dá)的作用,而TP53則是激活表達(dá)的作用。本原創(chuàng)性課題是生殖醫(yī)學(xué)和代謝性疾病發(fā)生研究結(jié)合的嶄新課題,研究成果將對人類胚胎源性心血管病發(fā)病機(jī)制的闡明作出貢獻(xiàn)。
[Abstract]:The first part was based on the proteomic analysis of the umbilical artery of pre-eclampsia based on iTRAQ-2D-LC-MS/MS, which showed that preeclampsia had a significant impact on maternal and fetal health. Adverse conditions in the uterus may lead to cardiovascular disease in adults, but little is known about its molecular mechanism. Therefore, we use quantitative proteomics to study the molecular mechanism of cardiovascular dysfunction in pre-eclampsia. ITRAQ results show that we have identified a total of 1521 proteins, 1496 of which have quantitative information. Further analysis revealed that there were 53 differentially expressed proteins in the umbilical artery, 22 of which were up-regulated and 31 were down-regulated. By using IPA software for further bioinformatics analysis, we found a group of proteins closely related to cardiovascular diseases, such as SLIT3, CD9FN1, DAPK3, CRKL, FBN1C 1, QBP1, CSNK2B1, AKR1B1, TMOD1, LRPA, FBN2, PRKCAA, these proteins are closely related to cardiovascular disease and function. The functional aspect is mainly related to angiogenesis, while the related disease is most closely related to hypertension hypertension. Three proteins, AKR1B1FN1FBN1, were confirmed by western b lot, and the results were in agreement with the results of proteomics. In the upstream regulatory analysis of these cardiovascular function-related proteins, we found that miR-9-5p could inhibit the expression of four proteins, AKRIB 1, FBN 1, FBN 2 and PRKCA. The interaction network of cardiovascular function-related proteins shows that the FN1 protein is in an axial position. The results provide a new perspective for revealing vascular dysfunction in preeclampsia. Part two: proteomics of adult offspring based on animal model of preeclampsia is a very important research field in the world. Epidemiological investigation shows that the adverse environment caused by preeclampsia may lead to different degrees of disorder of fetal vascular function, which increases the risk of cardiovascular disease during the development of offspring, but its molecular mechanism remains to be studied. In the previous study, we analyzed the differential protein expression profiles of umbilical artery in pre-eclampsia. In order to further study the changes of vascular function in pre-eclampsia, we established an animal model of pre-eclampsia. After feeding the offspring to adulthood (1 year), we collected the thoracic aorta for quantitative proteomics. The results of iTRAQ showed that we identified a total of 1825 proteins. Further analysis revealed that 106 proteins were significantly differentially expressed. Among them, 75 proteins were up-regulated and 31 proteins were down-regulated. By analyzing the disease and function of IPA software, we found that 20 of the differentially expressed proteins were related to cardiovascular disease and function. Angiogenesis-angiogenesis is the most closely related cardiovascular function, and related diseases are most closely related to occlusion of artery occlusion and atherosclerosis. In the upstream regulatory analysis of these cardiovascular function-related proteins, we found that miR-423-5p inhibits expression and TP53 activates expression. This original subject is a new research topic of reproductive medicine and metabolic diseases. The research results will contribute to the elucidation of the pathogenesis of human embryonic cardiovascular disease.
【學(xué)位授予單位】：浙江大學(xué)
【學(xué)位級別】：博士
【學(xué)位授予年份】：2016
【分類號】：R714.244

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 桑翠琴;張震宇;王淑珍;劉浩;郭淑麗;;重度子癇前期/子癇合并急性呼吸窘迫綜合征的診斷與處理[J];中國醫(yī)刊;2006年10期

2 趙桂花;;重度子癇前期與子癇82例臨床分析[J];臨床醫(yī)藥實(shí)踐雜志;2007年01期

3 楊小頎;嚴(yán)妮子;;子癇前期與血鈣水平相關(guān)性研究[J];實(shí)用臨床醫(yī)學(xué);2007年04期

4 楊小頎;嚴(yán)妮子;;子癇前期與血鈣水平相關(guān)性研究[J];四川生理科學(xué)雜志;2007年01期

5 陳茜;王澤華;;子癇前期的預(yù)測及預(yù)防[J];華中醫(yī)學(xué)雜志;2007年03期

6 朱倩;;對38例子癇前期(重)、子癇治愈的體會[J];實(shí)用預(yù)防醫(yī)學(xué);2007年05期

7 梁英;周麗麗;甘娟;姜碧洋;楊承東;;重度子癇前期在產(chǎn)科ICU監(jiān)護(hù)救治的臨床分析[J];中國婦幼保健;2008年19期

8 詹衛(wèi)星;鄭九生;;血管緊張素轉(zhuǎn)換酶基因多態(tài)性與重度子癇前期及腎功能損害的關(guān)系[J];江西醫(yī)藥;2008年08期

9 蔡穎;尹國武;姜鋒;李怡;閔保華;王s，

本文編號：1875096