發(fā)布時間：2018-04-16 22:09

  本文選題：iTRAQ技術(shù) + 造影劑急性腎損傷��； 參考：《寧波大學(xué)》2017年碩士論文

【摘要】：目的:應(yīng)用同位素標(biāo)記相對和絕對定量(Isobaric Tags for Relative and Absolute Quantitation,iTRAQ)技術(shù)聯(lián)合液相色譜-串聯(lián)質(zhì)譜(Liquid Chromatography tandem Mass Spectrometry,LC-MS/MS)技術(shù)篩選和鑒定經(jīng)皮冠狀動脈介入術(shù)(percutaneous coronary intervention,PCI)后發(fā)生造影劑急性腎損傷(contrast-induced acute kidney injury,CI-AKI)患者尿液差異蛋白,尋找CI-AKI早期診斷生物標(biāo)記物。方法:收集2015年7月1日至2015年12月31在寧波市第二醫(yī)院行PCI術(shù)前及術(shù)后6h、24h、48h患者尿液,根據(jù)2012KDIGO急性腎損傷(acute kidney injury,AKI)診斷標(biāo)準(zhǔn),將患者分為:(1)CI-AKI組(PCI術(shù)后發(fā)生AKI);(2)陰性組。應(yīng)用iTRAQ技術(shù)比較CIAKI組術(shù)后6h、24h、48h與術(shù)前以及CI-AKI組術(shù)后24h與陰性組術(shù)后24h的差異蛋白。將篩選出的差異蛋白按照差異倍數(shù)排序,選出每組差異倍數(shù)前10位中與CI-AKI相關(guān)的差異蛋白進(jìn)行GO(Gene ontology)富集分析,了解新型生物標(biāo)志物在CI-AKI可能的發(fā)生機(jī)制,并進(jìn)行多組數(shù)據(jù)進(jìn)行交集分析,尋找CI-AKI潛在的生物標(biāo)志物。結(jié)果:48例行PCI術(shù)患者中有6例發(fā)生CI-AKI,4例男性,2例女性。(1)CI-AKI組術(shù)后6h:較術(shù)前差異蛋白151個,上調(diào)74個,下調(diào)77個;差異倍數(shù)前10位與CI-AKI相關(guān)的差異蛋白為免疫球蛋白G2重鏈(IGHG2)、人甘露糖結(jié)合凝集素相關(guān)絲氨酸蛋白酶2(MASP2)、血清載體蛋白A1(APOA1)、人過氧化物還原酶5(PRDX5)。(2)CI-AKI組術(shù)后24h:1)較術(shù)前差異蛋白167個,上調(diào)106個,下調(diào)61個;差異倍數(shù)前10位與CI-AKI相關(guān)的差異蛋白為癌細(xì)胞分裂周期蛋白(CDC42)、人類β防御素(HBD)、人過氧化物酶2(PRDX2)。2)較陰性組術(shù)后24h差異蛋白191個,上調(diào)77個,下調(diào)114個;差異倍數(shù)前10位與CI-AKI相關(guān)的差異蛋白為:PRDX2、水通道蛋白1(AQP1)、血清結(jié)合珠蛋白(HP)、人乳鐵蛋白(LTF)、HBD。(3)CI-AKI組術(shù)后48h組:較術(shù)前差異蛋白160個,上調(diào)105個,下調(diào)55個;差異倍數(shù)前10位與CI-AKI相關(guān)的差異蛋白為AQP1、人肽酶抑制因子(PI16)。(4)通過交集分析,結(jié)果用韋恩圖表示。結(jié)果顯示CI-AKI組各時段與術(shù)前比較的差異蛋白以及CI-AKI組術(shù)后24h與陰性組術(shù)后24h共有的差異蛋白有14個,其中與CI-AKI相關(guān)的差異蛋白為AQP1、HBD-1、MASP2。以上都是定性(5)差異蛋白GO富集分析結(jié)果:1)分子功能:主要屬于結(jié)合各種類型蛋白(包括受體)和酶的調(diào)控等功能;2)生物學(xué)過程:主要參與代謝、細(xì)胞活動和參與應(yīng)激反應(yīng)等過程;3)細(xì)胞定位:主要屬于細(xì)胞外區(qū)域、囊泡、細(xì)胞器膜等區(qū)域。結(jié)論:(1)iTRAQ技術(shù)能夠進(jìn)行絕對和相對定量,檢測結(jié)果精確并具有良好的可重復(fù)性,彌補(bǔ)了傳統(tǒng)技術(shù)的不足。能有效篩選出CI-AKI差異蛋白。(2)AQP1、HBD-1、MASP2在CI-AKI患者尿液中含量發(fā)生顯著上調(diào),三者與CI-AKI發(fā)病機(jī)制有關(guān),是潛在的新型生物標(biāo)志物。
[Abstract]:Objective: to screen and identify acute renal contrast agent after percutaneous coronary intervention with Isobaric Tags for Relative and Absolute quantity iTRAQ and liquid Chromatography tandem Mass spectrometryLC-MS / MS technique.Urine differential proteins in patients with contrast-induced acute kidney injury-CI-AKI,To search for biomarkers for early diagnosis of CI-AKI.Methods: urine samples were collected from the second Hospital of Ningbo City from July 1, 2015 to December 31, 2015. According to the diagnostic criteria of acute kidney injury-AKI before and after PCI, the patients were divided into two groups.ITRAQ technique was used to compare the differential proteins between the CIAKI group and the pre-operation group and the CI-AKI group and the negative group at 6 hours and 24 hours after the operation.The differential proteins in the first 10 positions of each group of differential multiples were selected for GO(Gene ontology enrichment to understand the possible mechanism of the new biomarkers in CI-AKI.A number of groups of data were analyzed to search for potential biomarkers of CI-AKI.Results six of the 48 patients underwent PCI had CI-AKI in 4 males and 2 females in the CI-AKI group: compared with 151 patients with preoperative differential protein, 74 patients were up-regulated and 77 patients were down-regulated.The first 10 differential proteins associated with CI-AKI were immunoglobulin G2 heavy chain IGHG2, human mannose binding agglutinin associated serine protease 2MASP21, serum carrier protein A1 APOA1, human peroxidase reductase 5(PRDX5).(2)CI-AKI group 24 h after operation.The first 10 differential proteins related to CI-AKI were cancer cell cycle protein CDC42, human 尾 -defensin HBDG and human peroxidase 2PRDX2 路2) compared with negative group at 24 hours after operation, the differential proteins were up regulated by 77 and down regulated by 114.The first 10 differential proteins related to CI-AKI were: 1 PRDX2, 1 aquaporin 1 aquaporin, HP1, HP5, HBD. 3CI-AKI: 160 proteins, 105 up-regulated and 55 down-regulated compared with those before operation.The first 10 differentially expressed proteins associated with CI-AKI were AQP1 and human peptidase inhibitor PI16. 4) the results were expressed by Wayne map.The results showed that there were 14 differentially expressed proteins in CI-AKI group and 24 hours after operation in CI-AKI group and negative group respectively. The difference protein associated with CI-AKI was AQP1HBD-1mASP2.These are qualitative differential protein (5) differential protein enrichment analysis results: 1) molecular function: mainly belong to various types of protein (including receptor) and enzyme regulation and other functions of the biological process: mainly involved in metabolism,Cellular activity and involvement in stress response were mainly located in extracellular region, vesicle, organelle and other regions.Conclusion the technique can be used in absolute and relative quantitative analysis, and the detection results are accurate and reproducible, which make up for the deficiency of traditional technique.The CI-AKI differential protein, AQP1, HBD-1MASP2 was significantly up-regulated in the urine of CI-AKI patients, which was related to the pathogenesis of CI-AKI and was a potential new biomarker.
【學(xué)位授予單位】：寧波大學(xué)
【學(xué)位級別】：碩士
【學(xué)位授予年份】：2017
【分類號】：R541.4;R692

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