本文選題：COPD + 蛋白質(zhì)組學��； 參考：《復旦大學》2012年博士論文

【摘要】：第一部分慢性阻塞性肺病蛋白質(zhì)組學相關(guān)文獻回顧 目的：蛋白質(zhì)組學方法用于研究細胞、活檢組織、體液中的蛋白組分,廣泛用于深入探索人類疾病機制,尋找用于診斷、治療、預后的特異性的新型生物標志物,以及研發(fā)藥物靶位等方面。本文第一部分研究意在通過回顧以往慢性阻塞性肺病(chronic obstructive pulmonary disease, COPD)的蛋白質(zhì)組學報道,從中總結(jié)已有的候選生物標志物。 方法：利用文獻檢索,回顧以往基于蛋白質(zhì)組學方法的COPD報道。歸納總結(jié)這些報道涉及的研究對象、研究方法、候選生物標志物等內(nèi)容。同時比較部分候選生物標志物在其他慢性肺部疾病的變化。 結(jié)果：通過文獻檢索,得到符合納入標準的文獻共8篇。所涉及的研究對象,除COPD外,還包括正常非吸煙人群,健康吸煙者,α1胰蛋白酶缺陷者,支氣管哮喘,肺囊性纖維化,特發(fā)性肺間質(zhì)纖維化,支氣管擴張等。所選用的研究標本來源于肺組織活檢標本,支氣管肺泡灌洗液,痰液或血液。使用的蛋白質(zhì)組學實驗手段包括雙向電泳,反向高效液相色譜,串聯(lián)質(zhì)譜法,基質(zhì)輔助激光解離質(zhì)譜法,表面增強激光解吸電離質(zhì)譜法等。在不同標本中發(fā)現(xiàn)的大量候選生物標志物可歸屬于氧化-抗氧化系統(tǒng),蛋白酶-抗蛋白酶系統(tǒng),炎癥介質(zhì)以及肺部特異性蛋白等。 結(jié)論：單一蛋白難以作為生物標志物,而由多個蛋白組成的生物標志物組群則可更好的反映復雜疾病的狀態(tài)。蛋白質(zhì)組學在COPD研究中,特別是挖掘生物標志物方面獨具優(yōu)勢,但仍需建立標準化體系,規(guī)范組學研究,并最大限度的聯(lián)系臨床資料；利用生物信息學工具將臨床資料和蛋白質(zhì)組學結(jié)果相聯(lián)系,以期建立數(shù)字化模式監(jiān)測COPD的進程和預后。 第二部分COPD及急性加重期病例血漿炎性生物標志物組群的定性研究 目的：COPD不斷增長的死亡率與癥狀急性加重(chronic obstructive pulmonary disease in acute exacerbatioins, AECOPD)的頻率和嚴重程度有關(guān)。系統(tǒng)性炎癥在COPD/AECOPD中發(fā)揮重要作用。在第一部分研究中發(fā)現(xiàn),目前蛋白質(zhì)組學中存在的問題之一是如何將組學的結(jié)果與臨床信息相結(jié)合。因而本文第二部分研究意在通過蛋白質(zhì)組學方法檢測多種炎性介質(zhì),并利用新方法將組學結(jié)果和臨床信息結(jié)合,以期從新角度發(fā)現(xiàn)候選生物標志物組群。 方法：收集健康正常人、COPD以及AECOPD患者的血液標本,利用抗體芯片,定性檢測507個血漿炎癥因子。設計一套用于評價COPD及AECOPD病例病情的臨床評分系統(tǒng)(DESS),包括主要癥狀、體征、實驗室檢查等臨床信息,用于評估患者疾病的嚴重程度。將芯片檢測的差異蛋白與DESS評分結(jié)合進行相關(guān)性分析。 結(jié)果：20個炎癥因子在3組人群間存在顯著差異(p0.05)。其中Cerberus1, Growth Hormone R, IL-1F6, IL-17B R, IL-17D, IL-19, Lymphotoxin β,MMP-10, Thrombopoietin和TLR4與患者DESS評分相關(guān)(p0.05)。在AECOPD患者中存在系統(tǒng)性炎癥的下調(diào)反應。 結(jié)論：20個炎癥因子可能組成檢測COPD/AECOPD的候選生物標志物組群。將臨床信息量化并與組學結(jié)果結(jié)合的方法,為篩選疾病特異性和疾病階段特異性的生物標志物提供了新的研究模式。 第三部分COPD及急性加重期病例血漿趨化因子組群動態(tài)變化的研究 目的：第三部分意在利用蛋白質(zhì)組學方法定量檢測COPD/AECOPD循環(huán)中趨化因子的動態(tài)變化。并繼續(xù)使用第二部分中的評分系統(tǒng),對AECOPD患者進行動態(tài)評分,以期發(fā)現(xiàn)與AECOPD病程進展相關(guān)的候選生物標志物組群。 方法：收集健康正常人、COPD以及AECOPD患者的血液標本。AECOPD患者分別在確診時、治療中及癥狀好轉(zhuǎn)后采集血液標本。利用多重趨化因子抗體芯片,定量檢測血漿中40個趨化因子。將芯片檢測的差異蛋白與DESS評分結(jié)合進行相關(guān)性分析。 結(jié)果：在40個趨化因子中,30個在COPD和健康正常人間存在統(tǒng)計學差異(p0.05),16個在AECOPD和健康正常人間存在統(tǒng)計學差異(p0.05)。13個趨化因子,包括BTC,IL-9, IL-18Bpa, CCL22, CCL23, CCL25, CCL28, CTACK, LIGHT, MSPa, MCP-3, MCP-4和OPN,在三組間均存在差異。部分趨化因子的表達量與DESS評分間存在關(guān)聯(lián)(p0.05)。 結(jié)論：13個趨化因子可能組成監(jiān)測AECOPD發(fā)生和治療進展的候選生物標志物組群。第二部分中的研究方法,為篩選疾病特異性和疾病階段特異性的生物標志物提供了新的研究模式。
[Abstract]:Literature Review on the Proteomics of Chronic Obstructive Pulmonary Disease in Part I

Objective : Proteomics method is used to study the protein components of cells , biopsy tissues and body fluids . It is widely used to probe into the mechanism of human diseases , to find novel biomarkers for diagnosis , treatment and prognosis , as well as to research and develop drug targets . The first part of this paper is to summarize the existing candidate biomarkers by reviewing the proteomic report of chronic obstructive pulmonary disease ( COPD ) .

Methods : Literature search was used to review the previous COPD reports based on proteomic methods . The subjects , methods and candidate biomarkers of these reports were summarized . At the same time , the changes of some candidate biomarkers in other chronic lung diseases were compared .

Results : A total of 8 articles conforming to the standard were obtained by literature search . In addition to COPD , there were normal non - smokers , healthy smokers , 偽 1 trypsin deficiency , bronchial asthma , pulmonary cystic fibrosis , idiopathic pulmonary fibrosis , bronchiectasis , etc . The selected study samples were derived from lung tissue biopsy specimens , bronchoalveolar lavage fluid , sputum or blood . The methods used in the study included two - dimensional electrophoresis , reverse high performance liquid chromatography , tandem mass spectrometry , matrix assisted laser desorption mass spectrometry , surface enhanced laser desorption ionization mass spectrometry , etc . The large number of candidate biomarkers found in different specimens could be attributed to oxidation - oxidation resistant systems , protease - antiprotease systems , inflammatory mediators , and lung - specific proteins .

Conclusion : Single protein is difficult to be used as biomarker , and biomarker group composed of multiple proteins can better reflect the state of complex disease . Proteomics has unique advantages in the study of COPD , especially in mining biomarkers , but it is still necessary to establish a standardized system , standardize the study of group and maximize the clinical data ;
A bioinformatic tool is used to link clinical data and proteomic results with a view to establishing a digital pattern to monitor the progression and prognosis of COPD .

Qualitative study of plasma inflammatory biomarkers in patients with COPD and acute exacerbation

Objective : To investigate the frequency and severity of chronic obstructive pulmonary disease in COPD / AECOPD . Systemic inflammation plays an important role in COPD / AECOPD .

Methods : Blood samples from healthy individuals , COPD and AECOPD patients were collected . 507 plasma inflammatory factors were qualitatively detected by using antibody chip . A set of clinical scoring system ( DESS ) was designed to evaluate the condition of COPD and AECOPD .

Results : There was a significant difference among the 20 inflammatory factors among the 3 groups ( p < 0.05 ) . Among them , Cerberus 1 , Growth Hormone R , IL - 1F6 , IL - 17B R , IL - 17 , IL - 19 , lymphotoxin 尾 , MMP - 10 , Thrombopoietin and TLR were correlated with the DESS score of the patients ( p < 0.05 ) . There was a down - regulation reaction of systemic inflammation in patients with AECOPD .

Conclusion : 20 inflammatory factors may constitute a candidate biomarker group for COPD / AECOPD . The clinical information is quantified and combined with the results of group learning to provide a new research mode for screening biomarkers specific to disease specificity and disease stage .

Study on the Dynamic Changes of Plasma Chemokine Cluster in the Third Part of COPD and Acute Aggravated Period

Objective : The third part is to quantitatively detect the dynamic changes of chemokine in COPD / AECOPD by proteomic method , and continue to use the scoring system in the second part to dynamically score AECOPD patients with a view to finding the candidate biomarker group associated with the progression of AECOPD .

Methods : Blood samples from healthy subjects , COPD and AECOPD patients were collected . Blood samples were collected from the patients with AECOPD .

Results : There were statistically significant differences ( p . 05 ) between 30 COPD patients and healthy controls ( p . 05 ) . There were 13 chemokine ( p . 05 ) in AECOPD and normal controls . There was a difference between the three groups . There was a correlation between the expression of partial chemokine and DESS score ( p < 0 . 05 ) .

Conclusion : 13 chemokine may constitute a candidate biomarker group for monitoring the development and progression of AECOPD . In the second part , a new research mode is provided for screening biomarkers specific to disease - specific and disease - stage .

【學位授予單位】：復旦大學
【學位級別】：博士
【學位授予年份】：2012
【分類號】：R563.9

【參考文獻】

相關(guān)期刊論文 前1條

1 蔡珊,陳平,朱應群,彭紅,鄭東元,劉志軍,蔣惜念,劉友文;慢性阻塞性肺疾病氣道炎癥與肺泡巨噬細胞炎癥蛋白1α、明膠酶B活性的研究[J];中華結(jié)核和呼吸雜志;2001年07期

，

本文編號：1776793