本文選題：腎小球 + 腎髓質(zhì)　； 參考：《北京協(xié)和醫(yī)學(xué)院》2017年博士論文

【摘要】：研究背景:基于質(zhì)譜的蛋白質(zhì)組學(xué)技術(shù)已成為探索腎臟疾病發(fā)病機(jī)制及某些疑難腎臟疾病診斷的方法之一。目前利用激光顯微切割聯(lián)合質(zhì)譜(Laser microdissection and mass spectromy,LMD/MS)技術(shù)對人腎小球進(jìn)行蛋白質(zhì)表達(dá)譜的研究有限,對腎髓質(zhì)進(jìn)行蛋白質(zhì)組學(xué)表達(dá)譜的研究尚未見發(fā)表。采用LMD/MS技術(shù)不僅可分別獲取腎小球及腎髓質(zhì)組織以構(gòu)建蛋白表達(dá)譜,也可對腎病變組織區(qū)域進(jìn)行分析。近年來,利用LMD/MS技術(shù)發(fā)現(xiàn)一些新的淀粉樣變亞型,使得淀粉樣變分型診斷水平有所提高;同時(shí)對致淀粉樣變蛋白進(jìn)行氨基酸序列分析有可能發(fā)現(xiàn)變異的致淀粉樣變蛋白。研究方法:在第一部分工作中,我們收集6例癌旁腎組織,通過LMD/MS技術(shù)獲取腎小球及腎髓質(zhì)組織并構(gòu)建蛋白質(zhì)組表達(dá)譜;對兩個(gè)部位所鑒定到的蛋白進(jìn)行生物信息學(xué)分析。在第二部分工作中,我們分析了近5年45例AL型淀粉樣變性病患者臨床、實(shí)驗(yàn)室檢查及腎活檢病理結(jié)果,選取20例腎組織免疫熒光分型不明確的患者進(jìn)行免疫組化及LMD/MS分型,評價(jià)免疫組化及LMD/MS在免疫熒光分型不明確的AL淀粉樣變患者中的診斷意義。在第三部分工作中,對9例臨床、實(shí)驗(yàn)室檢查及腎組織免疫化學(xué)檢查均不能分型的疑難腎淀粉樣變患者進(jìn)行LMD/MS分析,以期達(dá)到腎淀粉樣變的完整分型診斷,并利用生物信息學(xué)方法對AA型淀粉樣變病例的SAA蛋白進(jìn)行變異氨基酸序列分析。研究結(jié)果:第一部分中,腎小球及腎髓質(zhì)分別鑒定到1373及1673種蛋白;與以往研究比較,本研究單獨(dú)鑒定到472種腎小球蛋白,并首次建立了正常人類腎髓質(zhì)蛋白質(zhì)組表達(dá)譜。生物信息學(xué)分析顯示腎小球蛋白多為細(xì)胞骨架蛋白,腎髓質(zhì)蛋白多參與細(xì)胞代謝、具有酶解功能。第二部分中,我們發(fā)現(xiàn)應(yīng)用免疫組化方法可對免疫熒光不能分型的部分AL型淀粉樣變進(jìn)行分型診斷,敏感性約為55%;對于部分免疫組化方法不能分型的AL型淀粉樣變患者,LMD/MS技術(shù)仍可進(jìn)行分型診斷;20例行LMD/MS分析的患者中,18例可明確分型診斷,敏感性為90%。第三部分中,9例疑難腎淀粉樣變分型不明確的患者,經(jīng)LMD/MS診斷出4例AL型淀粉樣變,1例Apo AIV型淀粉樣變,1例AA型淀粉樣變及1例AH型淀粉樣變;AA型淀粉樣變患者淀粉樣物質(zhì)中包含SAA1及SAA2蛋白,未發(fā)現(xiàn)兩種蛋白的氨基酸序列變異。研究結(jié)論:本研究利用LMD/MS技術(shù),對新鮮冰凍腎組織顯微切割獲取的腎小球及腎髓質(zhì)組織進(jìn)行蛋白質(zhì)組學(xué)及生物信息學(xué)分析,豐富了腎小球蛋白質(zhì)表達(dá)譜數(shù)據(jù)庫,并首次建立了正常人類腎髓質(zhì)蛋白質(zhì)組表達(dá)譜。免疫組化染色及LMD/MS分析發(fā)現(xiàn),對于免疫熒光不能分型的腎AL型淀粉樣變病例,采用免疫組化的方法可改進(jìn)分型診斷的水平。LMD/MS技術(shù)對于部分免疫化學(xué)方法不能分型的腎AL型淀粉樣變病例可提供分型診斷,該技術(shù)能鑒定到淀粉樣纖維的多種蛋白成分,也適用于少見類型的淀粉樣變性的分型診斷并確證免疫熒光或免疫組化結(jié)果,具有一定的臨床應(yīng)用前景。
[Abstract]:Research background: proteomics mass spectrometry has become one of the methods to explore the pathogenesis of kidney disease and some difficult diagnosis of kidney disease. Based on the current using laser microdissection combined with mass spectrometry (Laser microdissection and mass spectromy, LMD/MS) on human renal small ball protein expression studies were limited, learn expression study is published no renal medulla of protein group. Using LMD/MS technology can not only obtain the glomerular and renal medullary tissue to construct protein expression profile, but also on the renal lesion area is analyzed. In recent years, the use of LMD/MS technology to find some new amyloidosis subtypes, the amyloid type diagnostic level increased; at the same time the amyloidogenic protein amino acid sequence analysis is likely to find the variation of amyloidogenic proteins. Methods: in the first part, I We collected 6 cases of adjacent kidney glomerulus and renal medullary tissue obtained by LMD/MS technology and construct the proteome expression profile; bioinformatics analysis identified two sites of protein. In the second part, we analyzed the past 5 years in 45 cases of AL amyloidosis patients with clinical laboratory. Check and renal biopsy results from immunofluorescence in 20 cases of renal tissue typing is not clear with immunohistochemistry and LMD/MS classification, evaluation of immunohistochemistry and immunofluorescence typing in LMD/MS is not clear diagnosis of AL amyloidosis patients. In the third part, on 9 cases. LMD/MS analysis of patient examination and kidney tissue laboratory immunohistochemical examination were not types of difficult renal amyloid, renal amyloidosis in order to achieve the complete classification, and the use of biological information for AA amyloidosis cases methodology S Analyze the variation of amino acid sequence of AA protein. Results: in the first part, glomerular and renal medulla were identified to 1373 and 1673 proteins; and a comparison with former studies, this study identified 472 separate glomerular protein, established the normal human renal medullary proteome expression profile. Bioinformatics analysis showed glomerular protein cytoskeletal protein, renal medulla protein involved in cell metabolism, with enzymatic functions. In the second part, we found that using immunohistochemical method of immunofluorescence typing can not be part of AL amyloidosis in the differential diagnosis, the sensitivity is about 55%; the immunohistochemical method not classified patients with AL amyloidosis, LMD/MS technology can be classified diagnosis; analysis of 20 cases of LMD/MS patients, 18 cases of definite diagnosis of type, the sensitivity of 90%. in the third part, 9 cases of difficult renal amyloidosis Typing is not clear in patients after LMD/MS diagnosed 4 cases of AL amyloidosis, 1 cases of Apo AIV amyloidosis, 1 cases of AA amyloidosis and 1 cases of AH amyloidosis; AA amyloidosis include SAA1 and SAA2 protein in patients with amyloid, found no amino acid sequence two kinds of protein variation. Conclusion: in this study, using LMD/MS technology, proteomics and bioinformatics analysis of glomerular and renal medullary tissues of fresh frozen kidney tissue microdissection to obtain, enrich the glomerular protein expression database, established the normal human renal medullary proteome expression profile. Immunohistochemistry LMD/MS staining and immunofluorescence analysis showed that, for not classified renal AL amyloidosis cases by immunohistochemical method can improve renal AL type starch levels.LMD/MS typing typing for not part of immunochemical method Variant cases can provide typing diagnosis. This technology can identify various protein components of amyloid fibrils, and also can be applied to the typing diagnosis of rare types of amyloidosis and confirm the results of immunofluorescence or immunohistochemistry. It has certain clinical application prospects.

【學(xué)位授予單位】：北京協(xié)和醫(yī)學(xué)院
【學(xué)位級別】：博士
【學(xué)位授予年份】：2017
【分類號】：R692

【相似文獻(xiàn)】

相關(guān)期刊論文 前10條

1 吳小煉;以腹瀉為主要表現(xiàn)淀粉樣變長期誤診1例[J];中國綜合臨床;2000年09期

2 劉紅春,張順財(cái);淀粉樣變[J];肝臟;2000年03期

3 史玉玲;何桂蘭;;異維A酸治療原發(fā)性皮膚淀粉樣變臨床觀察[J];醫(yī)學(xué)研究通訊;2001年03期

4 劉小軍,潘康儉;原發(fā)性皮膚淀粉樣變109例臨床和病理分析[J];皮膚病與性病;2002年04期

5 鄧安國,蘇華;繼發(fā)性腎淀粉樣變的發(fā)病機(jī)制與治療[J];臨床內(nèi)科雜志;2002年06期

6 李雁,彭德榮;異維A酸治療32例原發(fā)性皮膚淀粉樣變臨床觀察[J];中國廠礦醫(yī)學(xué);2005年03期

7 Parsi K. ,Kossard S. ,張憲旗;熱敏苔癬狀淀粉樣變[J];世界核心醫(yī)學(xué)期刊文摘(皮膚病學(xué)分冊);2005年05期

8 李別非,江亞文;系統(tǒng)性淀粉樣變1例[J];中國老年學(xué)雜志;2005年11期

9 李祖海;余英豪;;淀粉樣變誤診為原發(fā)性腎病綜合征2例報(bào)告[J];安徽醫(yī)學(xué);2006年05期

10 劉瑩;張輝;倪然;張?bào)?;系統(tǒng)性淀粉樣變26例臨床分析[J];醫(yī)藥論壇雜志;2010年14期

相關(guān)會議論文 前10條

1 ;系統(tǒng)性淀粉樣變1例[A];2005年浙江皮膚性病學(xué)學(xué)術(shù)年會論文匯編[C];2005年

2 呂中法;;皮膚淀粉樣變?[A];2009年浙江省皮膚病學(xué)術(shù)會議論文匯編[C];2009年

3 房麗華;蔣明;;原發(fā)性干燥綜合征合并淀粉樣變?nèi)齕A];中華醫(yī)學(xué)會全國風(fēng)濕病學(xué)年會論文匯編[C];2003年

4 徐凌;蔡柏薔;鐘旭;朱元玨;;淀粉樣變的呼吸系統(tǒng)表現(xiàn)59例分析[A];中華醫(yī)學(xué)會第七次全國呼吸病學(xué)術(shù)會議暨學(xué)習(xí)班論文匯編[C];2006年

5 彭建中;;局限性結(jié)節(jié)性原發(fā)性皮膚淀粉樣變1例[A];2008全國中西醫(yī)結(jié)合皮膚性病學(xué)術(shù)會議論文匯編[C];2008年

6 蔡綏R，

本文編號：1768644