本文選題：神經(jīng)精神狼瘡　切入點：MALDI-TOF-MS　出處：《北京協(xié)和醫(yī)學(xué)院》2009年博士論文　論文類型：學(xué)位論文

【摘要】：第一部分：神經(jīng)精神狼瘡腦脊液蛋白質(zhì)指紋圖譜及分析 目的篩選神經(jīng)精神狼瘡(NPSLE)腦脊液生物標(biāo)志物,并建立相應(yīng)NPSLE疾病的樹形分類模型。材料和方法用MALDI-TOF-MS聯(lián)合WCX磁珠生成腦脊液蛋白質(zhì)譜,比較NPSLE治療前組(27例)與脊柱側(cè)凸組(17例)、無神經(jīng)精神癥狀的系統(tǒng)性紅斑狼瘡組(non-NPSLE,10例)之間的差異蛋白峰；建立NPSLE的樹形分類模型,并用其他中樞神經(jīng)系統(tǒng)受累疾病(腰椎間盤突出12例,其他結(jié)締組織病相關(guān)腦病4例)進行盲法檢驗。比較17例NPSLE患者治療前后腦脊液蛋白質(zhì)譜的差異蛋白峰。結(jié)果NPSLE治療前組與非精神神經(jīng)狼瘡組(脊柱側(cè)凸和non-NPSLE)腦脊液蛋白質(zhì)譜共有25個差異峰,用其中質(zhì)荷比為8595、7170、7661、7740和5806差異峰建立NPSLE樹形分類模型,其對源數(shù)據(jù)的敏感性92.6%,特異性92.6%；其他中樞神經(jīng)系統(tǒng)受累疾病(16例)盲法檢驗顯示該模型分類NPSLE的特異性為81.3%。17例NPSLE患者治療前后腦脊液質(zhì)譜有4個差異峰,其中質(zhì)荷比為4963和8595蛋白峰顯著降低,質(zhì)荷比為6637和6896蛋白峰顯著升高。結(jié)論MALDI-TOF-MS聯(lián)合WCX磁珠可用于腦脊液蛋白質(zhì)組研究,發(fā)現(xiàn)了多個潛在腦脊液生物標(biāo)志物可能用于NPSLE檢測、診斷和病情評估,建立了敏感性及特異性均好的NPSLE樹形分類模型。 第二部分：神經(jīng)精神狼瘡血清蛋白質(zhì)指紋圖譜及分析 目的篩選神經(jīng)精神狼瘡(NPSLE)血清生物標(biāo)志物。材料和方法用MALDI-TOF-MS聯(lián)合WCX磁珠生成血清蛋白質(zhì)譜,比較NPSLE治療前組(24例)與健康對照組(21例)、無神經(jīng)精神癥狀的狼瘡組(non-NPSLE,8例)之間的差異蛋白峰；比較13例NPSLE治療前后血清蛋白質(zhì)譜的差異蛋白峰；比較同一NPSLE患者同期血清與腦脊液質(zhì)譜相同質(zhì)荷比差異蛋白峰的相互關(guān)系。結(jié)果NPSLE治療前組與健康對照組血清蛋白質(zhì)譜有35個差異峰,與non-NPSLE組有13個差異峰；non-NPSLE組與健康對照組有15個差異峰,其中質(zhì)荷比為6438、8122、8690、9280、28081蛋白峰在上述多組之間均存在統(tǒng)計學(xué)及質(zhì)譜圖差異。13例NPSLE患者治療前后血清質(zhì)譜中質(zhì)荷比為4959、6189蛋白峰治療后升高顯著。同一NPSLE患者同期血清與腦脊液質(zhì)譜相同質(zhì)荷比差異蛋白峰共4對：4959(血清)4963(腦脊液),NPSLE治療后4959顯著升高,而4963顯著降低；5905(血清)5866(腦脊液),NPSLE治療前組均高于正常對照組；7754(血清)7740(腦脊液)和8581(血清)8595(腦脊液),均為NPSLE治療前組血清水平降低,而腦脊液水平升高。結(jié)論發(fā)現(xiàn)多個潛在血清生物標(biāo)志物可能用于神經(jīng)精神狼瘡檢測、診斷和病情評估,其中同—NPSLE患者同期血清與腦脊液質(zhì)譜相同質(zhì)荷比的4對差異蛋白峰可能更有研究和臨床應(yīng)用價值。 第三部分：神經(jīng)精神狼瘡腦脊液蛋白質(zhì)指紋圖譜差異蛋白峰鑒定 研究背景及目的血清淀粉樣蛋白A(SAA)是一種急性期蛋白,參與激活疾病的多種前炎癥反應(yīng)。多項研究鑒定血清SELDI-TOF-MS質(zhì)譜的質(zhì)荷比為11.7KD蛋白峰對應(yīng)SAA蛋白。本試驗擬鑒定腦脊液質(zhì)譜質(zhì)荷比11.7KD的差異蛋白峰為SAA蛋白。材料和方法SAA抗體進行腦脊液免疫沉淀后,分別運用MALDI-TOF-MS聯(lián)合WCX磁珠法和免疫印跡法檢測免疫沉淀反應(yīng)后上清及沉淀蛋白。結(jié)果SAA抗體免疫沉淀后上清蛋白質(zhì)譜中質(zhì)荷比11.7KD的蛋白峰強度顯著下降；SAA抗體免疫沉淀后瓊脂糖珠捕捉的蛋白在免疫印跡中出現(xiàn)分子量約為12KD的目的條帶。結(jié)論腦脊液蛋白質(zhì)譜的質(zhì)荷比11.7KD蛋白峰對應(yīng)SAA蛋白,神經(jīng)精神狼瘡(NPSLE)患者腦脊液SAA水平升高,提示SAA可能成為NPSLE的生物標(biāo)志物。
[Abstract]:Part one: protein fingerprint and analysis of cerebrospinal fluid in neuromental lupus erythematosus
Objective to screen the neuropsychiatric systemic lupus erythematosus (NPSLE) in cerebrospinal fluid biomarkers, tree classification model and established the corresponding NPSLE disease. Materials and methods of using MALDI-TOF-MS combined with WCX beads cerebrospinal fluid protein mass spectrometry generates NPSLE, before the treatment group (27 cases) and scoliosis group (17 cases), systemic lupus erythematosus group without neuropsychiatric symptoms (10 non-NPSLE cases) between different protein peaks; tree classification model based on NPSLE, and other central nervous system involvement disease (12 cases of lumbar disc herniation and other connective tissue disease related encephalopathy in 4 cases) were blind test. 17 cases of NPSLE patients before and after treatment in different protein peaks. Cerebrospinal fluid protein mass spectrometry results before NPSLE treatment group and non neuropsychiatric lupus group (scoliosis and non-NPSLE) cerebrospinal fluid protein consists of 25 distinct peaks, with the establishment of the NPSLE / Z 8595717076617740 and 5806 differential peaks The tree classification model, the sensitivity of the 92.6% sources of data, the specificity was 92.6%; the other central nervous system involvement disease (16 cases) blind test show the specificity of the NPSLE classification model for 81.3%.17 NPSLE patients before and after treatment of cerebrospinal fluid mass spectrometry 4 differential peaks, the mass ratio of 4963 and 8595 protein peaks were significantly reduced M / Z 6637, and 6896 protein peaks were significantly increased. Conclusion MALDI-TOF-MS combined with WCX can be used for beads of cerebrospinal fluid proteomic research, found a number of potential CSF biomarkers may be used for NPSLE detection, diagnosis and evaluation, established the NPSLE tree model sensitivity and specificity are good.
The second part: the fingerprint and analysis of serum protein of neuromental lupus erythematosus
Objective to screen the neuropsychiatric systemic lupus erythematosus (NPSLE) serum biomarkers. Materials and methods of using MALDI-TOF-MS combined with WCX beads generated serum protein mass spectrometry, comparison of NPSLE before the treatment group (24 cases) and control group (21 cases), lupus group without neuropsychiatric symptoms (8 cases non-NPSLE) between different protein peaks; comparison 13 cases of NPSLE between before and after treatment of serum protein protein interaction peak; compared with the same period of NPSLE patients serum and cerebrospinal fluid mass the same mass of protein peaks. The former group and the healthy control group of serum protein spectrum has 35 distinct peaks results in the treatment of NPSLE, there are 13 differences between the peak and the non-NPSLE group; non-NPSLE group and health the control group had 15 distinct peaks, including 643881228690928028081 / Z protein peaks in the group are in NPSLE patients before and after treatment of.13 and statistical difference spectra of serum in mass spectrometry 49596189 protein peaks after treatment increased significantly. The same year Xue Qing NPSLE patients with cerebrospinal fluid mass mass the same protein peaks were 4 to 4963: 4959 (Xue Qing), NPSLE (CSF) after treatment 4959 significantly increased, and 4963 decreased significantly; 5905 (Xue Qing 5866), NPSLE (CSF) before treatment group were higher than that of the normal control group; 7754 (Xue Qing) 7740 (cerebrospinal fluid) and 8581 (Xue Qing, 8595) (cerebrospinal fluid) were NPSLE group before treatment reduced the level of Xue Qing, and cerebrospinal fluid levels. Conclusion found a number of potential biomarkers for Xue Qing may God by the spirit of lupus detection, diagnosis and evaluation, of which 4 of the difference NPSLE with the same period of Xue Qing and the same mass mass cerebrospinal fluid protein peaks may be more research and clinical application value.
The third part: identification of differential protein peaks of protein fingerprints in cerebrospinal fluid of neuromental lupus erythematosus
Background and objective: serum amyloid A (SAA) is an acute phase protein, is involved in many inflammatory diseases. Many studies identified the activation of serum SELDI-TOF-MS MS / Z 11.7KD protein peaks corresponding to SAA protein. The aim of this study is to identify the different mass mass 11.7KD cerebrospinal fluid protein peak is SAA protein. SAA antibody material and the methods of cerebrospinal fluid after immunoprecipitation, respectively using MALDI-TOF-MS WCX combined detection of immune magnetic beads and Western blotting precipitation supernatant and protein precipitation after the reaction. Results the protein peaks of the supernatant protein mass spectrometry SAA antibodies after immunoprecipitation in mass of 11.7KD decreased significantly; SAA antibodies after immunoprecipitation agarose beads to capture protein in immunoblots in the molecular weight of about 12KD to Z band. 11.7KD protein peaks of SAA protein in cerebrospinal fluid protein. Conclusion neuropsychiatric systemic lupus erythematosus (NPSL E) the level of SAA in the cerebrospinal fluid (CSF) increases, suggesting that SAA may be a biomarker for NPSLE.

【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】：2009
【分類號】：R593.241;R3411

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