本文選題：蛋白質(zhì)組學(xué) + 多發(fā)性肌炎��； 參考：《福建醫(yī)科大學(xué)》2015年碩士論文

【摘要】：【目的】尋找可預(yù)測多發(fā)性肌炎(Polymyositis,PM)及合并肺間質(zhì)病變(Interstitial lung disease,ILD)疾病發(fā)生和發(fā)展的血清蛋白質(zhì)�！痉椒ā�1.病例收集:收集確診為PM患者血清3例,男性1例,女性2例,年齡為46.35±16.24y。PM+ILD患者血清3例,男性1例,女性2例,年齡為47.78±19.64y。同期健康體檢者血清3例,男性1例,女性2例,年齡為45.66±17.62y。三組人員的年齡,性別皆相匹配。2.三組血清在相同條件下進(jìn)行三次雙向凝膠電泳分離蛋白質(zhì)點(diǎn),考馬斯亮蘭染色后,各組凝膠用Image Master 2D Platinum7.0 software軟件分析凝膠圖像,三組所有的凝膠均取得清晰的2-DE表達(dá)圖譜。設(shè)置平滑(Smooth=10)、顯著值(Saliency=2)、最小區(qū)域(Min Area=5)等參數(shù)進(jìn)行斑點(diǎn)檢測,設(shè)置landmark蛋白點(diǎn)后進(jìn)行匹配。數(shù)據(jù)分析后選取光密度差異(ratio)2且差別具統(tǒng)計(jì)學(xué)意義(anova≤0.05)的蛋白質(zhì)點(diǎn)為差異表達(dá)的蛋白質(zhì)點(diǎn)。挖取差異表達(dá)蛋白質(zhì)點(diǎn),酶解后進(jìn)行基質(zhì)輔助激光解析電離飛行時(shí)間質(zhì)譜(MALDI-TOF-MS)分析,并與數(shù)據(jù)庫進(jìn)行匹配,鑒定蛋白質(zhì)。3.第二部分實(shí)驗(yàn):擴(kuò)大樣本量,入選PM組13例,年齡為47.26±15.40 y;PM+ILD組6例,年齡為47.33±19.14 y;SLE組年齡為46.8±17.05 y;健康對(duì)照組20例,年齡為44.76±13.64y,收集血清。應(yīng)用酶聯(lián)免疫吸附法(enzyme linked immunosorbent assay,ELISA)檢測三組血清中的血清淀粉樣蛋白A、凝集素、轉(zhuǎn)鐵蛋白及β2-糖蛋白1。【結(jié)果】1.第一部分:(1)PM組找到的蛋白質(zhì)點(diǎn)為252±11個(gè),PM合并ILD組找到的蛋白質(zhì)點(diǎn)235±8個(gè),健康對(duì)照組找到的蛋白質(zhì)點(diǎn)為239±7個(gè)。(2)與健康對(duì)照組比較,PM組有12個(gè)差異蛋白表達(dá)上調(diào),1個(gè)差異蛋白表達(dá)下調(diào);表達(dá)上調(diào)的差異蛋白成功鑒定出5種差異蛋白即血清淀粉樣蛋白A、載脂蛋白A1、血清白蛋白、凝集素、β2-糖蛋白-1;表達(dá)下調(diào)的差異蛋白鑒定為轉(zhuǎn)鐵蛋白;PM合并ILD組有13個(gè)差異蛋白同時(shí)表達(dá)下調(diào),成功鑒定出4種蛋白質(zhì)即轉(zhuǎn)鐵蛋白、鋅-a2-糖蛋白、a2-H2-糖蛋白、a1-β-糖蛋白。(3)與PM組比較,PM合并ILD組的7個(gè)差異蛋白同時(shí)表達(dá)下調(diào)。成功鑒定出2種蛋白質(zhì)即β2-糖蛋白1和轉(zhuǎn)鐵蛋白。2.第二部分:(1)血清淀粉樣蛋白A在PM、PM+ILD、SLE、HD組的血清濃度分別為609.0 ng/L、798.5 ng/L、634.0 ng/L、617.0 ng/L,在各組間比較無顯著性差異(P0.05)。(2)凝集素在PM、PM+ILD、SLE、HD組的血清濃度分別為13.0ng/L、14.0 ng/L、11.5 ng/L、8.0 ng/L;與健康對(duì)照組比較,PM、PM+ILD、SLE表達(dá)上調(diào)(差異顯著,P0.05),其余組間比較無顯著性差異(P0.05)。(3)轉(zhuǎn)鐵蛋白在PM、PM+ILD、SLE、HD組的血清濃度分別為4.06 ng/L、4.67 ng/L、3.34 ng/L、3.45 ng/L,在各組間比較無顯著性差異(P0.05)。(4)β2-糖蛋白1在PM、PM+ILD、SLE、HD組的血清濃度分別為5.0 ng/L、5.04 ng/L、3.75 ng/L、2.29 ng/L。在各組間比較無顯著性差異(P0.05)。【結(jié)論】凝集素可作為PM疾病血清生物學(xué)標(biāo)志物的候選分子。
[Abstract]:[objective] to find out the serum proteins that can predict the occurrence and development of polymyositis (PMN) and interstitial lung disease (ILD). [methods] 1. Case collection: serum samples were collected from 3 patients with PM, male 1, female 2, age 46.35 鹵16.24y.PM ILD, male 1, female 2, age 47.78 鹵19.64y. In the same period, there were 3 healthy persons, 1 male and 2 female, the age was 45.66 鹵17.62 ys. The age and gender of the three groups matched. 2. 2. Three groups of serum were separated by three times of two-dimensional gel electrophoresis under the same conditions. After Coomassie brilliant blue staining, the gel images were analyzed by Image Master 2D Platinum 7.0 software. All the three groups obtained clear 2-DE expression patterns. The parameters such as smooth Smoothopsis 10, salience 2, minimum region Min area 5) were used for spot detection, and landmark protein points were set for matching. After data analysis, the protein spots with difference in optical density and difference between two groups were selected as differentially expressed protein spots. The differentially expressed protein spots were extracted and analyzed by matrix assisted laser desorption ionization time of flight mass spectrometry (MALDI-TOF-MSMS) after enzymatic hydrolysis. The second part of the experiment: expanded the sample size, selected PM group 13 cases, age 47.26 鹵15.40 yPM-ILD group 6 cases, age 47.33 鹵19.14 ym SLE group age 46.8 鹵17.05 yy; healthy control group 20 cases, age 44.76 鹵13.64 ys. collect serum. Serum amyloid A, agglutinin, transferrin and 尾 2-glycoprotein 1 were detected by enzyme linked immunosorbent assaysa in three groups by enzyme linked immunosorbent assay (Elisa). [results] 1. Part one: the protein spots found in PM group were 252 鹵11 protein spots and 235 鹵8 protein spots in ILD group. Compared with control group, there were 12 differentially expressed proteins and 1 down-regulated protein expression in PM group. Five differentially expressed proteins, namely serum amyloid protein A, apolipoprotein A1 and serum albumin, were successfully identified. The down-regulated differential protein was identified as 13 differentially expressed proteins in transferrin PM combined with ILD group, and four proteins, transferrin, were successfully identified. Compared with PM group, the expression of 7 differentially expressed proteins in PM combined with ILD group was down-regulated. Two proteins, 尾 2 glycoprotein 1 and transferrin 2, were successfully identified. Part 2: 1) the serum concentrations of amyloid A in the PMPM ILDE-SLEHD group were 609.0 ng / L, 798.5 ng / L, 634.0 ng / L, 617.0 ng / L, respectively, and there was no significant difference among the three groups (P0.05. 2) the serum concentration of amyloid A in the PMPM ILD SLEHD group was 13.0ng / L = 14.0 ngL / L = 8.0ngL / L, respectively, as compared with that in the healthy control group (P < 0.05 / L = 8.0ng / L = 8.0ngL / L = 8.0ngL / L = 8.0ng / L = 8.0ng / L = 8.0ng / L = 8.0ng / L). The serum concentration of transferrin in PMPM ILDN SLEHD group was 4.06 ng / L 4.67 ng / L 3.34 ng / L 3.45 ng / L respectively. There was no significant difference between the two groups in 尾 _ 2-glycoprotein 1 尾 _ 2-glycoprotein _ 1 concentration in the PMPM ILDD-SLEHD group. There was no significant difference between the two groups in serum 尾 _ 2-glycoprotein _ (1) levels in the PMPM ILD SLEHD group (P _ (0.05) P _ (0.05) P _ (0.05) P _ (0.05) P _ (3.45) ng / L). The serum levels of transferrin 1 in the PMPM ILD SLEHD group were 4.06 ng / L ~ (4.67) ng / L ~ (3.34) ng / L ~ (3.45) ng / L, respectively. The concentration was 5.0 ng / L 5.04 ng / L 3.75 ng / L = 2.29 ng / L respectively. There was no significant difference among the groups. [conclusion] Lectin can be used as a candidate molecule for serum biomarkers of PM disease.
【學(xué)位授予單位】：福建醫(yī)科大學(xué)
【學(xué)位級(jí)別】：碩士
【學(xué)位授予年份】：2015
【分類號(hào)】：R593.2;R563

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