多发性肌炎合并肺间质病变的血清蛋白质组学分析

发布时间：2018-06-06 22:49

本文选题：蛋白质组学 + 多发性肌炎　；参考：《福建医科大学》2015年硕士论文

【摘要】：【目的】寻找可预测多发性肌炎(Polymyositis,PM)及合并肺间质病变(Interstitial lung disease,ILD)疾病发生和发展的血清蛋白质。【方法】1.病例收集:收集确诊为PM患者血清3例,男性1例,女性2例,年龄为46.35±16.24y。PM+ILD患者血清3例,男性1例,女性2例,年龄为47.78±19.64y。同期健康体检者血清3例,男性1例,女性2例,年龄为45.66±17.62y。三组人员的年龄,性别皆相匹配。2.三组血清在相同条件下进行三次双向凝胶电泳分离蛋白质点,考马斯亮兰染色后,各组凝胶用Image Master 2D Platinum7.0 software软件分析凝胶图像,三组所有的凝胶均取得清晰的2-DE表达图谱。设置平滑(Smooth=10)、显著值(Saliency=2)、最小区域(Min Area=5)等参数进行斑点检测,设置landmark蛋白点后进行匹配。数据分析后选取光密度差异(ratio)2且差别具统计学意义(anova≤0.05)的蛋白质点为差异表达的蛋白质点。挖取差异表达蛋白质点,酶解后进行基质辅助激光解析电离飞行时间质谱(MALDI-TOF-MS)分析,并与数据库进行匹配,鉴定蛋白质。3.第二部分实验:扩大样本量,入选PM组13例,年龄为47.26±15.40 y;PM+ILD组6例,年龄为47.33±19.14 y;SLE组年龄为46.8±17.05 y;健康对照组20例,年龄为44.76±13.64y,收集血清。应用酶联免疫吸附法(enzyme linked immunosorbent assay,ELISA)检测三组血清中的血清淀粉样蛋白A、凝集素、转铁蛋白及β2-糖蛋白1。【结果】1.第一部分:(1)PM组找到的蛋白质点为252±11个,PM合并ILD组找到的蛋白质点235±8个,健康对照组找到的蛋白质点为239±7个。(2)与健康对照组比较,PM组有12个差异蛋白表达上调,1个差异蛋白表达下调;表达上调的差异蛋白成功鉴定出5种差异蛋白即血清淀粉样蛋白A、载脂蛋白A1、血清白蛋白、凝集素、β2-糖蛋白-1;表达下调的差异蛋白鉴定为转铁蛋白;PM合并ILD组有13个差异蛋白同时表达下调,成功鉴定出4种蛋白质即转铁蛋白、锌-a2-糖蛋白、a2-H2-糖蛋白、a1-β-糖蛋白。(3)与PM组比较,PM合并ILD组的7个差异蛋白同时表达下调。成功鉴定出2种蛋白质即β2-糖蛋白1和转铁蛋白。2.第二部分:(1)血清淀粉样蛋白A在PM、PM+ILD、SLE、HD组的血清浓度分别为609.0 ng/L、798.5 ng/L、634.0 ng/L、617.0 ng/L,在各组间比较无显著性差异(P0.05)。(2)凝集素在PM、PM+ILD、SLE、HD组的血清浓度分别为13.0ng/L、14.0 ng/L、11.5 ng/L、8.0 ng/L;与健康对照组比较,PM、PM+ILD、SLE表达上调(差异显著,P0.05),其余组间比较无显著性差异(P0.05)。(3)转铁蛋白在PM、PM+ILD、SLE、HD组的血清浓度分别为4.06 ng/L、4.67 ng/L、3.34 ng/L、3.45 ng/L,在各组间比较无显著性差异(P0.05)。(4)β2-糖蛋白1在PM、PM+ILD、SLE、HD组的血清浓度分别为5.0 ng/L、5.04 ng/L、3.75 ng/L、2.29 ng/L。在各组间比较无显著性差异(P0.05)。【结论】凝集素可作为PM疾病血清生物学标志物的候选分子。
[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 卤16.24y.PM ILD, male 1, female 2, age 47.78 卤19.64y. In the same period, there were 3 healthy persons, 1 male and 2 female, the age was 45.66 卤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 卤15.40 yPM-ILD group 6 cases, age 47.33 卤19.14 ym SLE group age 46.8 卤17.05 yy; healthy control group 20 cases, age 44.76 卤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 卤11 protein spots and 235 卤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.
【学位授予单位】：福建医科大学
【学位级别】：硕士
【学位授予年份】：2015
【分类号】：R593.2;R563

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