重型肝炎预后相关蛋白质质谱分析及预测模型构建

发布时间：2018-06-15 21:18

本文选题：重型肝炎 + 预后　；参考：《西南医科大学》2017年硕士论文

【摘要】：目的:本研究旨在通过应用SELDI-TOF-MS(surface enhanced laser desorption/ionization time-of flight mass spectrometry)技术获得HBV相关重型肝炎不同临床结局患者和正常人血浆中的蛋白质指纹图谱,筛选出与重型肝炎患者预后相关的差异蛋白,并利用这些差异蛋白初步构建重型肝炎的预后预测模型,为重型肝炎预后判断提供可靠的、敏感的指标。方法:收集西南医科大学附属医院感染科住院部HBV相关重型肝炎患者35例和我院体检中心健康志愿者15例的血浆标本,分别作为实验组和对照组。同时收集患者的人口学资料和临床资料(包括肝功能、凝血功能等)。实验组患者随访时间至少12周。根据随访患者病情的转归不同分为死亡组和存活组。应用SELDI-TOF-MS蛋白质芯片技术检测HBV相关重型肝炎患者和正常健康人血浆样本,获得弱阳离子交换芯片CM10的蛋白质指纹图谱。使用ProteinChip 3.2软件和Biomarker Wizard软件识别蛋白指纹图谱,并利用SPSS 16.0软件对所采集的数据进行统计学分析,利用Biomarker Patterns(BPS 5.0)软件采用树形分类方法对HBV相关重型肝炎患者初步建立预后预测模型,并计算该模型对HBV相关重型肝炎患者预后判断的敏感性和特异性。计量资料采用均数±标准差(x±s)描述。组间比较采用两个独立样本t检验。以P0.05示有统计学意义。结果:CM10蛋白芯片检测结果显示,HBV相关重型肝炎患者和健康对照组血浆样本在蛋白分子量1000Da-50000Da的范围内一共检测出57个蛋白峰表达水平有差异,其中与健康对照组相比较,重型肝炎存活组有25个蛋白峰表达有显著差异(p0.05),其中有6个蛋白峰显著升高,它们是m2017_57、m7568_86、m7937_87、m15109_0、m15268_2、m15845_0;有19个蛋白峰显著降低,它们是m2196_71、m2235_78、m2747_23、m2796_85、m2881_48、m2913_26、m3224_17、m3322_84、m4309_41、m6437_01、m6634_84、m6839_64、m8601_12、m9185_56、m28057_7、m33395_2、m34404_9、m44842_1、m45146_8。重型肝炎死亡组有24个蛋白峰表达有显著差异(p0.05),其中有6个蛋白峰显著升高,它们是m2017_57、m6634_84、m7568_86、m7937_87、m15268_2、m15845_0;有18个蛋白峰显著降低,它们是m2235_78、m2747_23、m2881_48、m2913_26、m3224_17、m3322_84、m3376_49、m4309_41、m5908_26、m6437_01、m6584_63、m6839_64、m8601_12、m14670_8、m15109_0、m28057_7、m33395_2、m34404_9。重型肝炎存活组和死亡组比较发现有16个蛋白峰表达有显著差异(p0.05),它们分别是m3376_49、m3479_65、m3492_79、m3936_56、m5649_11、m7568_86、m7766_46、m7937_87、m8601_12、m9290_44、m11692_6、m14670_8、m15109_0、m15268_2、m15845_0、m45146_8。应用biomarkerpatterns软件对重型肝炎存活组和死亡组之间有统计学意义的蛋白峰进行比较分析,并最终筛选出5个差异蛋白峰组成树形决策模型。这5个差异蛋白峰分别是m3492_79、m5649_11、m3936_56、m7568_86和m8601_12,所建立的模型对重型肝炎预后判断的敏感性为72.00%,特异性为100.00%。结论:1.应用seldi-tof-ms技术可以发现hbv相关重型肝炎发生、发展过程中某些蛋白质表达水平有差异,不同临床结局的重型肝炎患者血浆蛋白峰表达差异比较显著。2.利用这些差异蛋白峰构建的预后预测模型对HBV相关重型肝炎患者的临床结局判断有着重要的意义,并提示这些差异表达的蛋白质可能和HBV相关重型肝炎预后关系密切,但尚需进一步研究证实。
[Abstract]:Objective: the purpose of this study was to obtain the protein fingerprints in the plasma of patients with different clinical outcomes of severe hepatitis and normal human plasma by using SELDI-TOF-MS (surface enhanced laser desorption/ionization time-of flight mass spectrometry) technology, and to screen out the differential proteins associated with the prognosis of patients with severe hepatitis and use this method. Some differential proteins were initially constructed to predict the prognosis of severe hepatitis and provide a reliable and sensitive indicator for the prognosis of severe hepatitis. Methods: 35 cases of severe hepatitis in the hospital department of infection department of the Affiliated Hospital of Southwest Medical University were collected in 35 cases of severe hepatitis patients and 15 cases of healthy volunteers in the medical center of our hospital. The patients' demographic and clinical data (including liver function, coagulation function, etc.) were collected at the same time. The patients in the experimental group were followed up for at least 12 weeks. According to the prognosis of the patients, the patients were divided into the death group and the survival group according to the prognosis of the patients. The SELDI-TOF-MS protein chip technique was used to detect the plasma samples of patients with severe hepatitis HBV and normal health. The protein fingerprint of the weak cation exchange chip CM10 was obtained. The ProteinChip 3.2 software and Biomarker Wizard software were used to identify the protein fingerprints, and the data collected by the SPSS 16 software were statistically analyzed. The Biomarker Patterns (BPS 5) soft parts were used to use the tree classification method for the severe hepatitis associated with HBV. The prognosis prediction model was preliminarily established, and the sensitivity and specificity of the model to the prognosis of patients with HBV related severe hepatitis were calculated. The measurement data were described with mean number + standard deviation (x + s). Two independent sample t tests were used among the groups. The results of P0.05 were statistically significant. Results: the results of CM10 protein chip detection showed that HBV related heavy duty was heavy. The plasma samples of the hepatitis patients and the healthy control group detected 57 protein peaks in the range of protein molecular weight 1000Da-50000Da. Compared with the healthy control group, there were 25 protein peaks in the survival group of severe hepatitis (P0.05), of which 6 protein peaks were significantly increased, they were m2017_57, m7568_86 M7937_87, m15109_0, m15268_2, m15845_0, which have 19 protein peaks, which are m2196_71, m2235_78, m2747_23, m2796_85, m2881_48, m2913_26, m3224_17, there are 24 protein peaks in the severe hepatitis death group. There are 6 protein peaks in P0.05, which are m2017_57, m6634_84, m7568_86, m7937_87, m15268_2, m15845_0, and there are 18 protein peaks, which are m2235_78, m2747_23, m2881_48, m2913_26. It is found that there are 16 protein peaks in the survival group and the death group of the severe hepatitis m34404_9. (P0.05). They are m3376_49, m3479_65, m3492_79, m3936_56, m5649_11, m7568_86, m7766_46, m7937_87. A statistically significant protein peak was compared between the survival group of severe hepatitis and the death group, and 5 differential protein peaks were selected to form a tree decision model. The 5 differential protein peaks were m3492_79, m5649_11, m3936_56, m7568_86 and m8601_12, and the sensitivity of the established model to the prognosis of severe hepatitis was 72%. The opposite sex is 100.00%. conclusion: 1. SELDI-TOF-MS technique can be used to detect the occurrence of severe hepatitis associated with HBV. There is a difference in the expression level of some proteins in the course of development. The difference of the expression of protein peak in the plasma of severe hepatitis patients with different clinical outcomes is more significant than that of.2., and the prognosis prediction model constructed by these differential protein peaks for HBV related severe liver disease It is important to judge the clinical outcome of inflammatory patients and suggest that these differentially expressed proteins may be closely related to the prognosis of HBV related severe hepatitis, but further research is needed.
【学位授予单位】：西南医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R512.62

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