HIV感染者口腔代谢物变化与CD4水平的关系

发布时间：2018-08-02 20:51

【摘要】：[目的]口腔代谢产物的改变可用于疾病状态和/或治疗耐受的诊断标志,因此,分析口腔代谢物特征有助于发现HIV疾病进程的预测靶标。迄今,尚鲜见HIV感染者口腔代谢物分布特征与免疫学重要指标CD4计数水平关系的研究,本研究拟使用GC-TOF-MS检测平台,对云南地区部分HIV感染者口腔液代谢物进行检测,并分析其与CD4变化关系,以期发现与HIV感染疾病进程有关的新的标志物。[方法]选取2016年9月至2016年12月到昆明市第三人民医院感染一科就诊且尚未接受高效抗逆转录病毒治疗(HAART)的HIV感染者为病例组,收集受试者的基本信息和口腔液样本,并记录CD4计数。口腔液标本采用GC-TOF-MS进行非靶标代谢组学检测,对所得数据进行预处理,然后用主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLS-DA)、代谢物定性及差异代谢物筛选、代谢通路分析和层次聚类分析等方法,比较不同CD4水平的HIV感染者之间口腔代谢物变化。[结果]本研究共收集到90例HIV感染者的口腔液标本90份,按外周血CD4计数水平分为A组(CD4计数200个/mm3)、B组(CD4计数200-500个/mm3)和C组(CD4计数500个/mm3),每组各30例。所获标本经GC-TOF-MS检测及生物信息学分析,主要结果如下:(1) PCA分析结果可看出:A组和B组及A组和C组的不同样品之间大多排列紧凑、聚合成簇,显示出实验较好的重复性,B组和C组的不同样品之间大多排列比较分散,未能准确的显示出实验较好的重复性;不同实验分组A组和B组之间,A组和C组之间则互相分离,可以做出清晰的划分,显示出不错的特异性,不同实验分组B组和C组的样本出现交叠现象,表明部分样本间相似度较高,差异性较小。(2) OPLS-DA分析结果可看出:A和B两组样本及A和C两组样本互相分离,可以做出清晰的划分,可以说两组样本区分非常显著,样本基本全部处于95%置信区间内,表明此数据值得进一步研究。而B和C两组样本区分不是非常显著,但是可以做出划分,样本基本全部处于95%置信区间内,表明此数据也值得进一步研究。(3)代谢物定性及差异代谢物筛选、代谢通路分析和层次聚类分析结果可看出:A组和B组筛选出来的差异代谢物共116种,差异代谢物映射的KEGG通路7条,其中,最主要的差异代谢通路分别是半乳糖代谢,磷酸戊糖,嘧啶代谢、亚油酸代谢、果糖和甘露糖代谢。A组和C组筛选出来的差异代谢物为133种,差异代谢物映射的KEGG通路9条,其中,最主要的差异代谢通路分别是半乳糖代谢、磷酸戊糖、嘌吟代谢、嘧啶代谢、亚油酸代谢、果糖和甘露糖代谢。B组和C组筛选出来的差异代谢物相对较少,共34种,差异代谢物映射的KEGG通路4条,主要的代谢通路分别是半乳糖代谢、果糖和甘露糖代谢和嘌呤代谢。[结论]本研究应用先进的GC-TOF-MS检测平台,分析了外周血不同CD4计数水平的HIV感染者口腔代谢物的变化特征。研究发现,CD4计数水平不同,患者口腔内代谢差异明显程度也不同。研究成果可以为发现HIV感染者新的标志物及疾病控制提供了新的科学依据。
[Abstract]:[Objective] changes in oral metabolites can be used in the diagnosis of disease status and / or treatment tolerance. Therefore, the analysis of oral metabolites is helpful for the detection of the target of the HIV disease process. So far, the study of the relationship between the distribution characteristics of oral metabolites and the level of the important immunological index of CD4 counts in HIV infected people. This study is to be used in this study. GC-TOF-MS detection platform was used to detect the oral liquid metabolites of some HIV infected people in Yunnan, and analyze the relationship between them and CD4, in order to find new markers related to the process of HIV infection. [Methods] select the third people's Hospital from September 2016 to December 2016 to be infected with one family and have not received high efficiency antiretroviral The patients with HIV infection (HAART) were the case group. The basic information and oral liquid samples were collected and the CD4 counts were recorded. The oral liquid specimens were detected by GC-TOF-MS for non target metabolism, preprocessed the data, and then used principal component analysis (PCA), orthogonal partial least square method discriminant analysis (OPLS-DA) and metabolite. Sexual and differential metabolites screening, metabolic pathway analysis and hierarchical cluster analysis were used to compare the changes in oral metabolites between HIV infected persons at different CD4 levels. [results] 90 specimens of oral liquid were collected from 90 cases of HIV infection, and were divided into A group (CD4 count 200 /mm3) and B group (CD4 count 200-500 /mm3) according to the CD4 count level of peripheral blood. And group C (CD4 count 500 /mm3), 30 cases in each group. The samples were detected by GC-TOF-MS and bioinformatics analysis. The main results were as follows: (1) the results of PCA analysis showed that the different samples between A and B groups and A and C groups were mostly arranged and clustered, showing good reproducibility, most of the different samples between B and C groups The arrangement of the A group and the B group, the A group and the C group, separated each other, can make a clear division and show the good specificity. The samples of the B and C groups in the different experimental groups appear to overlap, indicating that the similarity of some samples is higher and the difference is more different. (2) (2) the results of OPLS-DA analysis show that two samples of A and B and two groups of A and C can be separated from each other and can make a clear division. It can be said that the two groups of samples are very distinct, and the sample is basically in the 95% confidence interval, indicating that this data is worth further study. And the distinction between the B and the C two groups is not very significant, but can be made. The sample was basically in the 95% confidence interval, indicating that this data was also worthy of further study. (3) the metabolite qualitative and differential metabolite screening, metabolic pathway analysis and hierarchical cluster analysis can be seen: A and B groups of different metabolites selected 116 species, differential metabolite mapping of the KEGG pathway 7, among them, the most important The differential metabolic pathways were galactose metabolism, pentose phosphate, pyrimidine metabolism, linoleic acid metabolism,.A group and C group of fructose and mannose metabolism, 133 different metabolites, and 9 KEGG pathway of differential metabolite mapping, among which the main differential metabolic pathways were galactose metabolism, pentose phosphate, purinyin metabolism, pyrimidine generation. Metabolites of linoleic acid, linoleic acid, fructose and mannose metabolized in group.B and C group were relatively few, 34 species, 4 KEGG pathways of differential metabolite mapping, the main metabolic pathways were galactose metabolism, fructose and mannose metabolism and purine metabolism. [Conclusion] this study applied advanced GC-TOF-MS detection platform and analyzed outside The changes in oral metabolites of HIV infected people with different CD4 counting levels in the peripheral blood have been found to be different in different levels of CD4 count, and the research results can provide new scientific evidence for the discovery of new markers and disease control of HIV infected people.
【学位授予单位】：昆明医科大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R512.91

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