HIV感染者口腔代谢物的初步研究

发布时间：2018-04-27 21:38

本文选题：HIV + 口腔　；参考：《昆明医科大学》2017年硕士论文

【摘要】：[目的]口腔对于HIV感染的认识具有重要作用,但因HIV引起的口腔环境改变和口腔病变的发病机制仍不清楚。口腔内的代谢产物主要由宿主和微生物产生,分析口腔代谢物特性,将有助于进一步探明HIV感染相关口腔疾病发病机制及发现新的疾病状态的诊断靶标。目前,仅国外有少量关于HIV感染口腔代谢产物变化的报道,国内尚未开展此领域的研究。本研究拟采用病例对照设计方案,以及高通量的液相色谱-质谱(LC-MS)联用和气相色谱-质谱(GC-MS)联用代谢组学技术,比较健康人群和HIV感染者的口腔代谢物变化,旨在初步探明HIV相关口腔代谢物改变特征,以期为HIV感染早期诊断、病情监测和治疗寻找新的潜在的生物标志物。[方法]拟收集未接受HAART的30例HIV阳性感染者和对照组30例HIV阴性受试者的口腔液标本。分别采用气相色谱-质谱联用和液相色谱-质谱联用方法对全部口腔液样本进行检测及生物信息学分析,最后筛选出有意义的差异代谢物。[结果]30名HIV感染者的CD4+淋巴计数分布在13. 9-200个/μL之间。通过这两种代谢组学方法检测后,先经PCA分析,都证实所有样本主成分相似;再经OPLS-DA分析,证实HIV感染者组的口腔代谢有变化。经过筛选经GC-MS检测发现了十种代谢差异物,其中病例组口腔液葡萄糖、柠檬酸、磷酸盐、D-塔罗糖、半乳糖、赤酮酸内酯和α-氨基己二酸水平升高,而丙二胺、甲氧基吲哚和丁酮酸含量降低;在LC-MS检测中发现了九种代谢差异物,其中病例组口腔液胆碱、L-瓜氨酸、黄嘌呤、2-脱氧核苷、胸苷、磷酰胆碱、丝氨酸-亮氨酸小肽水平升高,而3-羟基-3-甲基戊二酸和L-古洛糖酸γ-内酯水平降低。以这些代谢物质为依据,通过KEGG分析,在GC-MS检测中发现碳水化合物代谢途径和氨基酸代谢途径有变化,而在LC-MS检测中发现氨基酸代谢途径、脂质代谢途径和核苷酸代谢途径受到了影响。最后通过代谢通路分析,在GC-MS检测中发现柠檬酸循环、磷酸盐代谢、缬氨酸、亮氨酸和异亮氨酸的生物合成、赖氨酸的生物合成及β丙氨酸代谢可能受到了影响;LC-MS检测中发现咖啡因代谢、甘油磷脂代谢、精氨酸和脯氨酸代谢、嘌呤代谢和嘧啶代谢可能受到了影响。[结论]用代谢组学方法证实了 HIV感染者的口腔液代谢变化明显,并不能用某一种单一的异常代谢标记物来诊断疾病,但是可以用一组口腔生物标记物反映患者体内代谢变化的轮廓。并且,采用不同的代谢组学方法进行研究可以提供更全面的代谢生物学信息。
[Abstract]:Objective: oral cavity plays an important role in the recognition of HIV infection, but the changes of oral environment and the pathogenesis of oral lesions caused by HIV are still unclear. The metabolites in oral cavity are mainly produced by host and microorganism. Analyzing the characteristics of oral metabolites will be helpful to further study the pathogenesis of oral diseases associated with HIV infection and to find new diagnostic targets of disease status. At present, there are only a few reports about the changes of oral metabolites of HIV infection in foreign countries, but no research in this field has been carried out in China. In this study, a case-control design and high throughput liquid chromatography-mass spectrometry (LC-MS) and gas chromatography-mass spectrometry (GC-MS) metabolic techniques were used to compare the changes of oral metabolites between healthy people and those infected with HIV. The purpose of this study was to identify the characteristics of oral metabolites related to HIV and to find new potential biomarkers for the early diagnosis, disease monitoring and treatment of HIV infection. [methods] Oral fluid samples of 30 HIV positive patients without HAART and 30 HIV negative controls were collected. Gas chromatography-mass spectrometry (GC-MS) and liquid chromatography-mass spectrometry (LC-MS) were used to detect and bioinformatics of all oral liquid samples. [results] the distribution of CD4 lymphatic count in 30 patients with HIV infection was 13.3%. Between 9-200 / 渭 L. After the detection of these two methods, PCA analysis confirmed that all the main components of the samples were similar, and the oral metabolism of HIV infected patients was confirmed by OPLS-DA analysis. Ten metabolic differences were found by screening and GC-MS analysis. The oral fluid glucose, citric acid, phosphate D-taroose, galactose, klycolactone and 伪 -aminoadipic acid increased in the case group, while the level of propanediamine was increased in the case group. The contents of methoxyindole and butanoic acid decreased, nine metabolites were found in LC-MS test, among them, the levels of choline L-citrulline, xanthine 2-deoxynucleoside, thymidine, phosphorylcholine, serine leucine small peptide increased in the case group. However, the levels of 3-hydroxyl-3-methyl glutaric acid and L-glutaric acid 纬-lactone decreased. Based on these metabolites and KEGG analysis, carbohydrate metabolic pathway and amino acid metabolic pathway were found in GC-MS detection, while amino acid metabolic pathway was found in LC-MS assay. Lipid metabolism pathway and nucleotide metabolism pathway are affected. Finally, the citric acid cycle, phosphate metabolism, valine, leucine and isoleucine biosynthesis were found in GC-MS analysis. The biosynthesis of lysine and 尾 -alanine metabolism may be affected by LC-MS. Caffeine metabolism, glycerol phospholipid metabolism, arginine and proline metabolism, purine metabolism and pyrimidine metabolism may be affected. [conclusion] the metabolic changes in oral fluid of HIV infected patients were confirmed by the method of metabonomics, and no single abnormal metabolic marker could be used to diagnose the disease. However, a set of oral biomarkers can be used to reflect the profile of metabolic changes in patients. Moreover, using different metabonomics methods can provide more comprehensive information about metabolic biology.
【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;R781.6

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