丙型肝炎病毒自发清除者的血清代谢组学研究

发布时间：2018-02-06 05:25

本文关键词： 丙型肝炎病毒自发清除丙型肝炎慢性感染血清代谢组学液相色谱-串联质谱联用（LC-MS/MS）主成分分析（PCA）正交偏最小二乘法-判别分析（OPLS-DA）　出处：《吉林大学》2013年硕士论文　论文类型：学位论文

【摘要】：丙型肝炎是一种严重威胁全球公共健康的疾病，目前尚无有效疫苗可预防。据估计全世界范围内约有1.8亿人感染HCV，主要通过输血和血制品、破损的皮肤和黏膜等途径传播。HCV急性感染者中约有20%～30%可实现病毒自发清除，感染HCV6个月后病毒仍未清除称为慢性丙型肝炎，约50%～80%的HCV感染者将进展为慢性感染状态，其中20%～30%的患者将发展为肝硬化或肝癌。目前丙肝的标准治疗方法为聚乙二醇干扰素联合利巴韦林(PEG-IFN-α/RBV)治疗，由于疗程长、不良反应大、IFN价格相对较高和持续病毒学应答率较低等原因，给患者治疗带来较大负担。代谢组学研究源于代谢轮廓分析（metabolic profiling），作为一个新的研究手段，已成为系统生物学的重要组成部分，主要反映在外界刺激下由于组织细胞的病理生理学改变而引起的代谢产物相应改变。代谢组学多以相对分子量小于1000Da的内源性代谢产物为研究对象，，可从整体上发现生物样本代谢组分的改变。目前在疾病诊断与治疗、新药研制开发、药物作用和毒性机制研究等方面已显示出较明显优势。本研究纳入了丙型肝炎病毒自发清除组、慢性丙型肝炎组和健康对照组各30例样本作为研究对象，采用快速液相色谱-串联质谱联用（LC-MS/MS）技术，应用主成分分析（PCA）、正交偏最小二乘法-判别分析（OPLS-DA）进行模式识别，然后通过变量重要性因子（VIP）、非参数检验，结合数据库检索筛选鉴定出慢性丙型肝炎组、健康对照组及丙型肝炎病毒自发清除组间差异的代谢物。研究共发现25种代谢物在以上三组中存在显著差异，其中7个变量被鉴定为花生四烯酸、棕榈油酸、葵酰基肉碱、溶血磷脂酰胆碱（20:5,16:0）、溶血磷脂酰乙醇胺（16:0,18:0），涉及脂肪酸、磷脂等代谢。其中花生四烯酸以及未鉴定出明确结构的m/z179.0719、m/z382.1360、m/z548.3475、m/z680.4281、m/z303.2323等物质与丙型肝炎病毒自发清除组的相关性较好，受试者工作特征曲线（ROC）下面积为0.887~0.977，具有较好的特异性和敏感性。丙型肝炎病毒自发清除组、慢性丙型肝炎组和健康对照组在血清代谢水平上存在较明显差异，这些差异物可能与丙型肝炎病毒感染后出现的不同临床转归有关。
[Abstract]:Hepatitis C is a serious threat to global public health, and there is no effective vaccine to prevent it. It is estimated that about 180 million people worldwide are infected with HCV, mainly through blood transfusions and blood products. About 20% of the acute infected people with HCV6 can realize the spontaneous clearance of the virus, and the virus has not been cleared of chronic hepatitis C for months after infection with HCV6. About 50% of HCV infected people will progress to chronic infection. Twenty percent of the patients will develop cirrhosis or liver cancer. The current standard treatment for hepatitis C is PEG-IFN- 伪 / RBV combined with pebavirin. Because of the long course of treatment, the high price of IFNs and the low rate of persistent virological response, the treatment of IFNs brings a great burden to the patients. Metabonomics, as a new research tool, has become an important part of system biology because it originated from metabolic profile analysis. It is mainly reflected in the corresponding changes of metabolites caused by pathophysiological changes of histocytes under external stimulation. Metabolites with relative molecular weight less than 1000Da are the research objects. The changes of metabolic components in biological samples can be found as a whole. At present, there are obvious advantages in the diagnosis and treatment of diseases, the development of new drugs, the study of drug action and the mechanism of toxicity, etc. In this study, 30 samples of hepatitis C virus spontaneous clearance group, 30 cases of chronic hepatitis C group and 30 healthy control group were included as the study objects. A rapid liquid chromatography-tandem mass spectrometry (LC-MS / MS) technique was used to identify the patterns by using principal component analysis (PCA) and orthogonal partial least-squares discriminant analysis (OPLS-DA). Then the metabolites of chronic hepatitis C group, healthy control group and hepatitis C virus spontaneous clearance group were identified by variable importance factor (VIPP), nonparametric test and database retrieval. A total of 25 metabolites were identified as arachidonic acid, palmitic oil acid, sunacylcarnitine and lysophosphatidylcholine 20: 5, among which 7 variables were identified as arachidonic acid, palmitic oil acid and lysophosphatidylcholine 20: 5. 16: 0, lysophosphatidyl ethanolamine 16: 0 0: 0, involved in fatty acid, phospholipid metabolism, among them arachidonic acid and mrz179.0719, whose structure has not been identified. M / z 382.1360 m / m / z 548.3475N / m / z 680.4281m / z 303.2323 and so on, have a good correlation with the group of spontaneous clearance of hepatitis C virus. The area under the operating characteristic curve (ROC) is 0. 887 ~ 0. 977, which has good specificity and sensitivity. There were significant differences in serum metabolic level among the spontaneous clearance group of hepatitis C virus, chronic hepatitis C group and healthy control group. These differences may be related to different clinical outcomes after hepatitis C virus infection.
【学位授予单位】：吉林大学
【学位级别】：硕士
【学位授予年份】：2013
【分类号】：R512.63

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