基于GC-MS技术的婴儿巨细胞病毒肝炎脾虚湿困证的代谢模式研究

发布时间：2018-02-12 05:36

本文关键词： 婴儿巨细胞病毒肝炎脾虚湿困证代谢组学气相色谱质谱联用　出处：《南京中医药大学》2017年硕士论文　论文类型：学位论文

【摘要】：目的:研究巨细胞病毒(Cytomegalovirus,CMV)肝炎脾虚湿困证患儿血浆和尿液的代谢特征、受干扰的代谢物和代谢通路,探讨CMV肝炎脾虚湿困证的证候实质。方法:选取符合诊断标准的CMV肝炎脾虚湿困证患儿20例设为脾虚湿困组。同时选取同时期同年龄段正常儿童20例设为正常组。采集两组儿童的血浆和尿液,应用气相色谱-三重四级杆质谱仪(gas chromatography-mass spectrometry,GC-MS)检测代谢产物,利用 XCMS软件进行数据提取和数据转化,导入SIMCA-P软件进行多变量分析,并建立主成分分析(Principal Component Analysis,PCA)和正交偏最小二乘法判别分析(Orthogonal Partial Least Squares Discriminant Analysis,OPLS-DA)数学模型,筛选能有效表征婴儿 CMV 肝炎脾虚湿困证的差异性代谢物,并使用MetaboAnalyst分析可能涉及的相关代谢通路来综合评价CMV肝炎脾虚湿困证的证候本质。结果:1.血浆样本代谢组学模式分析结果显示,在所构建的PCA及OPLS-DA模型中正常组血浆样本提取物与婴儿CMV肝炎脾虚湿困证组样本能够良好区分,说明脾虚湿困证组的代谢物特征与正常组具有明显差异。初步鉴定了 22个差异性代谢物作为潜在的生物标记物,其中乳酸、L-丙氨酸、辛酸、L-异亮氨酸、L-脯氨酸、甘氨酸、丝氨酸、L-苏氨酸、氨基丙二酸、蛋氨酸、焦谷氨酸、戊酸、L-谷氨酸、鸟氨酸、L-赖氨酸、L-酪氨酸、肌醇、硬脂酸、L-色氨酸和胆固醇在CMV肝炎脾虚湿困证患儿血浆中呈上调趋势,而3-羟基丁酸、乙二胺四乙酸在CMV肝炎脾虚湿困证患儿血浆中呈下调趋势。2.尿液样本代谢组学模式分析结果显示,在所构建的PCA及OPLS-DA模型中正常组尿液样本提取物与婴儿CMV肝炎脾虚湿困证组样本能够良好区分,说明脾虚湿困证组的代谢物特征与正常组具有明显差异。初步鉴定了 9个差异性代谢物作为潜在的生物标记物,其中甘氨酸、L-苏氨酸、L-赖氨酸、D-甘露醇、D-葡萄糖酸、棕榈酸、尿酸、硬脂酸在CMV肝炎脾虚湿困证患儿尿液中呈上调趋势,而柠檬酸在CMV肝炎脾虚湿困证患儿尿液中呈下调趋势。结论:1.本实验所采用的GC-MS技术能够对CMV肝炎患儿的血浆、尿液进行代谢轮廓分析,并且能够区分和阐释脾虚湿困证组与正常组不同的代谢特征。2.CMV肝炎脾虚湿困证患儿血浆和尿液中主要存在氨基酸代谢及能量代谢紊乱,涉及以下代谢通路,即甘氨酸、丝氨酸和苏氨酸代谢;精氨酸和脯氨酸代谢;丙氨酸、天冬氨酸和谷氨酸代谢;氨酰tRNA合成;赖氨酸降解;丙酮酸代谢;磷酸肌醇代谢;谷氨酰胺和谷氨酸代谢;色氨酸代谢;赖氨酸合成。3.本研究筛选出的差异性代谢物有可能是表征婴儿CMV肝炎脾虚湿困证的潜在的生物标志物。
[Abstract]:Objective: to study the metabolic characteristics of plasma and urine in children with cytomegalovirus cytomegalovirus (CMV) hepatitis due to spleen deficiency and dampness. To explore the syndromes essence of spleen deficiency and dampness in CMV hepatitis. Methods: 20 cases of CMV hepatitis with spleen deficiency dampness were selected as the group of spleen deficiency dampness and 20 cases of normal children of the same age were selected as normal group at the same time. To collect plasma and urine from two groups of children, Gas chromatography-quadruple bar mass spectrometer (chromatography-mass spectrometrymetric GC-MS) was used to detect the metabolites. The data were extracted and transformed by XCMS software, and then imported into SIMCA-P software for multivariate analysis. The mathematical models of principal component analysis (PCA) and orthogonal Partial Least Squares Discriminant analysis (OPLS-DAA) were established. The related metabolic pathways involved in MetaboAnalyst analysis were used to evaluate the syndromes essence of spleen deficiency and dampness in patients with CMV hepatitis. Results: 1.The results of plasma sample metabonomics analysis showed that, In the constructed PCA and OPLS-DA model, the samples of normal group and infant group of CMV hepatitis with spleen deficiency and dampness could be distinguished well. 22 different metabolites were identified as potential biomarkers, including lactic acid L-alanine, octanoic acid L-isoleucine, L-proline, glycine. Serine L-threonine, aminomalonic acid, methionine, pyroglutamic acid, valerate L-glutamic acid, ornithine L-lysine L-tyrosine, inositol, stearic acid L-tryptophan and cholesterol showed an upward trend in the plasma of children with CMV hepatitis due to spleen deficiency. However, the plasma levels of 3-hydroxybutyric acid and ethylenediamine tetraacetic acid were down-regulated in children with CMV hepatitis due to spleen deficiency and dampness. In the constructed PCA and OPLS-DA model, the urine samples of normal group could be distinguished from those of CMV hepatitis group with dampness and dampness syndrome of spleen deficiency in infantile hepatitis. 9 different metabolites were identified as potential biomarkers, among them, glycine L-threonine, L-lysine, D-mannitol, D-gluconic acid and palmitic acid. Uric acid and stearic acid were up-regulated in the urine of children with spleen deficiency and dampness of CMV hepatitis, while citric acid was down-regulated in the urine of children with CMV hepatitis. Conclusion: 1. The GC-MS technique used in this study can be used to treat the plasma of children with CMV hepatitis. Urine metabolism profile analysis, and can distinguish and explain the spleen deficiency dampness trapped syndrome group and normal group of different metabolic characteristics. 2. CMV hepatitis spleen deficiency dampness trapped children plasma and urine mainly exist amino acid metabolism and energy metabolism disorders. Related to the following metabolic pathways: glycine, serine and threonine metabolism; arginine and proline metabolism; alanine, aspartic acid and glutamate metabolism; tRNA synthesis; lysine degradation; pyruvate metabolism; inositol phosphate metabolism; Glutamine and glutamate metabolism; tryptophan metabolism; lysine synthesis. The differential metabolites screened in this study may be a potential biomarker to characterize spleen deficiency and dampness in infants with CMV hepatitis.
【学位授予单位】：南京中医药大学
【学位级别】：硕士
【学位授予年份】：2017
【分类号】：R272

【参考文献】

中国期刊全文数据库前10条

1 崔扬;郭慧;匡海学;王秋红;;代谢组学技术在中医证型研究中的应用概述[J];中华中医药杂志;2017年02期

2 赵亚飞;张一颖;谢雁鸣;支英杰;赵维;庄严;;儿童使用双环醇片联合甘草酸注射剂的临床用药特征分析[J];中华中医药杂志;2017年02期

3 谭艳芳;欧阳文献;姜涛;袁鹤立;邓惠;李双杰;;槐杞黄颗粒治疗巨细胞病毒肝炎疗效观察及对CD4~+T细胞相关因子表达的影响[J];中南药学;2016年12期

4 崔孟颖;柳洁;杨蕾;Cathie Martin;陈晓亚;赵清;;黄芩,由古籍里走进现代生活的药用植物[J];世界科学技术-中医药现代化;2016年11期

5 穆静;杨燕;刘畅;;中药治疗婴儿巨细胞病毒性肝炎(湿热内蕴证)40例临床观察[J];中国中医急症;2016年10期

6 豆玉凤;史艳平;;凉血退黄汤治疗婴儿巨细胞病毒肝炎疗效观察[J];陕西中医;2016年10期

7 张旭;曹美玲;;茵栀黄对早产儿巨细胞病毒肝炎的临床疗效研究[J];中国医科大学学报;2016年10期

8 王芳;鄢素琪;邓玉萍;万琦;李红;;利胆合剂对肝病综合征患儿的临床疗效及其对炎症因子的影响[J];中国临床药理学杂志;2016年06期

9 吴晓波;吕贞燕;何勇;;更昔洛韦联合丙种免疫球蛋白治疗婴儿巨细胞病毒感染的系统评价[J];临床药物治疗杂志;2016年02期

10 高明;任章平;;熊去氧胆酸联合护肝药物对婴儿肝炎综合征生化指标的影响[J];肝脏;2016年02期