基于代谢组学平台的胶质瘤患者血浆代谢标志物研究

发布时间：2018-09-12 13:41

【摘要】：目的:胶质瘤是中枢神经系统最常见的恶性肿瘤,具有发病率、复发率、死亡率高等特点。胶质瘤发病原因和发展机制均不明确,因此临床诊疗效果较差。胶质瘤发生与发展伴随着细胞代谢的改变。细胞代谢产物处于基因和蛋白质的下游,是基因和蛋白质生理、生化功能变化的最终反映。胶质瘤的发生与发展伴随着细胞代谢产物种类及含量的变化,用代谢组学技术检测与分析这些代谢物变化的机制、从代谢层面考察胶质瘤细胞恶性进展,将有助于阐明胶质瘤恶性进展的分子机制,对探索靶向代谢途径治疗胶质瘤提供新的思路。方法:1)通过医学检查确定为健康的健康志愿者70例,2012年6月至2013年3月新疆医科大学第一附属医院收治的胶质瘤患者85例:其中手术前患者70例、手术后患者15例。患者平均年龄64.2岁,年龄范围为46至77岁。对血浆样品进行1Hnm R测试与分析;探讨胶质瘤发病过程的血浆代谢物的变化。2)通过体检确定为身体健康的志愿者50名;2012年6月至2013年3月新疆医科大学第一附属医院收治的胶质瘤患者70例;患者平均年龄64.2岁,年龄范围为46至77岁。收集健康志愿者和患者术前清晨空腹血液并收集血浆,-80℃保存至测试。建立高效液相色谱(HPLC)分析胶质瘤血浆样品中游离氨基酸的代谢靶标分析技术,以胶质瘤患者血浆样本为研究对象,测定氨基酸代谢谱,结合多元统计分析评估氨基酸代谢谱在胶质瘤发病过程的变化情况。结果:1)我们对健康志愿者与胶质瘤手术前患者1H-NMR图谱的分析研究发现了20种代谢产物在两组中有明显的差异。这些代谢物主要参与体内关键的代谢途径,包括糖酵解,氨基酸代谢,TCA循环和脂肪酸代谢等。与健康对照组相比,胶质瘤患者血浆中异亮氨酸、亮氨酸、缬氨酸、丙氨酸、酪氨酸、苯丙氨酸、1-甲基组氨酸、糖蛋白、谷氨酸、肌醇、柠檬酸盐、肌酐、胆碱、α-葡萄糖、β-葡萄糖和乳酸(P0.05)等代谢物含量明显降低;而极低密度脂蛋白、低密度脂蛋白、不饱和脂肪和丙酮酸(P0.05)等含量明显升高。对胶质瘤手术前与手术后患者1H-NMR图谱的分析研究发现了9种代谢产物在两组中有明显差异。胶质瘤患者手术后血浆异亮氨酸、亮氨酸、缬氨酸、乳酸、谷氨酸、肌醇、酪氨酸和苯丙氨酸等代谢物含量明显升高(P0.05);而丙氨酸含量明显降低(P0.05)。2)健康对照组与胶质瘤患者血浆氨基酸含量进行比较除脯氨酸含量没有发生变化以外,其他氨基酸均发生了明显的变化。与健康志愿者比较,胶质瘤患者血浆中的天冬氨酸、谷氨酸、丝氨酸、精氨酸、苏氨酸、丙氨酸、缬氨酸、蛋氨酸、异亮氨酸、亮氨酸和苯丙氨酸等11种氨基酸含量明显下降,差异有统计学意义(P0.05);但是,甘氨酸、赖氨酸、胱氨酸、酪氨酸和组氨酸含量明显升高,差异有统计学意义(P0.05)。结论:本研究结果显示,核磁共振为基础的代谢组学研究方法把血浆1H-NMR数据与多元统计分析方法相结合,发现了胶质瘤患者体内所发生的一系列代谢变化,包括糖代谢、脂肪代谢及三羧酸循环过程中的代谢。研究结果不仅有利于胶质瘤的诊断在分子水平上有一定的提高提供一种机会和可能性也有利于对胶质瘤发生、发展过程的进一步研究提供可能。代谢组学研究对这一类肿瘤的早起发现和预后评价体系的建立提供一种新的研究思路。同时本研究采用HPLC技术测试胶质瘤患者血浆氨基酸代谢谱的变化规律。通过分析确定了胶质瘤患者氨基酸代谢谱的变化规律及其机制。研究发现,胶质瘤患者体内氨基酸代谢存在明显的紊乱,胶质瘤患者血浆中谷氨酸、丙氨酸、缬氨酸、赖氨酸、胱氨酸、酪氨酸和甘氨酸等氨基酸可以作为胶质瘤潜在的氨基酸代谢标志物;其中谷氨酸、丙氨酸和缬氨酸等氨基酸与胶质瘤的发生成负相关,而赖氨酸、胱氨酸、酪氨酸和甘氨酸等氨基酸与胶质瘤的发生成正相关。
[Abstract]:Objective: Glioma is the most common malignant tumor of the central nervous system, with the characteristics of high incidence, recurrence rate and mortality. The pathogenesis and development mechanism of glioma are not clear, so the clinical diagnosis and treatment effect is poor. The occurrence and development of gliomas are accompanied by changes in the types and contents of cell metabolites. Metabonomic techniques are used to detect and analyze the mechanisms of these metabolites and to investigate the malignant progress of gliomas from the metabolic level. This will help to clarify the malignant progress of gliomas. Methods: 1) 70 healthy volunteers were identified by medical examination, and 85 patients with glioma were admitted to the First Affiliated Hospital of Xinjiang Medical University from June 2012 to March 2013. Among them, 70 were pre-operative and 15 were post-operative. 2 years old, ranging from 46 to 77 years old. The plasma samples were tested and analyzed by 1Hnm R. The changes of plasma metabolites during the onset of glioma were investigated. 2) 50 healthy volunteers were identified by physical examination; 70 patients with glioma were admitted to the First Affiliated Hospital of Xinjiang Medical University from June 2012 to March 2013; the average age of the patients was 64.2 years old. A high performance liquid chromatographic (HPLC) method was developed for the determination of free amino acids in plasma samples of gliomas. Results: 1) Our analysis of 1H-NMR profiles of healthy volunteers and pre-operative glioma patients revealed significant differences in 20 metabolites between the two groups. Amino acid metabolism, TCA cycle and fatty acid metabolism. Compared with healthy controls, plasma levels of isoleucine, leucine, valine, alanine, tyrosine, phenylalanine, 1-methylhistidine, glycoprotein, glutamic acid, inositol, citrate, creatinine, choline, alpha-glucose, beta-glucose and lactic acid (P 0.05) were significantly higher in glioma patients. The levels of VLDL, LDL, unsaturated fat and pyruvic acid (P 0.05) were significantly increased. The 1H-NMR profiles of glioma patients before and after surgery showed that there were significant differences in 9 metabolites between the two groups. The contents of amino acids in plasma of healthy control group and glioma patients were significantly higher than those of healthy volunteers except proline content. The contents of 11 amino acids in plasma of tumor patients were significantly decreased (P 0.05), but the contents of glycine, lysine, cystine, tyrosine and histidine were significantly increased. CONCLUSIONS: The results of this study suggest that the 1H-NMR data of plasma combined with multivariate statistical analysis can reveal a series of metabolic changes in glioma patients, including glucose metabolism, lipid metabolism and metabolism during tricarboxylic acid cycle. Metabonomics provides a new approach to the early detection of gliomas and the establishment of a prognostic evaluation system. The changes of plasma amino acid metabolism in glioma patients were determined by HPLC. The changes of amino acid metabolism in glioma patients were analyzed and the mechanism was determined. Amino acids such as amino acid, tyrosine and glycine can be used as potential metabolic markers of amino acids in gliomas. Amino acids such as glutamate, alanine and valine are negatively correlated with the occurrence of gliomas, while amino acids such as lysine, cystine, tyrosine and glycine are positively correlated with the occurrence of gliomas.
【学位授予单位】：新疆医科大学
【学位级别】：博士
【学位授予年份】：2016
【分类号】：R739.41

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