氨基酸代谢与缺血性脑中风的相关性研究

Relationship Between Metabolism of Amino Acids and Ischemic Cerebral Stroke in Rats

目的利用基于氯甲酸乙酯衍生化反应的气相色谱-质谱联用技术(GC-MS)的代谢组学测定方法,测定正常组、假手术组和模型组大鼠脑组织中18种氨基酸的含量,利用主成分分析方法(PCA)分析脑内氨基酸代谢与缺血性脑中风的相关性,从代谢组学角度初步揭示缺血性脑中风潜在的生物标志物。方法通过氯甲酸乙酯化反应,将生物样品中的氨基酸、脂肪酸、有机酸、糖类和固醇类物质制成热稳定性强、易挥发的酯或醚,通过优化色谱条件对大鼠脑组织中18种氨基酸含量进行分析测定。结果利用已建立的GC-MS测定方法,测定大鼠脑组织中18种氨基酸的含量,并通过主成分分析方法比较了正常组、假手术组和模型组之间的差异。结论初步发现了13种与缺血性脑中风密切相关的氨基酸类生物标志物,13种化合物分别为丙氨酸、甘氨酸、缬氨酸、谷氨酸、亮氨酸、脯氨酸、天冬氨酸、苯丙氨酸、半胱氨酸、赖氨酸、组氨酸、酪氨酸和色氨酸。

Objective A metabolomics method based on derivation reaction combined with gas chromatography-mass spectrometry was established to investigate 18 amino acids of the endogenous metabolites from the brains of normal rats and modeled rats. The present study was performed to find out the potential biomarkers after analyzing the relationship between metabolism of amino acids and ischemic cerebral stroke in rats by the methods of principal component analysis （PCA）. Methods Amino acids, fatty acids, organic acids, glucose and sterols were transferred into ethers or esters which were heat-stable and highly volatile through silylation derivation reaction. And then the chromatographic conditions were optimized to analyze the cerebral endogenous metabolites of the rats. Results The metabolic fingerprints of the endogenous metabolites were obtained by the optimal method reported in another article. Data of 18 amino acids in the rat brain of normal rats, sham operation rats and model rats was analyzed by PCA. Conclusion Thirteen potential biomarkers of ischemic stroke have found out and they were alanine, glycine, valine, glutamic acid, leucine, proline, aspartic acid, phenylalauine, cysteiue, lysine, histidine, tyrosine, and tryptophane.