基于代谢组学分析缺血性中风痰湿证生物标志物特点研究

Study on Biomarkers of Phlegm-dampness Syndrome in Ischemic Stroke Based on Metabonomics

【目的】运用核磁共振代谢组学方法研究缺血性中风(IS)痰湿证患者和健康人群血清代谢产物差异,总结IS痰湿证相关生物标志物特点,探讨疾病发生机制。【方法】选取13例明确诊断为IS、中医证型为痰湿证的患者作为病例组(IS痰湿证组),另选取18例中医体质为平和质的健康体检者作为正常对照组(平和质组)。对收集的IS痰湿证患者和平和质人群的血清样本进行核磁共振氢谱处理,使用SIMCA软件进行建模分析,经主成分分析(PCA)、正交偏最小二乘法-判别分析(OPLS-DA)及置换检验,最终得到2组间的差异代谢物。【结果】IS痰湿证患者的差异代谢物分别为1-甲基组氨酸、醋酸、丙氨酸、丙酮、柠檬酸、肌酸、乙醇胺、甲酸、谷氨酸、谷氨酰胺、甘氨酸、异丁酸、异亮氨酸、乳酸、低密度脂蛋白、亮氨酸、脂质、赖氨酸、甲醇、蛋氨酸、肌醇、N-乙酰糖蛋白、苯丙氨酸、酪氨酸、缬氨酸、极低密度脂蛋白、α-葡萄糖、β-葡萄糖,与平和质组比较,差异均有统计学意义(P<0.05)。【结论】以上代谢差异物均为IS痰湿证潜在生物标志物;IS痰湿证血清代谢组学变化机制与氨基酸代谢、能量代谢、脂类代谢密切相关,具体影响涉及血脑屏障破坏、三羧酸循环无氧糖酵解增强、叶酸代谢通路障碍、胆碱代谢异常。

Objective The differences of serum metabolites between ischemic stroke(IS)patients with phlegm-dampness syndrome and healthy volunteers were investigated by using nuclear magnetic resonance(NMR)metabonomics,so as to summarize the characteristics of related biomarkers of IS patients with phlegm-dampness syndrome,and to explore the pathogenesis of the disease.Methods Thirteen patients confirmed as IS and differentiated as phlegm-dampness syndrome were used as the case study group,and 18 healthy volunteers having physical examination and differentiated as balanced constitution served as the normal control group.Serum samples of the two groups were treated with 1H NMR,and then the data was processed by SIMCA modeling analysis,principal component analysis(PCA),orthogonal partial least squares-discriminant analysis(OPLS-DA)and permutation test for the achievement of the differential metabolites from the two groups.Results The differential metabolites of IS patients with phlegm-dampness syndrome were 1-methylhistidine,acetic acid,alanine,acetone,citric acid,creatine,ethanolamine,formic acid,glutamic acid,glutamine,glycine,isobutyric acid,isoleucine,lactic acid,low-density lipoprotein,leucine,lipids,lysine,methanol,methionine,inositol,N-acetyl glycoprotein,phenylalanine,tyrosine,valine,very low density lipoprotein,alpha-glucose and betaglucose,which differed from those in the normal control group(P<0.05).Conclusion The above differential metabolites are potential biomarkers of IS patients with phlegm-dampness syndrome.The mechanism of changes in serum metabolomics of phlegm-dampness type of IS syndrome is closely related to amino acid metabolism,energy metabolism and lipid metabolism,and the specific mechanisms involve the destruction of blood-brain barrier,the enhancement of anaerobic glycolysis of tricarboxylic acid cycle,the disorder of folic acid metabolic pathway and the abnormal metabolism of choline.