冠心病心血瘀阻证血浆代谢组学的检测分析

目的寻找冠心病(coronary heart disease,CHD)心血瘀阻证患者血浆代谢产物谱,分析CHD心血瘀阻证血浆层面的代谢途径。方法采用QP2010气相色谱-质谱联用仪(GC-MS)检测CHD心血瘀阻证组和CHD非心血瘀阻证组及健康对照组血浆代谢产物,对不同组之间代谢产物含量的变化主成分分析(principal components analysis,PCA)、偏最小二乘法分析(partial lease squares,PLS)。结果 (1)主成分分析:显示3组样本主成分积分值,3组分聚于椭圆形散点图的3个区域,其中CHD心血瘀阻证组中乳酸、β-羟基丁酸、尿素、油酸、硬脂酸、花生四烯酸物质含量上调,而柠檬酸含量下调。(2)偏最小二乘法分析:显示健康组远离CHD心血瘀阻证组与CHD非心血瘀阻证组,CHD心血瘀阻证组与CHD非心血瘀阻证组位置比较靠近,可以判别他们同属一类;标准化偏最小二乘系数图显示CHD心血瘀阻证组与硬脂酸、花生四烯酸、尿素、乳酸、β-羟基丁酸呈正相关,与油酸、脯氨酸、甘氨酸、柠檬酸呈负相关。依据VIP值排列各变量与药物干预的密切程度,依次是花生四烯酸、硬脂酸、乳酸、尿素、柠檬酸、β-羟基丁酸、亚油酸、葡萄糖、丙氨酸、油酸和脯氨酸。(3)差异性分析:11种有明显变化的代谢产物差异性分析显示,花生四烯酸、硬脂酸、乳酸、尿素、β-羟基丁酸、油酸在冠心病中均有不同程度提高,以CHD心血瘀阻证中增高最为明显(均P0.01)。其中CHD心血瘀阻证组的乳酸、β-羟基丁酸、亚油酸及葡萄糖值较CHD非心血瘀阻证增高显著(均P0.01);而CHD心血瘀阻证组葡萄糖与健康组比较,差异有统计学意义(P0.01)。柠檬酸在冠心病中呈现不同程度的下降,其中CHD心血瘀阻证组与健康组比较差异有统计学意义(P0.01)。结论 (1)花生四烯酸、硬脂酸、乳酸、尿素、柠檬酸、β-羟基丁酸、油酸、葡萄糖、丙氨酸为CHD心血瘀阻证患者的血浆代谢产物谱。(2)从血浆代谢产物谱分析,CHD心血瘀阻证与脂质代谢、糖代谢相关,同时也与缺氧、剧痛引起的应激相关。

Detection and Analysis on Plasma Metabolomics in Patient with Coronary Heart Disease of Xin-blood Stasis Syndrome Pattern

Objective To research the plasmic metabolites and metabolic pathway of Xin-blood stasis syndrome （XBSS）. Methods Plasma metabolic products in patients of coronary heart disease （CHD） with XBSS or non-XBSS and subjects in the control group were identified by gas chromatographic mass spectrometry （GC-MS） type QP2010,the changes of their main elements in different groups were analyzed by principal components analysis （PCA） and partial least squares （PLS） analysis. Results PCA showed that as compared with that in the control group,in the CHD-XBSS group,contents of lactic acid,β-hydroxy butanoic acid,urea,oleic acid,octadecanoic acid and arachidonic acid were higher and that of citric acid was lower. PLS analysis showed significant difference between the control group and the other two groups,and the latter two groups tend to be of a same category. The occurrence of XBSS was positively correlated with octadecanoic acid,arachidonic acid,urea,lactic acid and β-hydroxy,butanoic acid contents,and negatively correlated with oleic acid,L-proline,glycine,and citric acid contents. According to VIP,the degree of correlation between variables with drug intervention,from high to low,were ranked as arachidonic acid,octadecanoic acid,lactic acid,urea,β-hydroxy butanoic acid,linoleic acid,glucose,alanine,oleic acid and proline. Discrepancy analysis on 11 changeful metabolites showed that the contents of arachidonic acid,octadecanoic acid,lactic acid,urea,β-hydroxy butanoic acid and oleic acid increased in CHD patients,especially in those with XBSS （P0.01）. In CHD patients,contents of lactic acid,β-hydroxy butanoic acid,linoleic acid and glucose in patients of XBSS pattern were higher than in non-XBSS pattern （P0.01）; content of linoleic acid,glucose,alanine and proline decreased in non-XBSS pattern while increased in XBSS pattern. Content of glucose in CHD-XBSS patients was significantly higher than that in the healthy control （P0.01）. Content of citric acid was lower in CHD patients,and showed significant difference between that in CHD-XBSS patients and healthy control （P0.01）. Conclusions The major plasmic metabolites in CHD-XBSS patients are arachidonic acid,octadecanoic acid,lactic acid,urea,citric acid,β-hydroxybutyric acid,oleic acid,glucose,and alanine. Analyzed from plasmic metabolite spectrum view,CHD-XBSS is related with lipid metabolism and glyco-metabolism,also with the stress induced by hypoxia and agonia.