基于代谢组学的心血瘀阻证动态演变过程研究

Research on Dynamic Evolution of Blockade of Heart Vessel Syndrome Based on Metabonomics

目的研究心血瘀阻证形成过程的代谢产物组学的变化。方法采用高脂饲料与冠状动脉左前降支结扎术结合的方法,制备大鼠心血瘀阻证形成的3个阶段。以血脂增高为血瘀证前期组,动脉粥样硬化形成为亚血瘀证期组,动脉粥样硬化基础之上结扎冠状动脉为心血瘀阻证组,每组8只。另取24只大鼠作为空白对照组,每个阶段8只。应用气相色谱-质谱联用仪(gas chromatography-mass spectrometer,GCMS),对不同组之间代谢产物含量的变化进行主成分分析(principal component analysis,PCA)及偏最小二乘法分析(partial least squares method,PLS)。结果 (1)在鉴定出的32中代谢产物中与心血瘀阻证形成过程密切相关依次是柠檬酸、胆固醇、肌醇、鸟氨酸、脯氨酸、异亮氨酸、硬脂酸、乳酸、尿素、亮氨酸、亚油酸、甘露糖。(2)3个阶段代谢标志物,血瘀证前期:硬脂酸、乳酸(正相关),甘露糖(负相关);亚血瘀证期:鸟氨酸、脯氨酸、肌醇(正相关),异亮氨酸(负相关);心血瘀阻证:亮氨酸、异亮氨酸、柠檬酸(正相关)、乳酸(负相关)。结论血瘀证前期高脂饮食引起体内脂质代谢紊乱,机体启动抗炎;亚血瘀证期持续高脂饮食所导致的尿素循环紊乱、肠道菌群失调、血管形态改变、肝脏功能障碍;心血瘀阻期急性心肌缺血后引起的糖代谢障碍。

Objective To observe the changes of metabolomics in the evolution process of blockade of heart vessel syndrome（BHVS）.Methods The formation of BHVS in three stages were simulated by using high-fat forage and ligating the left anterior descending coronary artery.Increased blood lipid was in the early stage of blood stasis syndrome（BSS）group.Atherosclerosis（AS）was formed in the middle stage of BSS group（sub-BSS）.Coronary artery was ligated on the basis of AS was the 3rd stage of BSS（BHVS group）.There were 8 rats in each group.Totally24 rats was used as the blank control group and each stage had 8 rats.The changes of metabolite contents were analyzed using principal component analysis（PCA）and partial least squares method（PLS）with gas chromatographymass spectrometer（GC-MS）among different groups.Results（1）In the 32 kinds of identified metabolites,citric acid was closest associated with the evolution process of BHVS,followed by cholesterol,inositol,ornithine,proline,isoleucine,octadecanoic acid,lactic acid,urea,leucine,linoleic acid,mannose.（2）Metabolic markers in the threestages：octadecanoic acid,lactic acid（positively correlated）,and mannose（negatively correlated）in the early stage of BSS.Ornithine,proline,inositol（positively correlated）,and isoleucine（negatively correlated）in the middle stage of BSS（sub-BSS）.Leucine,isoleucine,citric acid（positively correlated）,and lactic acid（negatively correlated）in the BHVS stage.Conclusions High fat diet causes disorderedin vivolipid metabolism in pre-stage BSS,and the organism initiates anti-inflammation.Continued high fat diet leads to disordered urea cycle,imbalanced intestinal flora,changed vascular morphology,and liver dysfunction in the sub-BSS stage.Acute myocardial ischemia leads to glucose metabolism disorder in the BHVS stage.