大鼠心血瘀阻证心肌组织代谢组学的研究

目的寻找心血瘀阻证大鼠心肌组织代谢产物谱,以及养心通脉方干预后表型的变化,分析大鼠心血瘀阻证心肌组织的代谢途径。方法采用结扎大鼠冠状动脉左前降支复制急性心肌梗死(AMI)心血瘀阻证模型,同时采用养心通脉方干预模型,运用GC-MS检测模型组、养心通脉方组、假手术组和空白对照组4组大鼠心肌组织代谢产物,每组8只,通过对不同组之间代谢产物含量的变化进行主成分分析、偏最小二乘法分析。结果主成分分析:四组样本主成分得分分析,模型组、养心通脉方组与正常组、假手术组分聚于椭圆形散点图的3个区域。因子载荷图显示模型组甘氨酸、延胡索酸、苹果酸、谷氨酸、葡萄糖、磷酸、吡喃半乳糖、赖氨酸发生了改变。偏最小二乘法分析:PLS回归模型显示,模型组与养心通脉方组存在明显的线性关系,证明模型建立是合理的。药物干预与甘氨酸、苹果酸、谷氨酸、葡萄糖高度正相关,与尿素、琥珀酸高度相关,与赖氨酸负相关。按照VIP值依次排列各变量与药物干预的密切度依次是苹果酸、谷氨酸、甘氨酸、葡萄糖、延胡索酸、尿素、半乳糖、酪氨酸、乳酸、丙氨酸。差异性分析:明显变化的代谢产物差异性分析表明,甘氨酸等10种代谢产物在模型组中显著下降,与正常组比较差异有统计学意义(P<0.01)。药物干预后甘氨酸、延胡索酸、苹果酸、谷氨酸、半乳糖、葡萄糖、酪氨酸、尿素、乳酸、丙氨酸等含量增加与模型组比较差异有统计学意义(P<0.01)。其中苹果酸、谷氨酸、酪氨酸、尿素上升幅度较小,与正常组比较差异仍有统计学意义。赖氨酸经药物干预后均数显示有轻度下降,但与模型组比较差异无统计学意义。大鼠AMI心血瘀阻证与心肌组织代谢产物苹果酸、谷氨酸、甘氨酸、葡萄糖、延胡索酸、尿素、吡喃半乳糖、乳酸、丙氨酸、酪氨酸等含量的改变密切相关。结论心肌组织代谢产物谱显示AMI心血瘀阻证代谢途径与缺氧后的糖代谢紊乱密切相关,养心通脉方可以有效干预这一过程。

Metabolomics Study of the Myocardial Tissue of Rats of Cardiac Blood Stasis Syndrome

Objective To find out the metabolite profile of rats’ myocardial tissue of cardiac blood stasis syndrome(CBSS),and to analyze the metabolic pathway of CBSS rats’ myocardial tissue by observing the changes of phynotypes intervened by Yangxin Tongmai Recipe(YTR).Methods Acute myocardial infarction(AMI) rat model of CBSS was prepared by ligating the left anterior descending coronary artery.Meanwhile,the model was interfered with YTR.The metabolites of rats’ myocardial tissue were detected in the model group,the YTR group,the sham-operation group,and the blank control group using GC-MS(8 rats in each group).Changes of metabolite contents were analyzed among different groups using principal component analysis(PCA) and least-square analysis.Results As for PCA:The results of PCA showed that principal component integral(PCI) of the four groups was mainly distributed in the three regions of oval scatterplot.The factor loading gram showed that contents of glycine,fumaric acid,malic acid,glutamic acid,glucose,phosphoric acid,galactopyranose,lysine were changed in the model group.Analysis of partial least square method:PLS regression model showed that obvious linear correlation existed between the model group and the YTR group,which proved the model was reasonably established.The drug intervention was highly positively correlated with glycine,malic acid,glutamic acid,glucose,highly correlated with urea and butanedioic acid,but negatively correlated with lysine.According to VIP value,each variable was closely correlated with the drug intervention in sequence as malic acid,glutamic acid,glycine,glucose,fumaric acid,urea,galactose,tyrosine,lactic acid,and alanine.Results of variability analysis:Obvious changed variability analysis of metabolite difference showed that 10 metabolites such as glycine,etc.obviously decreased in the model group,showing significant difference when compared with the normal group(P<0.01).Compared with the model group,contents of glycine,fumaric acid,malic acid,glutamic acid,glucose,tyrosine,urea,lactic acid,and alanine,etc.obviously increased after drug intervention(P<0.01).Of them,the increment of malic acid,glumatic acid,tyrosine,and urea was less,showing significant difference when compared with that of the normal group.The mean of lysine was slightly lowered after drug intervention,but with insignificant difference when compared with that of the model group.AMI rats of CBSS was closely correlated with myocardial metabolites such as malic acid,glutamic acid,glycine,glucose,fumaric acid,urea,galactopyranose,lactic acid,alanine,and tyrosine,etc.Conclusions The metabolite profile of rats’ myocardial tissue showed AMI rat model of CBSS was closely correlated with post-hypoxia glucose metabolism disorder.YTR could effectively intervene this process.