Study The Mechanisms Of Couplet Medicines Gualou-Xiebai On Chronic Myocardial Ischemia Based On Metabonomics

Objective:To study the therapeutic effects of Gualou-Xiebai on chronic myocardial ischemia rats induced by ISO and the effects of Trichosanthes kirilowii Maxim and Allii macrostemi bulbus alone on chronic myocardial ischemia rats by using ~1H NMR metabolomics,and to explore the differential metabolites and related metabolic pathways,and to explore the effect of Gualou-Xiebai couplet medicines on chronic myocardial ischemia.Mechanism.Methods:Thirty SD male rats were randomly divided into five groups:Gualou group,Xiebai group,Gualou-Xiebai group,model group and normal group,with 6 rats in each group.Chronic myocardial ischemia model was induced by long-term administration of isoproterenol ISO at low doses in rats.The chronic myocardial ischemia model was established by intraperitoneal injection of ISO 0.04 mg/kg for 6 days in Gualou group,Xiebai group,Gualou-Xiebai group and model group respectively,and 0.04 mg/kg of normal saline was injected intraperitoneally.The changes of electrocardiogram(ECG)and ST segment voltage of rats in each group were recorded after the model was established.After 14 days of continuous gastric administration,blood and heart tissues of rats were taken for pathological electron microscopic observation.Metabolites in serum and heart of rats were detected by ~1H NMR technology.Differential metabolites were screened out.Related pathways were found according to different metabolites.The change of substance content responds to the regulation of metabolic pathway.Results:(1)After the establishment of the model,we can see that the fluctuation of ECG in the model group and each administration group is very obvious,and the voltage of ST segment is obviously increased,suggesting that the model is successful.(2)After HE staining,the rat hearts were observed under optical microscope.The results showed that the cell structure of normal group was clear,the muscle fibers arranged neatly,the cell structure of model group was blurred,disordered,a large number of inflammatory cells infiltrated,and a large number of vacuoles were observed in the cells;the cells arranged closely,and obvious edema was observed.Cell edema and inflammation were improved in each group after administration,but cell structure was clearer and myocardial fibers were arranged neatly than other groups,which was very close to the normal group,and was quite different from the model group.It showed that the effect of Gualou-Xiebai was better than that of single use.(3)Seven potential biomarkers were found in cardiac tissues based on ~1H NMR spectra,literature and corresponding library:sarcosine,inosine,inosine,glycerol,glucose,adenosine diphosphate and adenosine triphosphate.The metabolic pathways of chronic myocardial ischemia rats were analyzed.The main metabolic pathways were purine metabolism and galactose metabolism.However,the effect of Gualou-Xiebai group on myocardial ischemia was very significant(P<0.01)by decreasing inosine content and increasing adenosine triphosphate,adenosine diphosphate,adenosine phosphate and hypoxanthine nucleotide content,so as to regulate the purine metabolic pathway.(4)Data processing and analysis of rat serum revealed eight potential biomarkers:valine,tyrosine,threonine,serine,phenylalanine,isoleucine,glutamine and 3-hydroxybutyric acid.The main metabolic pathways of chronicmyocardialischemiawere:aminoacyltRNA biosynthesis,phenylalanine,tyrosineandtryptophanbiosynthesis,phenylalanine metabolism,valine,leucine and isoleucine biosynthesis,glycine,serine and threonine metabolism.The content of ketoisohexanoic acid,3-hydroxybutyric acid,glutamic acid,isoleucine,phenylalanine,succinic acid,taurine and valine was up-regulated in the Gualou-Xiebai group by regulating the biosynthesis of valine,leucine and isoleucine,the degradation of valine,leucine and isoleucine,the metabolism of alanine,aspartate and glutamic acid,phenylalanine,tyrosine and tryptophan.Biosynthesis,D-glutamine and D-glutamine metabolic pathways were effective in the treatment of myocardial ischemia(P<0.05).Conclusion:Gualou-Xiebai has therapeutic effect on chronic myocardial ischemia rats and its single drug.Compared with the single medicine,the treatment effect of Gualou-xiebai is more obvious.Many metabolic pathways have been changed in rats with myocardial ischemia treated by trichosanthin.By regulating purine metabolism,valine,leucine and isoleucine biosynthesis,valine,leucine and isoleucine degradation,alanine,aspartate and glutamic acid metabolism,phenylalanine,tyrosine and tryptophan biosynthesis,D-glutamine and D-glutamine metabolism,etc.The pathway achieves the goal of treating chronic myocardial ischemia.