Studies On The Molecular Mechanism Ofrhubarbcoptis Chinensis Decoction Againstcerebral Ischemia Based On Metabolomics

At present,cerebral apoplexy,as the second most fatal disease in the world,poses a great threat to national health due to its characteristics of high regional morbidity,high disability rate after onset and high recurrence rate after treatment.Treatise on febrile disease rhubarbcoptis chinensis decoction has a good protective effect on brain in modern pharmacology research,while rhubarbcoptis chinensis decoction have outstanding performance in single drug use and high frequency in clinical application.In addition,modern Chinese medicine studies on the pathogenesis of "toxic damage to cerebral collaterals" are highly respected.This theory demonstrates that heat toxicity is one of the key causes of stroke,among which clearing heat,purging fire and detoxifying drugs can significantly alleviate the disease.Because rhubarbcoptis chinensis decoction has a good protective effect on brain in clinic,but the molecular mechanism of rhubarbcoptis chinensis decoction is not clear.This paper will with rhubarb soup as the research object,rhubarbcoptis chinensis decoction game focal cerebral ischemia in mice to fill the stomach to medicine,combined with nerve factor score,effectiveness indexes such as cerebral infarction area and brain water content,and then introduce the metabonomics technology research of the mechanism of cerebral ischemia,through to the mice serum and brain tissue under different condition detection,pattern recognition,data inquiry rhubarbcoptis chinensis decoction against cerebral ischemia mechanism of rhubarbcoptis chinensis decoction,mining related endogenous metabolites and metabolic pathways,discuss the related molecular mechanism.It provides a new way for Chinese medicine to prevent cerebral ischemiaMain research methods and results are as follows:(1)The mouse model of focal cerebral ischemia was established by using the method of thread plug,and the mice were given the drug by intragastric intragastric administration for seven days.The life state of mice was observed and the degree of nerve function was distinguished according to Longa neurological function score.TTC staining method and Image recognition by Image J-Pro PLUS 6.0 software were adopted to measure the cerebral infarction area.,the degree of cerebral edema was combined with the method of determination of cerebral water content,and the anti-cerebral ischemia effect of rhubarbcoptis chinensis decoction was evaluated.Concentration is 2.4 g/ml of rhubarbcoptis chinensis decoction treatment effect is not significant,and give the dose of 1.2 g/ml and 0.6 g/ml show the obvious good prevention effect,expressed as body recovery of neurological function,cerebral infarction area,and compared with model group obviously alleviate cerebral water content,and presented the effects of rhubarbcoptis chinensis decoction concentration-response relationship is nonlinear relationship.(2)Effective doses ofrhubarbcoptis chinensis decoction were 1.2 g/ml and 0.6 g/ml according to the pharmacodynamic indexes of different doses.The serum and brain tissues of mice in different states were analyzed and determined by gc-ms technology,and endogenous metabolites were identified by combining with NIST 08 database and HMDB database.Using MetaboAnalyst 4.0 software by pattern recognition in serum and brain tissue data analysis of principal component analysis,partial least squares methods of exploratory analysis,multivariate ROC curve analysis,control group and model group sample points are completely separate state,confirmed that the focal cerebral ischemia in mice building successful replication,and focal cerebral ischemia model have interference effects on the body's metabolism in mice.The analysis of data samples of mice in different states suggested that rhubarbcoptis chinensis decoction had the function of relieving nerve function injury in mice,and suggested that the body of mice had the tendency of moderating to normal signs.In model group and rhubarbcoptis chinensis decoction group of mice serum and brain tissue fluid is the relevance of the endogenous substances,were found control,rhubarbcoptis chinensis decoction in low dose group compared with model group,the change of the relative content of endogenous substance has the similarity,namely compared with control group,model group is greater than the difference of therhubarbcoptis chinensis decoction group.According to VIP>1 combined with non-parametric test P<0.05,12 and 7 different metabolites in serum and brain tissue were screened.The Impact>0.10 pathway was selected,and differential metabolites analysis of serum and brain tissue revealed 9 and 2 metabolic pathways.Pathway Analysis database was used for topological Analysis of differential metabolites,and it was speculated that the intervention effect of rhubarbcoptis chinensis decoction group might be related to the metabolism of glutamine and glutamate,arachidonic acid and carbohydrate pathways.A modular structure was constructed by Cytoscape to analyze the correlation of metabolic endogenous.The correlation analysis and modular analysis of serum and brain tissue differential metabolites found that there were close relations between 3 and 4 modules,and the intervention effect of rhubarbcoptis chinensis decoction group might be related to the differences of metabolites in 3 and 4 modules of serum and brain tissue.