Metabonomics Study Of Ischemic Cerebral Apoplexy Rats Treated With Tongsaimai Pellets

Ischemic Cerebral Apoplexy (ICA) belongs to cerebral vaseular disease in morden medicine. It is one of three death diseases in the world and has high rates of incidence, death, recrudescence and crippledom. For overcoming this disease, treatment based on the theory of Traditional Chinese Medicines (TCMs) was established. However, therapeutic effects of TCMs are integrative results of multiple bioactive components acted on multiple target, as a result there seems to be devoid of effective research method to validate this theory. Currently, metabonomics, a new rapid-developing technology aims at the comprehensive and quantitative analysis of wide arrays of metabolites in biological samples was introduced. Tongsaimai pellets was the study object, and the therapeutic effect of Tongsaimai pellets on ICA was researched from a holistic perspective.In this study, rat model of ICA was established by electric coagulation on middle cerebral artery, and then ICA related bio-fluids’(blood, brain tissue and cerebrospinal fluid) were tested by the method based on the high performance liquid chromatography combined with mass spectrometry and stechiometry analysis technology, meanwhile,we researched three kinds of monoamine neurotransmitters in brain tissue. There are four directions of this article:1Brain metabonomics study of ischemic cerebral apoplexy rats treated with Tongsaimai pelletsRats were divided into4groups, model group, sham-operation group, Tongsaimai pellets group and positive control group. Tongsaimai pellets and positive control group were orally administrated by13.2g·kg-1·d-1of crude drugs and32mg·kg-1·d-1of Nimodipine respectively, model and sham-operation group by equal volume of distilled water for a week. Rat brain tissue of each group were collected on the7th day after administration. Endogenous metabolites of four groups were determined with GC-MS. Then artial least squares discriminant analysis (PLS-DA) was applied to analyze multivariate data, and T-test was used in significant statistical analysis.Scatter plot of rat brain samples in model group was different from3other groups, Compared with sham-operation group rats, alanine, serine, methionine, citric acid, histidine, arachidonic acid, docosahexaenoic acid increased in model group rats, while ribose, glucose-6-phosphate and inosine decreased, so these metabolites were potential metabolic biomarkers. These endogenous metabolites got restoration in Tongsaimai group rats. Arachidonic acid (AA) and docosahexaenoic acid (DHA) were statistically different between Tongsaimai pellets group and model group rats, showing that Tongsaimai pellets could obviously regulate arachidonic acid and docosahexaenoic acid. So changed metabolities can be certainly recovered by Tongsaimai pellets, and the treatment of Tongsaimai pellets can be connected with the regulation of related metabolic pathways. 2Plasma metabonomics study of ischemic cerebral apoplexy rats treated with Tongsaimai pelletsRats were divided and administrated like the first part. Plasma of model and sham-operation group were collected, and plasma of Tongsaimai pellets and positive control group were collected on the1st、3rd and7th day after administration and to be determined with GC-MS.Compared with sham-operation group rats, pyruvic acid, taurine and hydroxyproline obviously increased in model group rats, while lactic acid, glyceric acid, aminomalonic acid, fructose, tryptophan and leucine significantly decreased, so these metabolites were potential metabolic biomarkers. These endogenous metabolites except taurine got restoration in Tongsaimai group rats. The result showed that abnormal metabolite level in plasma can be certainly recovered by Tongsaimai pellets.3Cerebrospinal fluid (CSF) metabonomics study of ischemic cerebral apoplexy rats treated with Tongsaimai pelletsRats were divided and administrated like the first part. CSF of each group were collected on the7th day after administration. Then CSF was determined with GC-MS.Different from brain tissue and serum result, the kind and content of endogenous metabolites in CSF did not change much. Compared with sham-operation group rats, xylose decreased in model group rats but it got restoration in Tongsaimai group rats. Totalion chromatograms of Tongsaimai group was not different from model group on the whole. Glueopyranose was different in Tongsaimai group from model group. So composition of CSF did not change much after modeled or administration.4Effect of Tongsaimai pellets on monoamine neurotransmitter levels in brain tissue of ischemic cerebral apoplexy ratsRats were divided into3groups, model group, sham-operation group and Tongsaimai pellets group. Tongsaimai pellets group was orally administrated by13.2g·kg-1d-1of crude drugs, model and sham-operation group by equal volume of distilled water for3days. Rat brain tissue of each group were collected on the3th day after administration.Compared with sham-operation group rats, NE, DA and5-HT increased in model group rats, showing that these monoamine neurotransmitter contents were higher after modeled, and these monoamine neurotransmitters got restoration in Tongsaimai group rats.5-HT was statistically different between Tongsaimai pellets group and model group rats, showing that Tongsaimai pellets could obviously regulate5-HT.