Metabonomics Of Metabolic Syndrome In Rats With Phlegm And Blood Stasis Syndrome

Purpose: The experimental research,through establishment of the rat model of metabolic syndrome of phlegm and blood stasis syndrome.The blood samples of the model were analyzed based on the technology of the metabolic group to try to find characteristic of lesions metabolites in the rat model of metabolic syndrome of phlegm and blood stasis syndrome and observe the metabolic changes and relating physiological with pathological, explore rat model of metabolic syndrome of phlegm and blood stasis syndrome pathological molecular mechanisms, so as to further reveal the essence of the rat model of metabolic syndrome of phlegm and blood stasis syndrome.Material and method: 1.Establishment of animal disease model 38 healthy male Wistar rats of SPF grade, weighing(180±10)g, before the experiment adaptive feeding for one week, free food, drinking water which were no adverse reaction, were randomly divided into the blank control group(n=10)and the model group(n=28). In the blank control group, they daily fed ordinary standard rat diet and drank water which were free.In the model group, each rat daily fed high fat and high glucose food.After 16 weeks, the model group was injected with small dose of STZ to Establishment Rat model of metabolic syndrome of phlegm and blood stasis syndrome. The whole modeling time lasted for 20 weeks. 2.The research of metabonomics The research of metabonomics selected randomly eight rats which modeled successfully with the syndrome of phlegm and blood stasis syndrome and eight rats in the blank control group. GC/TOFMS analysis was performed using an Agilent 7890 gas chromatograph system coupled with a Pegasus HT time-of-flight mass spectrometer. Chroma TOF4.3X software of LECO Corporation and LECO-Fiehn Rtx5 database were used for raw peaks exacting, the data baselines filtering and calibration of the baseline, peak alignment, deconvolution analysis, peak identification and integration of the peak area. And normalized peak area were fed to SIMCA software package(V14, Umetrics AB, Umea, Sweden) for principal component analysis(PCA), partial least squares discriminant analysis(PLS-DA) and orthogonal projections to latent structures-discriminant analysis(OPLS), to find out the characteristic metabolites with different characteristics and then identify potential biomarkers.Results: 1.Establishment of animal disease model The rats in the model group compared with the blank control group, after four weeks weight increased significantly. after six weeks Shit smelled heavy, spirit of cachexia. At the same time,body hair brightness gradually subsided and tongue ecchymosis. Building after 8 weeks,the content of blood lipid in model group was significantly changed compared with that in control group.And there was significant differences between two groups(P<0.05). High blood glucose, high blood insulin and insulin resistance were found in the model group after the model was finished. At the end of modeling,the rats in the model group were consistent with the experimental research which was designed to establish evaluation standard of metabolic syndrome of phlegm and blood stasis syndrome. 2.The research of metabonomics Through the metabonomics technology research, it can be very good to distinguishthe control group and the model group rats blood metabolites, two groups have obvious differences. 3.Potential biomarkers The research found potential biomarkers which associated with rat model of metabolic syndrome of phlegm and blood stasis syndrome.They mainly were N- acetyl-Dglucoseamine,3- hydroxy acid, homocysteine, valine, diacylglycerol such as more than 20 kinds of metabolites. The main markers together constitute the metabolic syndrome of phlegm and blood stasis syndrome model of metabolic syndrome. These substances are the result of the body protein, carbohydrate, lipid and other metabolic disorders.Conclusion: 1. The model of metabolic syndrome of phlegm and blood stasis syndrome can be successfully reproduced by long-term eating high-fat diet,and the method of STZ injection combined with multiple small doses of injection. 2. By metabonomics, the experimental research observed blood metabolites graphs of phlegm and blood stasis Rat model of metabolic syndrome.Finded out the potential biomarkers associated with the syndrome of phlegm and blood stasis syndrome in the rat model. 3.The research found potential biomarkers which associated with rat model of metabolic syndrome of phlegm and blood stasis syndrome.They mainly were N- acetyl-Dglucoseamine,3- hydroxy acid, homocysteine, valine, diacylglycerol such as more than 20 kinds of metabolites. The main markers together constitute the metabolic syndrome of phlegm and blood stasis syndrome model of metabolic syndrome. These substances are the result of the body protein, carbohydrate, lipid and other metabolic disorders. 4.Metabonomics technology can better reveal the essential connotation of disease of TCM syndrome models, The application of metabonomics technology has a profound significance for the modernization of traditional Chinese medicine.