Based Metabolomics Spleen Blood Deficiency Rat Model Of Metabolic Syndrome

Purpose：Metabonomicsisascienceresearchaboutorganismsofwhosemetabolitetypes, quantity and its regular change after disturbance.Focus on exploring thebiological of whole, organ or tissue of the endogenous metabolites in metabolicpathways, and the intrinsic and extrinsic factors influencing metabolism ofwhole trajectory of change, in order to reflect a certain pathophysiologicalprocesses that occur in a series of biological events.With the key tocharacteristics of its holism and system, which is associated to the methodologyprinciple in traditional Chinese medicine is unity, i.e. both of two holism andsystem are recognized as the starting point for cognition, so can metabonomicstheory and technical research of syndrome essence in TCM as the starting pointand the starting point.The experimental research, through establishment ofspleen yang deficiency rat model of metabolic syndrome,combing treatment basedon syndrome differentiation in TCM and metabonomics research method, on themodel of blood samples for analysis, trying to find characteristic of lesionsmetabolites of spleen yang deficiency of metabolic syndrome, observation ofmetabolic changes and relating physiological with pathological, exploringspleen yang deficiency of metabolic syndrome pathological molecular mechanisms,so as to further reveal the essence of spleen yang deficiency of metabolicsyndrome.Material and method：1. Establishment of animal disease model40healthy male SD rats, weighing (180±10)g, before the experiment adaptivefeeding 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=30).In the blank control group, they daily fed ordinary standard rat dietand drank water which were free.In the model group,each rat daily fed cabbage15～20g, swam10min; doubleday gave high-fat and high-sugar diet.There was adaily to drench100%ice senna water1ml/100g;the whole time of12weeks.2. The research of metabonomics The research of metabonomics selected randomly six rats which modeledsuccessfully with spleen yang deficiency of metabolic syndrome and six rats inthe blank control group.The used of the USA Waters company Ultra PerformanceLiquid Chromatography and Quadrupole-Time-of-Flight Mass Spectrometry(UPLC-Q-TOF-MS), the obtained original mass spectrometry data into MarkerLynxsoftware package,for peak identification, peak alignment and baselinecorrection of pretreatment, the final output matrix, i.e. accurate karyoplasmicratio and retention time of variable composition, sample name and peakstrength/area.The processing of a data matrix into Simca-P software (version11.5) and the metabolic finger graph used PCA,PLS-DA and OPLS-DA, to find adifference characteristic metabolites, thereby identification ofbiomarkers,tracing its sources and metabolic pathways.Results：1. Establishment of animal disease modelThe rats in the model group compared with the blank control group, aftertwo weeks, manifested as reduced activity, fatigue, gained weight, loose stool,perianal region with dirt and spirit of cachexia. At the same time,body hairbrightness gradually subsided and swimming endurance fell daily.From thebeginning of the eighth week, body weight of the rats in the model group comparedwith the blank control group increased obviously.And there was significantdifference between two groups (P<0.05). After12weeks,triglyceride, fastingblood glucose,insulin and HOMA-IR of the rats in the model group compared withthe blank control group, increased significantly.And there was significantdifferences between two groups (P<0.05). After12weeks, high densitylipoprotein of the rats in the model group was lower than that. And there wassignificant difference between two groups (P<0.05).At the end of modeling,therats in the model group were consistent with the experimental research whichwas designed to establish evaluation standard of spleen yang deficiency andmetabolic syndrome.2. The research of metabonomicsBy PLS-DA and OPLS-DA, blood metabolites of the rats in the model group andthe blank control group were distinguished well.And two groups showed no crossand overlap obviously.3. Potential biomarkersThe research found potential biomarkers which associated with spleen yangdeficiency rat model of metabolic syndrome.They mainly wereN-acetyl-D-glucosamine, homocysteine,lysophosphatidylcholine, diglyceride and so on.The material collection was characteristics of metabonomics which wasspleen yang deficiency rat model of metabolic syndrome.Conclusion：1．Imbalance of eating, overstrain which harmed spleen qi and bitter and coldin property causing diarrhea which harmed spleen yang reached the purpose offirst loss of spleen qi and then the damage of spleen yang.After that,the ratscontinuously were fed high-fat and high-sugar diet for12weeks.At last,theywere successfully established spleen yang deficiency rat model of metabolicsyndrome.2. By metabonomics,the experimental research observed blood metabolites graphsof spleen yang deficiency rat model of metabolic syndrome.The graphs weredifferent from distributional graphs of the normal rats.3. The research found potential biomarkers which associated with spleen yangdeficiency rat model of metabolic syndrome.They mainly wereN-acetyl-D-glucosamine,homocysteine,lysophosphatidylcholine, diglycerideand so on.The research revealed that the spleen yang deficiency of metabolicsyndrome related to the disorder of carbohydrate, lipid and proteinmetabolic.And these internal small molecule compounds may be the material baseof syndrome in traditional Chinese medicine of metabolic syndrome.4. Metabonomics which researches into the mechanism of diseases and syndromeessence in traditional Chinese medicine in depth is the important method.