The Research Of Nonalcoholic Fatty Liver Disease Metabonomics By The Treatment Of Danggui Shaoyao Powder Flavored

Objective:Respectively by using GC/MS(gas chromatography mass spectrometry technology) and 1HNMR(nuclear magnetic resonance) on rat liver depression of nonalcoholic fatty liver disease spleen deficiency of blood and urine tests, and then by using principal component analysis(PCA) and partial least squares discriminant analysis(PLS- DA), to look for angelica peony card related metabolites and explore the pathogenesis of nonalcoholic fatty liver disease section, for diagnosis and treatment of nonalcoholic fatty liver disease.Methods:Purchased in different batches SPF healthy male rats, respectively, 24, 32,and GC-MS, nuclear magnetic resonance(NMR) were used respectively to find angelica peony card metabolites. Rats were randomly divided into blank group, model group,treatment group(the first: angelica scattered flavored peony group; the second: Danggui Xiaoyao Powder flavored, Xiao Yao power group), 8, feed grade clean animal room, indoor temperature to keep(22±1) ℃, relative humidity is(46±5)%. Chronic bondage through high sugar、high fat forage、stress hunger disorder and clip tail, establish NAFLD liver spleen deficiency model. After 12 weeks,the treatment group rats will be continuously lavaged by Chinese medicine to medicine dose equivalent conversion method; At the same time, the blank group and model group rats imbedded in capacity of distilled water, such as statistical weight of rats and general behavior during the experiment. All rats after 30 days lavaging, intraperitoneal injection of 10% chloral hydrate anesthesia, abdominal aortic blood, serum, centrifugal conventional preparation in- 80 ℃ low temperature refrigerator.Set aside. Remove liver tissue, and is divided into two parts, one with physiological saline rinse put in formaldehyde fixed standby, and the other one will be remained a placed-80 ℃ refrigerator. Through liver homogenate pathological index verification model, after the preparation of samples, named after the metabolites of using GC-MS, 1 h NMR technology dealing with blood and urine samples, the RAW format Xcalibur into net CDF format, map after XCMS software pretreatment, to face the normalized data into Excel,and USES the metabonomics software SIMCA P13.0 by load diagram metabolites in VIP value to find the differences. Using SPSS17.0 data processing, the use of groupsindependent samples t test, single factor analysis in the group, with p < 0.05 showed significant difference.Results :(1)Significant changes in behavior can be seen during the experimental period, abnormal pathological index of rats in model group, indicating that nonalcoholic fatty hepatitis liver stagnation and spleen deficiency syndrome model validation is good, as an animal model of reference; supplemented Danggui Shaoyao Power, Xiaoyao Power can regulation and biochemical indexes, effectively treat the liver stagnation and spleen deficiency syndrome of Nonalcoholic fatty liver disease.(2) GC/MS results: Concluded three different metabolites, single palmitic acid glyceride, oxalic acid, two imine carbide.Compared with the blank group, model group serum single palmitic acid glyceride, lactic acid, oxalic acid, two imine carbide, 3- hydroxybutyrate, glycine, furan e indican,galactose and glucose, methylamine,18 kinds of metabolites are changing. Blood classification, main metabolites can be divided into: sugar, galactose and glucose, arlo sugar; Amino acids: glycine; Lipid classes: single palmitic acid glyceride; Fatty acid,palmitic acid; Organic acid, oxalic acid, glycolic acid, lactic acid, 3- hydroxybutyrate;Other categories: two imine carbide, methylamine, etc.(3)1HNMR results:(1)Blood: in the model group 23 metabolites levels will be significantly abnormal, and they can be changed significantly tending to be the normal level after treatment: amino acids, glycine, lysine ammonia acid, glutamic acid, arginine, histidine, threonine, dimethyl glycine; fatty acid:citric acid, 3-hydroxy butyric acid, lipid creatine; lipid glycerol, triglyceride; organic alcohols: scyllo inositol, myo inositol; sugar: glycogen, alpha glucose; organic base class:Betaine, acetyl glycoproteins, methylamine, triple oxide methylamine, choline; other classes: formic acid, and acetic acid.(2)Urine: compared with the blank group, the model group found that there are differences in the level of 27 metabolites, and the two prescriptions can effectively regulate these metabolites and tend to be normal, and there is statistical significance. It is respectively: amino acids, glycine, lysine ammonia acid,glutamic acid, proline, arginine, alanine, threonine, isoleucintryptophan, glutamine;organic acid: asparagine, amino adipic acid, citric acid, lactic acid, pyruvic acid, acetic acid,3- hydroxy butyric acid, acetic acid; sugar: alpha glucose; organic alkali: Betaine, lipid glycerine and glycoprotein lipid, phenyl acetate; other classes: acetone, methylamine,trimethylamine oxide.Conclusion:(1) Model validation is good, and two groups of model group rats, by the treatment of Danggui Shaoyao Powder flavored after 30 days, all can show the weight gain, increased appetite, grooming times increase, such as the colour rotary normal behavior performance.(2) GC/MS, according to the results of nonalcoholic fatty liver disease, liver spleen deficiency and lipid metabolism, energy metabolism, inflammation,and immune related, and angelica peony flavored to regulate these, by for nonalcoholic treatment effect. The related markers by 1HNMR, is concentrated on expression in lipid,energy metabolism, amino acid metabolism, IR, inflammatory reaction.( 3) Danggui Shaoyao powder flavored and Xiaoyao powder can reduce lipid peroxidation and reduce the degree of IR through regulating lipid metabolism, energy metabolism. Among them,the Danggui Shaoyao powder flavored is mainly to improve the energy metabolism in model group, improve the utilization rate of glucose to reduce IR; Xiaoyao Powder is primarily to regulation of amino acids, reduce oxidative stress and lipid oxidation degree to reduce the IR.(4)Two experimental results, it can be for the study of NAFLD in IR provides the experimental basis for the pathogenesis and clinical treatment.