The Stagnation Of Liver-Qi Syndrome Experiment And Clinical Study Based On Metabonomics

Objective:The stagnation of Liver-Qi syndrome is a very far-ranging and pervasive syndrome, and also a familiar TCM (Traditional Chinese Medicine) internal medicine syndrome. According to sample statistic of TCM internal medicine history cases, around 21% patient get the stagnation of Liver-Qi syndrome. Recent years, entitative biological study to the stagnation of Liver-Qi syndrome is basically study of single system, which covers aspects of Blood Biochemical, autonomic nerve function, Trace Element Level, Blood rheology study, molecular biology, sex hormone, Damage of Organs, and Parametric Testing, etc., but few study is based on multi-system aspect. There is no illustrated biological foundation for the stagnation of Liver-Qi syndrome. Metabonomics is nowdays hot topic in biology and iatrology, which is used as quantitative analysis for almost all metabolites of all kinds of familiar organism, and describe the parameter dynamic change of metabolites, in order to find the relationship of metabolites change and pathophysiology, so that to study the macrocosm and the all, through studying different effect factor of the whole-body system, system and Apparatus etabolism approach and Endogenous etabolism matter. It is a representation of Tradicitional Chinese Medicine Holism, that apply metabonomics to The stagnation of Liver-Qi syndrome. In this study, on the one hand we use emotion stimulating and pinching the tail method to make rat model of stagnation of Liver-Qi syndrome, and intervent by Bupleurum smoothing liver medicine, observe the modification before and after take the medicin. On the other hand, the cilinic aspect, we choose patients with Ganqi stagnation, observe the change of emiction and blood metabolite, from the point of metabonomics, to holistic expound the biological fundation of stagnation of Liver-Qi syndrome, in order to analysis the Phenotype Characteristics and possible biological fundation of the stagnation of Liver-Qi sydrome, from a systemic and holistic Small molecule metabonomics level, and also provide relavant theory and experiment support for further prevention for the stagnation of Liver-Qi syndrome.Method:30 healthy SD rat, body weight 220g, divide to three groups by chance:Group A:Normal 10; Group B:model group 10; Group C: medicine group 10. Except for group A, the other two groups was made into model of the stagnation of Liver-Qi syndrome. Group C was provided Bupleurum smoothing liver medicine,0.5g/mL, twice per day. All groups is to collect emiction and blood samples the day before modelling, after modeling, and after take the medicine, go through NMR Spectrometer examination, and use metabonomics method, after observe group A, B and C, the change of emiction and blood Small-molecule metabolites, and meanwhile observe emiction and blood metabonomics study of 30 clinical patients with Ganqi stagnation and 30 healthy people, and study the biological fundation and pathophysiology, from the point of metabonomics.Result:一,Animal experiment study(1) Comparing with the normal group, the model rat emiction Hippuric acid content 3.97.7.55,7.85, a-Ketoglutarate 3.01,2.45, citrate2.57,4.10,2.88, isocitrate 3.99 2.51 2.45and aconitate 3.77 6.96 3.96 the curve relative integral acreage obviously decrease, and decrease in emiction, Creatinine 3.05,4.07, acetone 2.23, acetic acid 1.93, reatine 3.93,3.03, nicotinate 4.41, 1.41 and acid5-hydroxyindole-3-acetate,5-HIAA 3.03,2.91,6.63, 7.15 the curve relative integral acreage obviously increase, and increase in emiction.(2) After intervene by Bupleurum smoothing liver medicine, the Metabolic phenotype of group C was reatine 3.93,3.03, acetone 2.23 Creatinine 3.05 the content was obviously decreased, N-TMAO 4.07, TMAO 3.27, aurine 3.43content was obviousaly increased.(3) From normal group and model group rat blood comparison PCA analysis, the metabolic phenotype of model group is Lactic acid (1.31,1.32,4.11),N-O-GlcNAc(Nac2.02,2.05,2.06,2.09), Choline, saturated fatty acid, B-dextrose(3.47,3.91) increase, unsaturated fatty acid, HDL (0.83), Glu, Gln(2.43) Val (1.03,0.98), VLDL (1.29,0.87), LDL, (1.25,0.85) Lipoid (1.27), content was decreased。The symbol of metabolite may be lactic acid, Choline, NAC, saturated fatty acid, blood sugar, unsaturated fatty acid, HDL。(4) The rats from Liver Qi Stagnation Syndrome group were treated by Chaihu Shugansan. After the PCA analysis on the blood-lH-NMR spectral peak integral values, the obtained score plot factor loading diagram (loadingplot), and PC integral values distributed on the oval-shaped scatter (95% confidence region) in four regions, but the two groups can not be distinguished, which indicated that the Phenotypic differences in metabolism of blood compounds of model group and treatment group could not be found.二,clinical study(1) It can be see from the results of comparative PCA analysis on the urine of normal group and Liver Qi Stagnation patients'group that, Liver Qi Stagnation group's urine metabolic phenotype was: creatinine 3.06,4.06,4.1 rose, citrate 2.57,2.71,2.66, Phe (phenylalanine) 3.12, tryptophan (tryptophan) 3.32, acetyl citric acid (acetyl citric acid) 1.28,1.26, tyramine (tyramine) 2.71,2.53, aconitic acid (aconitic acid) 3.17 3.45,3.77 and other compounds decreased.(2) It can be seen from the statistical analysis results of the normal group and Liver Qi Stagnation patients'group that, the PCA analysis on two groups of blood plasma metabonomics showed that there is no distinction of PC between the normal group and the group of liver qi stagnation, which indicated that Liver Qi Stagnation Syndrome patients' endogenous metabolism of plasma spectra did not change significantly.ConclusionIn Liver Qi Stagnation Syndrome, the performance of amino acid metabolism was apparent; citric acid content in urine decreased; creatinine increased; blood lactate concentration increased; glutamic acid, Gln and Screenshot acid declined. The amino acid metabolism differences may be related to liver qi stagnation. Liver Qi Stagnation syndrome is related to energy metabolism, and its compounds include creatinine (ereatinine), lactate (laetate), citric acid (eitrate) and alanine (alanine); Liver Qi Stagnation syndrome is related to neural System Regulation:it was found in the sample patients with the Liver Qi Stagnation Syndrome that a series of metabolites of phenylalanine and tyramine content related with tyrosine metabolism decreased. It was found that Tryptophan may be neural basis for regulating the biological material for Liver Qi Stagnation Syndrome; Liver Qi Stagnation Syndrome group also may have weakened immune function:the metabolites is L-tryptophan metabolism; tryptophan is the Synthesis of 5-HT precursor substance; decrease of tryptophan metabolism resulted in the reduction of 5-HT synthesis; the activation of the sympathetic nervous system regulated the immune system mainly based on suppression, which led to the release of glucocorticoids from the adrenal cortex and the inhibition of immune cells.From the findings above, the changes of biological foundation of Liver Qi Stagnation Syndrome metabolite were found; the mechanism changes of Liver Qi Stagnation organism' s physiological and pathological basis was obtained. Therefore, the study was a successful exploration of the biological features of Liver Qi Stagnation from an overall perspective of metabolomics.