Metabonomics Studies On Blood Deficiency Mice Model Intervened With Angelica Sinensis And Its Different Processed Products

Traditional Chinese(veterinary) medicine(TC(V)M) lays stress on the integrity and efficacy has the characteristics of multi-component, multi-pathway, and multi-target and so on, so we must study the relationship between hemopenia and the enriching blood effect of Angelica sinensis(AS). In this study, water decoction from unprocessed Angelica sinensis(UAS) and its different processed products were selected as the research objects; the blood deficiency mice model was built with acetyl phenylhydrazine(APH) and cyclophosphamide(CTX); gross appearance combining with the routine blood test were used to determine blood deficiency; the physiological and biochemical index of mice were tested, the thymic tissue section was observed, and the blood deficiency different metabolites were selected with metabonomics technology based on Gas chromatography mass spectrometry(GC-MS) and Liquid chromatography- quadrupole-time of flight-mass spectrometry(LC-Q/TOF-MS). Then the reliability of different metabolites selection method was tested and verified with correlation analysis and heatmaps analysis. The obtained different metabolites were inputted into the Met PA metabolic pathway analysis software to study the enriching blood mechanism of UAS and its different processed products; finally the data integration analysis was made to provide a basis for the diagnosis and treatment of blood deficiency. The main results were as follows:1. UAS and its different processed products could improve the clinical symptoms, the decline of blood routine index, the decline of body quality and viscera index, and the decline of ATP enzyme, 6-glucose phosphate dehydrogenase(G6PD), glutathione peroxidase(GPX) and glutathione reductase(GR) activity, and the high methemoglobin content, and the compensatory increase of erythropoietin(EPO) content in the blood in the blood deficiency mice in different degrees. AS parched with alcohol(AAS) had the best enriching blood effect, and followed by AS parched with oil(OAS), AS parched with soil(SAS), UAS and charred AS(CAS).2. The integrated metabolomics research on plasma, liver tissue, spleen tissue and urine of the blood deficiency mice using GC-MS showed that the mechanism of enriching blood of UAS and its different processed products was to restore the Alanine, aspartate and glutamate metabolism in the blood deficiency mice, and to restore D-Glutamine and D-glutamate metabolism, Glycine, serine and threonine metabolism, Arginine and proline metabolism, Glycolysis or Gluconeogenesis, Pyruvate metabolism, and Citrate cycle(TCA cycle), and to restore Cysteine and methionine metabolism, Valine, leucine and isoleucine metabolism, Fructose and mannose metabolism, Amino sugar and nucleotide sugar metabolism, Glycerolipid metabolism, and its related Inositol phosphate metabolism, and finally the enriching blood effect was achieved. 3. The integrated metabolomics research on liver tissue, spleen tissue and urine of the blood deficiency mice using LC-Q/TOF-MS showed that the mechanism of enriching blood of UAS and its different processed products was to restore Glutathione metabolism, Pentose phosphate pathway, Glycine, serine and threonine metabolism and Amino sugar and nucleotide sugar metabolism in the blood deficiency mice, and then to restore Cysteine and methionine metabolism, Taurine and hypotaurine metabolism, Arginine and proline metabolism, D-Glutamine and D-glutamate metabolism, Alanine, aspartate and glutamate metabolism, Histidine metabolism, which was related to Glutathione metabolism; and to restore Glycolysis or Gluconeogenesis, which was related to Pentose phosphate pathway; and to restore Glycerophospholipid metabolism, Sphingolipid metabolism, TCA cycle, Inositol phosphate metabolism, which was related to Glycine, serine and threonine metabolism; and to restore Fructose and mannose metabolism, which was related to Amino sugar and nucleotide sugar metabolism. 4. By correlation analysis, we found that the RBC, ATPase, G6 PD, GPX, GR, methemoglobin, and EPO content were all on intimate terms with GSH metabolism, pentose phosphate pathway, Glycine, serine and threonine metabolism, Amino sugar and nucleotide sugar metabolism, and Alanine, aspartate and glutamate metabolism. And seven hemopenia biomarkers(L-Glutamic acid, Glutathione, Pyroglutamic acid, L-cysteine, D-Ribulose 5-phosphate, N-Acetylneuraminic acid, L-Glutamine) were found.In conclusion, GC-MS and LC-Q/TOF-MS both have advantages and disadvantages in metabolomics study. The mechanism and selection of hemopenia biomarkers by combining the two technologies. After the mice was built model with APH and CTX, Glutathione metabolism, Pentose phosphate pathway, Glycine, serine and threonine metabolism and Amino sugar and nucleotide sugar metabolism, alanine and glutamic acid and aspartic acid metabolism and their related metabolism were disturbed. Then it caused the decline of blood routine index, the abnormal of biochemical index, the clinical blood deficiency symptoms, and the atrophy of thymus tissue. After treated with UAS and its different processed products and Ejiao compound, the disturbed metabolism and the indexes were all restored to normal, and the blood deficiency symptoms disappeared. The mechanism of enriching blood of UAS and its different processed products was to intervene in blood deficiency by restoring Glutathione metabolism, Pentose phosphate pathway, Glycine, serine and threonine metabolism, Amino sugar and nucleotide sugar metabolism, alanine and glutamic acid and aspartic acid metabolism in the blood deficiency mice. Seven blood deficiency biomarkers were screened out.