Metabonomic Study Of The Therapeutic Effects Of Siwutang On Blood Deficient Syndrome

Metabonomics, as an essential part of system biology, majors in quantitative measurement of metabolic response of living systems to pathophysiological stimuli or genetic modification. The metabolome represents all metabolites in biological cell, tissue or organism. Metabonomics concerns global response of living system especially end products of cellular processes. The rapid development of metabonomics and especially advances in the high-throughput and comprehensive research technologies provides new strategy in the analysis of traditional Chinese medicine. Blood deficient syndrome is often dignosed in the clinic of traditional Chinese medicine. We investigated the main factors which play important roles in the mechanism of pathogenesis and development of blood deficient syndrome for decade years. The effects of Siwutang on blood deficient syndrome involves in the recovering of stem and progenitor cells in bone marrow and circulating cells in blood, protection cell damage through DNA repair.To explore the relationship between blood deficient syndrome and the mechanism of Siwutang based on the involvement of changes in metabolic profiles, blood deficient mice induced by chronic bleeding. Thirty C57 mice were randomly divided into three groups including the control group and blood deficient model group without or with Siwutang treatment. The animals were sacrificed and blood, spleen, thymus and bone marrow were obtained. We investigated metabolic profiles of serum, aqueous and lipidic extracts of thymus, spleen and bone marrow using 1H nuclear magnetic resonance spectroscopy (NMR). The NMR spectra were integrated in segments of 0.01 ppm, normalized and then analyzed using SIMCA-P software to visualize the similarities and differences in metabolic profiles between these groups. Compared with control group, different levels of lactate, choline, glucose, taurine , alanine, LDL, glycerin, UFA, creatinine, acetoacetate, glutamate and β-hydroxybutyrate (β-HB) were observed in the model group. Siwutang could reverse these changes. The results suggested that the disorder of energy metabolism, damage of lymphocyte, and disorder of immune function were involved in blood deficient model, and the Siwutang could reverse these effects. Metabonomics may be a valuable tool in the pharmacological evaluation of Siwutang.UPLC-ESI-Q-TOF/MS was used to explore the relationship between blood deficient syndrome and the mechanism of Siwutang. Blood deficient mice were induced by radiation. Thirty-six C57 mice were randomly divided into three groups including the control group and blood deficient model group without or with Siwutang treatment. The animals were sacrificed on the 7th day after radiation and blood, spleen, thymus and bone marrow were obtained. The metabolic profiles of serum, aqueous and lipidic extracts of thymus, spleen were investigated using UPLC-ESI-Q-TOF/MS. The chromatogram were edited and analyzed by Markerlynx software.Lysophosphatide, hydroxybutyric acid, ceramide, phosphatidylinositol, carnitine, glucuronide were definited as the biomarkers of Siwutang to cure blood deficient syndrome. The results suggested that radiation could induce the disorder of the metabolism of lipoid and carbohydrate, also affect the synthesis of some amino acids, and the Siwutang could reverse these effects.A brief comparison of NMR and UPLC-MS has been proposed in the third section. The result indicated that more comprehensive metabolism information would be obtained if we unify the two different platforms. Thus, having a further understanding of the pathology and the physiological process would be possible.