Study On The Effect And Mechanism Of Antidepressant Active Fraction Of Xiaoyaosan Based On Metabonomics

RATIONALES:Depression is a serious mental illness, which is called as the "emotional disorder". With the increasing pressure of people’s lives, the incidence of depression is the chance of getting higher and higher. It has been thank as the three black plague of the twenty-first century along cancer and AIDS by World Health Organization. Xiaoyaosan is from "Taiping huimin heji jufang" of Song Dynasty in the treatment of depression has been the validation of the clinical effects. However, the antidepressant effective fractions of the Xiaoyaosan is not clear. As an emerging science, metabonomics has been widely used in the medical field. The analytical methods like nuclear magnetic resonance (NMR), gas chromatography-mass spectrometry (GC-MS) and so on were used to analysis the change of endogenous metabolites from biological samples such as blood and urine and to analysis the body’s physiological and pathological changes. The systemic feature is consistent with the principle of TCM.OBJECTIVE:To explore the active fraction of Xiaoyaosan in producing antidepressant effect and to achieve the antidepressant mechanisms of the active fraction of Xiaoyaosan.METHODS:Xiaoyaosan through a series of separation processes, petroleum ether fraction (XY-A),30%ethanol fraction (XY-B),60%ethanol fraction (XY-C),95%ethanol fraction (XY-D) and water fraction (XY-E) were obtained. Through copying the chronic unpredictable stress model, the healthy adult male Sprague-Dawley (SD) rats were divided into seven groups:control group, chronic unpredictable stress model group, petroleum ether fraction group,30%ethanol fraction group,60%ethanol fraction group,95%ethanol fraction group and water fraction group. Urine samples were collected in21days. Plasma samples were collected on22day. The NMR and GC-MS analysis techniques were applied to determine the biological samples. The SIMCA-P software for the multivariate analysis was used to analysis the active fraction of Xiaoyaosan in producing antidepressant effect and the antidepressant mechanisms of the active fraction of Xiaoyaosan.RESULTS:(1) The results of metabolic studying:The data obtained from the two analysis techniques through multivariate statistical analysis showed the control group and model group can be clearly separated, which showed the model was successful. With the corresponding loading plot and independent sample T test, the potential biomarkers were found. The multivariate statistical analysis results for control group, model group and different fractions of Xiaoyaosan show that the different intervention effect of different fractions of Xiaoyaosan to the change of metabolites. Compared with the other fractions of Xiaoyaosan, the petroleum ether fraction of Xiaoyaosan was much closer to the control group and was the farthest away from the model group, indicating its significant antidepressant effect. The different fractions of Xiaoyaosan can regulate potential biomarkers tend to the normal control group, while the petroleum ether fraction of Xiaoyaosan can regulate most potential biomarker levels tend to control group.(2) Antidepressant mechanism research of active fraction of Xiaoyaosan:the petroleum ether fraction of Xiaoyaosan abate or cancel the stimulus factors on the function of organism, so that the metabolic spectrum of CUMS rats tend to the control group. Its mechanism of action may be related to potential biomarkers likes succinic acid, lactic acid and glycine. They can regulate the aspect of energy metabolism, nerve system and so on.CONCLUSION:The article copied the chronic unpredictable stress model successfully and analysis the collection of biological samples (urine, plasma) through metabolomics. The results identified the active fraction of Xiaoyaosan in producing antidepression and antidepressant mechanism of active fraction of Xiaoyaosan. The antidepressant effect of petroleum ether fraction is best in five different polar fractions of Xiaoyaosan, This will provide the new ideas for the study of material basis for the efficacy of Xiaoyaosan and provide the basis for the new drug research.