Study On Pharmacodynamics Of Antidepressant Effective Fraction From Xiaoyaosan

RATIONALES:After years study Xiaoyaosan had shown that it was a classics side of antidepressant, which had research by basic and clinical research. Our previous studies had demonstrated that the petroleum ether fraction of Xiaoyaosan had antidepressant activity. Depression is an emotional disorder caused by many factors, which has the characteristics of high morbidity, high mortality and morbidity. The diagnosis of clinical of depression at present is mainly use scale combined with clinical assessment, such as HAMD and SDS, is still a lack of objective, quantifiable depression diagnostic methods and standards, and the pathogenesis of depression not yet clear, complex potential indicators and lack of specificity.The efficacy of antidepressant drugs is also a lack of objective indicators. The dynamic effect of the drugs on the body is unclear. In recent years, scholars have conducted a lot of research, including neurotransmitters hypothesis. The small molecule compounds are found changed significantly with the development of metabolomics technology. Therefore, in this study behavioral indices as a benchmark, screening potential biomarkers from the neurotransmitter and small molecules. Then give positive drug screening and verification to obtain effect potential biomarkers. Through reserath the pharmacodynamics of the petroleum ether fraction of Xiaoyaosan, provide the basis for clinical.OBJECTIVE:1. Establish the HPLC and GC-MS detection method of serum and hippocampus. Through measured and analyzed find out the significantly different indicators, which was closed to depression.2. By giving the petroleum ether fraction of Xiaoyaosan, through the establishment of the measuring method of the quantitative determination of the effect index, verify the effect of indicators.3. To study the pharmacodynamic of the petroleum ether fraction of Xiaoyaosan according to the effect indicators. Supply a scientific basis for the next pharmacokinetic-pharmacodynamic research.METHOD:1. Establish the neurotransmitter in the serum and hippocampus HPLC-FD quantitative determination method; GC-MS metabonomics method is used to identify potential biomarkers and establish a potential biomarker of quantitative determination method.2. Seted up chronic unpredictable mild stress (CUMS) model, male adult SD rats were divide into model group (MS) and normal control group (NS). Collect serum on day0,6,9,12,15and22at the same time and conduct behavioral tests, collect hippocampus on day22. To find out effect indicators close to the depression, serum and hippocampus detect by HPLC and GC-MS. Screening and validation the effect effect indicators by principal component analysis and correlation analysis.3. Seted up chronic unpredictable mild stress (CUMS) model, male adult SD rats were divide into normal control group (NS), model group (MS), Xiaoyaosan group(XS) and venlafaxine group(YW). The experiment process was0-21day building while drug,22to42day building not to give medicine. Collect serum on day0,7,14,21,28,35and42at the same time and conduct behavioral tests. Serum was detect by HPLC and GC-MS. Acquired behavior and the effect of pre-screened indicators data, and validated. To study pharmacodynamics by verified the effect indicators.RESULTS:1. Establish determination HPLC-FD to detect the neurotransmitters in serum and GC-MS metabonomics method of quantitative analysis. They were conducted rigorous methodologies.2. In screening effect indicator of animal experiments, the behavior results showed that the model was successful. Found closely related to depression effect indexes have NE,5-HT, Glu, Gly, glucose, fructose, citric acid and hexadecanoic acid.3. In verify effect indicator of animal experiments, the behavior results also showed that the model was successful. The Xiaoyaosn group and venlafaxine group showed antidepressant effect after day21and the effect disappear after day35. Carried out the verification of the effect indicators, find out NE,5-HT, Gly and fructose were the final effect indicators.4. Pharmacodynamics study, the efficacy index to principal component analysis and data processing, using the DAS software carries on the analysis, get the efficacy of xiaoyao powder petroleum ether part and venlafaxine.Conclusions:In this study, screening and validation of endogenous small molecule compounds, which the body content of trends in modeling before and after administration were similar to behavior trend. It suggest that they chould be used as clinical diagnostic aid indicators of depression.