Investigation On The Anti-depression Effect And Mechanism Of Compound Chaigui Fang

Rationales:Depression is known as a complicated etiological disease, accompanied by multi-system dysfunction. It behaves high incidence as well as relapse rate and even leads to suicidal thoughts or behaviors, which is quite harmful to human’s health. Currently, the synthetic antidepressants have owned the major market of anti-depressants, and few breakthroughs about new anti-depressants have been made in recent years. Nowadays, the traditional Chinese medicine has been proved to be efficient in clinic use. However, its mechanism and efficacy haven’t been clearly known. According to the multi-target character, developing new antidepressants from traditional Chinese medicine has attracted a lot of attention.Previously, we studied on chemical and pharmacological characters of antidepressant components in Xiaoyaosan, and indicated that the low polar components played an important role. Then we got a new prescription including radix, atractylodes, angelica, ginger, licorice and mint herb after removing the poria and peony because their little contribution to the low polar components in Xiaoyaosan, accompanied with the supercritical CO2 extraction process. This project aims to obtain more economical production for solvent extracted components, and screen and then verify the prescription from the pharmacological perspective, and then explore its mechanism of action based on the previous studies.Objective:To screen the proper prescription and antidepressant components and verify its effectiveness, the prescription of Xiaoyaosan, extraction concentration and purification process were investigated from effectiveness based on the ethanol solvent. To elucidate the antidepressant pathways and mechanisms of the components, it will be helpful for the discovery of new antidepressant using metabolomics and neuroendocrine hormone technologies.Methods:Behavioral despair in mice model (the tail suspension test, the forced swimming test) using the immobility time as evaluation index was conducted for screening the prescription composition, extraction concentration and purification process, and the 75% ethanol-ethyl acetate extracts of Compound Chaigui Fang (FFCGF-EE) was identified for drug development. The chronic unpredictable mild stress (CUMS) of depressive rat model was used to verify the antidepressant effect of FFCGF-EE from the various behaviors; the 1H NMR metabolomics of CUMS rat serum combined with multivariate statistical analysis was used to analyze the change of metabolites, identify the potential biomarkers, evaluate the effectiveness of FFCGF-EE and investigate the antidepressant mechanism. The contents of hormones of CUMS rat serum that related with the hypothalamus-pituitary-adrenal cortical axis (HPAA), the hypothalamus-pituitary-thyroid anis (HPTA) and the hypothalamus-pituitary-gonadal axis (HPGA), such as adrenocoticotropic hormone (ATCH), corticosterone (CORT), thyroid stimulating hormone (TSH), three iodine thyroid glycine (T3), thyroxine (T4) and testosterone, were analyzed using enzyme linked immunosorbent assay (ELISA) after FFCGF-EE treatment, and the correlation analysis was studied between biomarkers and hormones.Results:1. According to effectiveness, the Compound Chaigui Fang that removed the "Radix Paeoniae Alba" and "Tuckahoe" from Xiaoyaosan, and the 75% ethanol-ethy acetate extract were identified. Based on the behaviors of CUMS rats, such as body weight, appearance, open field test, sugar preference and elevated plus-maze test, it was confirmed that antidepressant effects of the moderate and high doses of FFCGF-EE were similar to Xiaoyaosan prescription, and closed to the positive drug.2. Based on 1H NMR metabolomics analysis, the rat serum metabolic spectra after CUMS exposure and FFCGF-EE treatment were analyzed, and six biomarkers including lactate, leucine and choline etc. were found, and the levels of biomarkers were close to the normal after drug treatment, and the antidepressant mechanism of FFCGF-EE may be related with the regulation of glycometabolism metabolism, amino acid metabolism and choline metabolism.3. The ELISA results of different treated groups on rat serum indicated that the levels of ACTH, CORT and T3 in CUMS group significantly increased (P<0.01, 0.05), combined with the significantly decreased levels of TSH and T (P<0.01,0.05), which could be regulated to the normal levels after FFCGF-EE treatment. The FFCGF-EE could exert the regulation of neuroendocrine function.Conclusion:The study indicated that FFCGF-EE exerts definite antidepressant effect and could replace the Xiaoyaosan prescription to further development. The antidepressant mechanisms of FFCGF-EE were related with the regulation of serum endogenous metabolites and HPAA, HPTA, HPGA.