Investigation On The Spectrum-effect Relationships Of The Different Compatibility Of Active Fractions From Xiaoyaosan

Ethnopharmacology:Depression is a serious public mental disease. Xiaoyaosan (XYS) which is a valuable Traditional Chinese Medicine has been attracting more and more attention in recent years because of its effects for the treatment of depression in China. In our previous experiments, the antidepressant-like effects of XYS was also verified. Furthermore, our recent study indicated that three fractions of XYS were the active fractions which possessed the antidepressant-like effects. However, the bioactive components are usually not understood because of the complex chemical compositions of this medicine. In recent years, combination of chromatographic analysis with pharmacological effects to rapidly target the active constituents has been proposed and successfully applied to screen and analysis of the multiple bioactive compounds in herbal medicines.Aim of the study:Orthogonal design and factorial design were used to combine the active fractions from XYS into different compatibility groups. The antidepressant effect of compatibility groups could be different because of the distinction of constituent contents and reciprocal proportion. The spectrum-effect relationships of compatibility groups would be developed for analysis of active components in the XYS.Material and methods:Three active fractions (X7, X6, X5) from Xiaoyaosan were combined into different compatibility groups using L9(34) orthogonal design. The effects of the compatibility of active fractions on changes of behavior on stress model in mice were studied to investigate the contribution of each fraction on efficacy. The effects of compatibility groups on changes of sleep time on stress model in mice were also studied. Stress model in mice and HPLC-UV were used to screen the optimum extraction of active fractions. HPLC-UV was also used to establish the fingerprints of active fractions and investigation on process stability. A42factorial design was used to combine the active fractions (X7, X6) into different compatibility groups possessing equal constituents and different contents. Stress model in mice were selected to investigate the antidepressant effect of compatibility groups, the effects of compatibility groups on5-HIAA,5-HT and NE levels in the mouse brain regions of hippocampus were also examined in the forced swimming test. The spectrum-effect relationships of compatibility groups between chromatographic analysis (GC-MS and UPLC) and antidepressant-like effects were developed for analysis of active components in the XYS.Results:1. The contribution of X7and X6on efficacy were significant and X5was removed from compatibility groups. The compatibility group could decrease sleep time.2. Process7was used instead of process1(original extraction process) and was seen as the optimum extraction of active fractions, in addition, the process stability was well. The fingerprints of active fractions were established by HPLC-UV.3. The results indicated significant effects of compatibility groups on immobility time and5-HIAA,5-HT and NE levels in hippocampus in the mouse FST and TST. The antidepressant effects of compatibility groups possessing equal constituents and different contents were different. Antidepressant-like effects of X7were enhanced by X6and this interaction showed a significant synergistic antidepressant effect. The effects of compatibility group including X7and X6were significantly better than the effects of X7or X6, individually. The compatibility group (X7:92.6g/kg, X6:46.3g/kg) had the best anti-depression effect.4. The compatibility groups possessing equal constituents and different contents were analyzed by GC-MS and UPLC. The characteristic peaks were possessed collectively by the chromatographic fingerprints.5. Some compounds which might contribute to the antidepressant-like effects were selected using spectrum-effect relationships analysis. These compounds mainly came from Radix Bupleuri, Radix Angelicae Sinensis and Rhizoma Atractylodis Macrocephalae, while Radix Bupleuri, Radix Angelicae Sinensis and Rhizoma Atractylodis Macrocephalae are the main ingredients of the XYS formula. The activities of some of compounds were further confirmed by the literature, which suggest that the method of the combination of chromatographic analysis and pharmacological effect to study the spectrum-effect relationships based on compatibility of active fractions is suitable for hypothesis of active components of XYS.Conclusions:1. The antidepressant effects of compatibility groups were evaluated successfully.2. This work carried out chemical and pharmacological studies on XYS to elucidate the biologically active compounds. The results suggest that the strategy is useful for clarifying the pharmacology material basis of XYS.3. This work provides a model of combining chromatographic analysis and pharmacological effect based on compatibility of active fractions to search for bioactive components of TCM.