The Pharmacodynamic And Pharmacokinetic Studies On Antidepressant Effect Of Chaihu-Shugan-San Extract And Its Constituents In Rat Models Of Depression

Object:Use ultra-high performance liquid chromatography-Photo diode array (UPLC-PDA) to determine the the content of the active ingredients①To develop a simple, rapid and sensitive Ultra performance liquid chromatography coupled with photodiode array (UPLC-PDA) detection method for simultaneous determination of twelve compounds in Chaihu-Shugan-San:gallic acid, oxypaeoniflora, albiflorin, liquiritin, ferulic acid (FA), narirutin, naringin, hesperidin, neohesperidin, meranzin hydrate (MH), liquiritigenin, isoliquiritigenin,②To develop a liquid chromatography-tandem mass spectrometry (LC-MS) method for the determination of FA and MH in plasmas of patients with functional dyspepsia after oral administration of Chaihu-Shugan-San decoction.③To investigated the effect of CSS, FA and MH on gastrointestinal motility in rats, partly elucidate the effective substance of CSS treating functional dyspepsia.④To develop a Ultra performance liquid chromatography coupled with photodiode array (UPLC-PDA) detection method to explore the pharmacokinetics of FA and MH in plasmas of patients with functional dyspepsia after oral administration of CSS decoction.Methods:(1) The quantitative analysis of the sixteen active constituents in CSSThe analysis was performance on UPLC-PDA system with BEH C18column (100*2.1mm id,1.7μM). The mobile phase comprising acetonitrile (A) and0.5%acetic acid (B) was used to elute the targets in gradient elution mode. Flow rate and detection wavelength were set at0.5ml/min and284nm. The sixteen constituents in CSS including gallic acid, oxypaeoniflorin, albiflorin, paeoniflorin, liquiritin, FA, narirutin, naringin, hesperidin, neohesperidin, MH, liquiritigenin, quercimelin, benzoylpaeoniflorin, isoliquiritigenin and formononetin were quantitative determinate, and the limit of determination (LOD), precision, recovery, sample stability and reproducibility of the sixteen constituents were examined in this study.(2) Use the UPLC-PDA method to analyze the active ingredient in rats plasma after oral administration of CSS decoction.(3) To study the antidepression effects of CSS and its components on the wild chronic stress rat model,110Sprague-Dawley rats was randomly divided into11groups:the NC group、the MC group、the FC group、the CSS group、and seven components group,each group for10rats(4) To investigated the effect of CSS, FA on antidepression effects in rats, partly elucidate the effective substance of CSS treating depression. Then comparatively study on the bioethnopharmaceutical analytical pharmacology between CSS decoction and FA.Results:(1) Compared with the normal control group, Rats in model group, which subjected to a combination of a series of daily CUMS and housing singly for2-4week showed a significant decrease of body weight gain(P<0.01), a significant decrease in the number of crossings, rearings and grooming, a significant increase in the time for staying in the central square in open-field test(P<0.01), and a significant reduction in totalliquid consumption,sucrose consumption and the percentage of sucrosepreference(P<0.05or P<0.01).(2) After2-week treatment, compared with the model group, the number of body weight gain, crossings, rearings and grooming, sucroseconsumption and the percentage of sucrose preference in the rats of the CHSGS group were significantly increased, the time for staying in the central square were significantly decreased (P<0.05or P<0.01); The rats in the component Ⅰ group, the component Ⅱ group and the fluoxetine group did not show significant differences in body weight.Conclusions:(1) UPLC-PDA can determine the paeoniflorin in a effective way.(2) After the establishment of chronic unpredictable mild stress bioactive component which rat plasma can absorb. This components may be an effective ingredients to treatment of RA by Radix Paeoniae Alba,and it may be the efficacy material basis of CSS to treat of Depression.