Investigation Of The Mechanism Of Bai-Shao And Zhi-Qiao Extracts In The Chaihu-Shu-Gan-Gan

Background Chaihu-Shu-Gan-San?CSGS?,the representative prescription of soothing liver and regulating vital energy,was originally documented in Jinyue-Quan-Shu which written by Zhang Jinyue.It involves seven commonly used Chinese herbs,i.e.the roots of Bupleurum chinense DC.?Chai-Hu?,the pericarps of Citrus reticulata Blanco.?Chen-Pi?,the roots of Paeonia lactiflora Pall.?Bai-Shao?,the fruits of Citrus aurantium L.?Zhi-Qiao?,the roots of Cyperus rotundus L.?Xiang-Fu?,the roots of Ligusticum chuanxiong Hort.?Chuan-Xiong?and the roots of Glycyrrhiza uralensis Fisch.?Gan-Cao?.Aim To investigate the material basis and mechanism of antidepressant effect of BS and ZQ from different angles and levels,which discover the quality control,reasonable clinical application,and secondary development foundation of CSGS.Method?1?The neuroprotective effects of CSGS,its subtracted formulas?CSGS without one of the herbs?extracts and its single herbs on the injured PC12 cells induced by corticosterone were investigated by morphological observation,MTT determination and LDH detection.?2?Tail suspension test?TST?and forced swimming test?FST?in mice were used to evaluate the antidepressant effect of Bai-Shao extracts and Zhi-Qiao aqueous extracts.?3?A rapid UPLC-Q-TOF/MS analysis was established and used to accurately online identify the chemical constituents of BS-60E preparation and ZQ aqueous extracts,respectively.?4?Cell metabonomics was firstly developed to evaluate the action of BS-60E against corticosterone-induced neurotoxicity on PC12 cells using UPLC-Q-TOF/MS.?5?In situ single-pass perfusion method was used to investigate the absorption and the concentrations in the perfusion were measured by UPLC.Results?1?The survival rate of PC12 cells exposed to corticosterone?200?mol/L?for48h decreased obviously,which the cell survival rate was 60.3%by MTT assay.However,the effect became weak when the CSGS,its subtracted formulas?CSGS without one of the herbs?extracts and its single herbs were added to the PC12 cells.?2?Administration of Bai-Shao extracts and Zhi-Qiao aqueous extracts significantly decreased the immobility time in TST and FST in mice,while BS-60E had best antidepressant effect.?3?26 compounds and 31 compounds were characterized in BS-60E preparation and ZQ aqueous extracts on the basis of retention behaviors,abundant MS and MS^E data,or by comparing with reference substances and literatures.?4?Both the MTT assay and the metabonomics results indicated that BS-60E had strong inhibition of the corticosterone-induced neurotoxicity on PC12 cells,where the pharmacological action of BS-60E was ascribed to the regulation on the glycerophospholipid metabolism and sphingolipid metabolism.?5?In perfusion mode,the P_effff of paeoniflorin,albiflorin,benzoyloxypaeoniflorin were 2.8,6.2,11.4(×10^-6cm·min^-1),whereas the P_effff of the extract were higher than the P_effff of the single component.Conclusion Combined with conventional pharmacological methods and metabolomics technology to explore the antidepressant effect of BS and ZQ from different angles and levels,in order to study the potential antidepressant chemical entities in CSGS and provide evidence for further development and application in clinic.