| Zhi-Zi-Chi decoction(ZZCD)comprised of Gardenia jasminoides Ellis and Semen sojae preparatum,derive from the classical composite herbal medicines formula Treatise on Febrile Diseases that has been used for the treatment of insomnia,anxiety,depression and neurasthenia and so on.In this Chinese medicine compound prescription group,Gardenia jasminoides Ellis(Rubiaceae)is an evergreen shrub and Semen sojae praeparatum(Chinese herbal name is “dan dou chi”)is a fermented food from Soybean,Sweet Wormwood Herb and Mulberry Leaf.As a commonly used traditional Chinese medicine,it has been recorded in detail in all visions of Chinese Pharmacopoeia.Currently,most of the studies were reported on active components analysis and pharmacological effects of single medicinal herbs including Gardenia jasminoides Ellis and Semen sojae praeparatum,there are few reports on ZZCD compound prescription for chemical constituents、pharmacological effects and significance of compatibility medicines.Firstly,we studied the neuroprotection of ZZCD on the injury of PC12 cells induced by glutamic acid(Glu).To experiment the effect on cell viability after adding different doses of Glu into PC12 cells,MTT assay was used to determine the cell viability,it was confirmed that the dose of 3 mmol/L Glu was chosen to establish the depression model.Subsequently,we experimented the protective effects of different concentration of ZZCD on the damaged PC12 cells,the results shown that the dose of 0.25,0.5 and 1 mg/mL had extremely protective effects on the damaged model.According to the study of LDH,GR,SOD and ROS,the PC12 cells induced by Glu could increase LDH release,significantly decrease the GR and SOD vitality,and enhance the level of ROS in PC12 cells.In addition,a series of ZZCD protection group could significantly decrease LDH release,increase the GR and SOD vitality,and reduce the level of ROS in PC12 cells.It was concluded that ZZCD had clear neuroprotection effects on the injury of PC12 cells induced by Glu.The results provided experimental data for subsequent study on chemical constituents analysis and clarification of compatibility significance.To study the chemical markers in Semen sojae praeparatum,a simple and effective quality fingerprint analysis coupled with chemometrics methods was developed for quality assessment of Semen sojae praeparatum based on the discussion of active components analysis in Gardenia jasminoides Ellis.And the chemical markers were analyzed qualitatively and quantitatively.Similarity analysis and hierarchical clusting analysis were applied to select the qualitative markers,which obviously influence the quality of Semen sojae praeparatum.21 chemicals were selected and characterized by LC-IT-TOF-MS.Subsequently,principal components analysis and orthogonal partial least squares discriminant analysis was conducted to select the quantitative markers of Semen sojae praeparatum.11 compounds including Daidzin,Glycitin,Genistin,Malonyl glycitin,Acetyl daidzin,Acetyl glycitin,Malonyl genistin,Daidzein,Glycitein,Acetyl genistin,Genistein with statistical significance were determined quantitatively.In order to study the chemical substance foundation of ZZCD in detail,the diversification of chemical composition influenced by compatibility was compared and recorded.A LC-IT-TOF-MS method was used to identify the constituents in Gardenia jasminoides Ellis,Semen sojae praeparatum and ZZCD,respectively.The compounds including crocin III,genipin,3,5-Di-O-caffeoyl-4-O-(3-hydroxy-3-methyl)glutaroy lquinic acid,malonyl glycitin,malonyl genistin,acetyl genistin were not detected in ZZCD,while Jasminoside A,Jasminoside I and two unknown compounds(m/z 611.1643,m/z 663.3227)were just detected in ZZCD.In addition,the changes of chemical components dissolution of ZZCD before and after compatibility were analyzed.It was concluded the optimum ratio of Gardenia jasminoides Ellis to Semen sojae praeparatum was 1to 1,which the contents dissolution of iridoid and isoflavone were the best. |
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