The Correlation On Characterization Of Chemical Constituents Of Chaihu Guizhi Decoction And Its Pharmacodynamic Effcet Of Anti-liver Stagnation And Spleen Deficiency Stagnation And Infuenza Model

Objective:The chemical constituents of Chaihu Guizhi Decoction were studied and characterized.The active sites of Chaihu Guizhi Decoction of anti-influenza A virus（H₁N₁,H₃N₂）,Coxsackie virus（Cox-B5）,respiratory tract virus（RSV）,and herpes simplex virus（HSV-1）were studied by cytopathic effect（CPE）.In vivo mouse experiments were conducted to study the pharmacodynamic active components of Chaihu Guizhi Decoction in mice with liver stagnation and spleen exogenous model.Based on the chemical composition characterization and in vitro and in vivo pharmacodynamic tests,the related chemical substances group against influenza virus was screened.Methods:The chemical constituents of the compound of Chaihu Guizhi Decoction and each single-flavored decoction were determined by high performance liquid chromatography（HPLC）.Quantitative analysis of six main active ingredients of Chaihu Guizhi Decoction in the decoction of cinnamic acid,paeoniflorin,baicalin,baicalin,glycyrrhizin and glycyrrhizic acid;The chemical composition of Chaihu Guizhi Decoction decoction was qualitatively identified by quadrupole-electrostatic orbitrap high resolution mass spectrometry（UPLC-Q-Exactive/MS）;Based on in vitro cell antiviral experiments and in vivo liver stagnation and spleen exogenous model mice,the active sites of Chaihu Guizhi Decoction and the intervention of liver stagnation and spleen exogenous model mice were studied.PLS mathematical modeling analysis method was used to analyze the spectral effect of LC-MS fingerprint peak data and in vitro antiviral therapeutic index（TI）and liver spleen spleen exogenous model mice,and finally select the related drugs.Effective parts and chemical composition.Results:1.High performance liquid chromatography（HPLC）was used to analyze the fingerprint of Chaihu Guizhi Decoction,single-flavored medicinal decoction and extracts of different polar parts,and the medicinal materials were assigned to the compound in Chaihu Guizhi Decoction.At the same time,the content of six chemical components of cinnamic acid,paeoniflorin,baicalin,baicalin,glycyrrhizin and glycyrrhizic acid in Chaihu Guizhi Decoction was simultaneously determined by multi-wavelength high performance liquid phase method.2.The UPLC-Q-Exactive/MS technique was used to characterize the 53 components of Chaihu Guizhi Decoction.The structure of 16 compounds was determined according to the retention time and fragment ions of the standard.According to domestic and foreign literature reports and self-reports The Chaihu Guizhi Soup database inferred the possible structure of 37 compounds3.Based on cytopathic method（CPE）in vitro cell experiments,the effective active sites of Chaihu Guizhi Decoction and anti-viral drugs of different polar sites were studied.The experimental results showed that the compound,n-butanol and ethyl acetate parts of Chaihu Guizhi Decoction Influenza A virus H₁N₁ has a strong inhibitory effect（TI>10,has a killing effect）,compared with ribavirin-positive drug（TI=55.027）,Bupleurum Guizhi Tang compound（TI=68.7）has better inhibition The effect of petroleum ether and methylene chloride on Cox-B5 virus（TI>10,killing effect）,compared with ribavirin-positive drug（TI=17.396）,the effect is poor;The compound and n-butanol sites of Chaihu Guizhi Decoction have an inhibitory effect on RSV virus（TI>10,which has a killing effect）,and the effect is poor compared with the ribavirin-positive drug（TI=56.59）.4.Screening of pharmacodynamic activity of Chaihu Guizhi Decoction and different polar sites in mice with liver spleen deficiency and exogenous symptoms,through mouse body weight,rectal temperature,organ index,mortality,influenza A virus in lung tissue H1N1)mRNA relative expression,gastrin content and other comprehensive indicators,using least squares discriminant method to intervene in the mouse liver stagnation and spleen exogenous model of pharmacodynamic activity site,to determine the Chaihu Guizhi Decoction on liver spleen deficiency exogenous mouse model The treatment is best.5.UPLC-Q-Exactive/MS quadrupole-electrostatic field orbitrap high resolution mass spectrometry was used to establish the compound of Chaihu Guizhi Decoction,petroleum ether,methylene chloride,ethyl acetate,n-butanol and water after extraction.Fingerprint,combined with the in vitro antiviral therapeutic index and its comprehensive efficacy index for treating mice with liver stagnation and spleen exogenous model,through the"spectrum-effect"correlation study,it is clear that the decoction of Chaihu Guizhi Decoction and the extraction of different polar parts The characteristic chemical fingerprint peaks of anti-H1N1,Cox-B5,RSV virus and the intervention effect of liver stagnation and spleen exogenous model mice were further combined with UPLC-Q-Exactive/MS qualitative analysis results.The results showed that Chaihu Guizhi Decoction in water decoction Azelaic acid,Hexyl-β-sophoroside,glycyrrhizin,Codonopsis glucoside,9S,12S,13S-Trihydroxy-10E-octadecenoic acid,quercetin,and new glycyrrhizin have good effects on mice with liver spleen and spleen exogenous model.effect.For H₁N₁ virus,the results showed that with formononetin,protocatechuic acid,glycyrrhizin,baicalin,licorice flavonol,melamine A-7-O-β-D-glucuronic acid,gallic acid,There are some correlations between chemical components such as chrysin-6-C-α-L-pyran and baicalin.For Cox-B5 virus,the results showed that with scutellaria flavone II,cinnamic acid,baicalin,neoglycyrrhizin,paeoniflorinR1,formononetin,sedum,Liquiritigenin-4’-（1-2）-glucoside,Thechemicalcompositionof5,7,2’-Trihydroxyflavone is relevant.For RSV virus,the results were shown with Glycyrrhizin G,apigenin 6-C-glucoside-8-C-arabinosi,Schaffin,safrole,neoglycyrrhizin,6-glyphosin,and chrysin-6-C-There are correlations between chemical components such as glucose-8-C-arabinosides.