Study On The Component Identification And Activity Evaluation Of Hedyotis Diffusa And Viburnum Setigerum

Hedyotis diffusa and Viburnum setigerum are traditional Chinese herbal medicines and have a long history.China has a wealth of Chinese medicinal resources^[1].However,natural medicines are not well-developed in terms of ingredient research,mechanism of action,etc.Therefore,it is of great significance to study the extraction,separation,and activity evaluation of natural medicine.In this thesis,silica gel column chromatography,ODS column,Sephadex LH-20column and high performance liquid chromatograph?HPLC?were used to separate and purify Hedyotis diffusa and Viburnum setigerum.Through MTT experiment,the neuroprotective activity of iridoid of Hedyotis diffusa on PC12 cells was demonstrated.And the hepatoprotective activity of ethyl acetate fraction of Viburnum setigerum on HepG2 cells was proved.This experiment provides scientific basis for the further mechanism research and application of the two traditional Chinese herbal medicines.This article compares the purification effects of iridoids from Hedyotis diffusa by different types of macroporous resin columns,polyamide columns,ODS columns,and Sephadex LH-20 columns.With Catalpol as the reference substance,the content was determined by chromogenic method.The results showed that the most suitable purification method was NKA-9 macroporous resin,and the content was 20.80%.By silica gel column chromatography and semipreparative high performance liquid chromatography?HPLC?,two iridoids were separated from the Hedyotis diffusa.6-O-methyldeacetylasperulosidic acid methyl ester and scandoside methyl ester were evaluated for the cytotoxicity and potential neuroprotective activity on PC12 cell lines which were exposed with H₂O₂ for 4h and they were determined by MTT method,with Edaravone as positive control drugs.The results showed that the survival rate of PC12cells could be significantly improved by the pretreatment of iridoids.Compared with the positive control drug edaravone,edaravone had a higher effect at low dose,while two iridoids were effective at middle and high doses.9 compounds were obtained from Viburnum setigerum by silica gel column chromatography and semipreparative high performance liquid chromatography,including taxifolin,?7?H,8??H?-4,4?,8?,9-Tetrahydroxy-3,3?-dimethoxy-7,9?-epoxylignan,?7?H,8?H,8??H?-4,4?,7??,9-Tetrahydroxy-3,3?-dimethoxy-7,9?-epoxylignan,?+?Catechin,Loganin,?-?Secoisolariciresinol,?7R,7?R,8R,8?S?-3,3?-Dimethoxy-7,7?-epoxylignane-4,4?,9,9?-tetraol,Alashinols G,Adoxoside.4 of them were evaluated for the cytotoxicity and potential neuroprotective activity on HepG2 cell lines which were exposed with H₂O₂ for 4h and they were determined by MTT method,with Edaravone as positive control drugs.The result showed that,?7?H,8??H?-4,4?,8?,9-Tetrahydroxy-3,3?-dimethoxy-7,9?-epoxylignan had cytotoxicity.The highest survival rate was 56.8%,and the lowest rate was 24.6%.At low dose Loganin had cytotoxicity.The lowest survival rate was 30%.At high dose,cell multiplication slightly changed.After two hydrogen quercetin pretreatment,the cell viability was not significantly changed,and the cell viability was only 31.2%.After a high dose of Loganin pretreatment,the cell viability increased significantly.When the concentration was 200?g/mL,the cell viability was 73.7%.The cell viability is dose-dependent with the concentration of Alashinols G.The cell viability increases first and then decreases.The cell viability was 51.2%when the concentration was 50?g/mL.