The Preliminary Studies On Active Components From Fructus Xanthii Inhibiting Breast Carcinoma Cells And Herpes Simplex Virus 1

Fructus Xanthii,called Cang-Erzi in Chinese Pin Yin,is the dried ripe fruit of Xanthium strumarium L.(Asteraceae).It has been commonly used as a traditional Chinese medicine for treatment of sinusitis and rheumatic headache,which is recorded in the pharmacopoeia of People’s Republic of China and widely distributed in China.Fructus Xanthii contains various chemical constituents and the modern pharmacological studies have validated that Fructus Xanthii possessess anti-tumor,anti-virus,anti-bacterial,anti-inflammatory,analgesic,anti-oxidant,anti-diabetic and other properties.The anti-tumor and anti-virus components were attempted to isolate and purify from raw materials Fructus Xanthii using breast cancer cell line MCF-7 and herpes simplex virus type1(HSV-1).The specific experiments process and results can be seen as below:1.Anti-tumor(MCF-1)experimentsInhibiting proliferation of MCF-7 cells was used as the activity tracking index.The constituents were isolated and purified by organic solvent extraction,silica gel column chromatography,semi-preparative high performance liquid chromatography(HPLC)and gel column chromatography methods.In the process of anti-cancer activity screening,MTT assay was employed to detect the effect of different components on the cell viability rate of MCF-7 cells.The anti-tumor activity was determined by IC50 values.Eventually,Annexin V-FITC/PI double staining and flow cytometry analysis were adopted to discover the relationship between the compound and apoptosis of MCF-7 cells.EtOAC fraction,nBuOH fraction and H2O fraction of Fructus Xanthii were obtained after extracted by organic solvents.After the activity screening,the EtOAC fraction was identified as the active component.Twelve components were obtained after separation by silica gel column chromatography,and EFd was the active component.After separation by semi-preparative HPLC,four components were obtained,in which EFd-3 was the active fraction.Finally,gel column chromatography was used to obtain the active monomer compound EFd-3-1,and the IC50 values is 21.54μg/ml.EFd-3-1 can induce apoptosis of MCF-7 cells in a concentration-dependent manner.2.Anti-virus(HSV-1)experimentsSupressing the cytopathic effect(CPE)of Vero cells induced by HSV-1 was used as the activity tracking index.The constituents were isolated and purified by organic solvent extraction,silica gel column chromatography and semi-preparative high performance liquid chromatography(HPLC)methods.During screening anti-virus activity,CPE inhibition assay was performed to detect the effect of different components on cytopathy of Vero cells.The anti-virus activity was determined by SI values.EtOAC extract,nBuOH extract and aqueous extract were obtained after extracted with organic solvent,and EtOAC extract was identified as the active component.After separation by silica gel column chromatography,twelve components were obtained,where EFe was the active ingredient.After separation by semi-preparative HPLC,three components were obtained,in which EFe-2 was the active fraction,and the SI value is 1.81.