Studies On The Chemical Constituents Of Patrinia Heterophylla Bunge. And Their Anti-tumor Activities

The SRB assay was used to find out the anti-tumor constituents from crude extraction which was extracted from Patrinia heterophylla Bunge.by 100%methanol. The results indicated the extraction of trichlormethane have the stronger cytotoxicity against to HepG2,SGC-7901 and HL-60 cell lines than other three parts of crude extraction,and IC₅₀were tested as:19.57±2.54μg/mL,44.04±5.04μg/mL and 17.91±0.88μg/mL.And four parts which were extracted respectively by petroleum benzine, trichlormethane,acetoacetate and,n-butanol hawe significant toxicity against HL-60 cell line,and their IC₅₀were tested as:33.72±5.82μg/mL,17.91±0.88μg/mL,31.49±3.70μg/mL and 40.23±7.39μg/mL.Especially,extraction of trichlormethane has the strongest effect to HL-60 ceilline.The further study was to separated activity compounds from extraction of trichlormethane,which has the strongest cytotoxicity.Finally,18 compounds were isolated from extract using silica gel column elution assay,and 11 compounds were identified by modern spectroscopy methods(NMR,MS,IR and UV),such as: friedelin,β-sitosterol,oleanolic acid,ursolic acid,hederagenin,daucosterol, canophyllol,scopoletin,oleanolic acid 3-O-a-L-arabinopyranoside,β-amyrin andα-myrin.There are seven triterpenoid compounds,two steroid compounds,one triterpenoid saponin and one coumarin compound.All of them were isolated from the plant for the first time besides oleanolic acid.SRB assay was used to determine cytotoxicities in vitro.The results indicatd that oleanolic acid,ursolic acid,hederagenin,scopoletin and oleanolic acid 3-O-a-L-arabinopyranoside have significant cytotoxicity against HepG2,SGC-7901 and HL-60 cell lines.The IC₅₀fluctuated from 4.56±0.48μg/mL to 18.07±0.05μg/mL. At the same time,it was found that the carboxyl group sited at C-28 is critical to cytotoxicity in triterpenoid which obtained in present study.The preliminary study of anti-tumor activity of hederagenin,a triterpenoid,was conducted by inverted microscope observation and typan blue exclusion in vitro.It was found that hederagenin can markedly inhibit cell proliferation in HL-60 cell line. Hoechst33258 stain assay and DNA ladder demonstrated that hederagenin can induce cell death by apoptosis pathway in HL-60 cell line.