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AIM:To study the apoptosis of hepatoma cells SMMC-7721induced by polysaccharide isolated from Ginkgo biloba seed.METHODS:Ginkgo biloba seed polysaccharide(GBSP)wasisolated by ethanol fractionation of Ginkgo biloba seed andpurified by Sephadex G-200 chromatography.The purity ofGBSP was verified by reaction with iodine-potassium iodideand ninhydrin and confirmed by UV spectrophotometer,cellulose acetate membrane electrophoresis and Sepharose4B gel filtration chromatography.The Scanning ElectronMicroscope(SEM)and Flow Cytometry(FCM)were used toexamine the SMMC-7721 ceils with and without GBSPtreatment at 500 mg/ml for 36 h.RESULTS:GBSP product obtained was of high purity withthe average molecular weight of 1.86×10~5.Quantitativeanalysis of SMMC-7721 cells in vitro with FCM showed thatthe percentages of G_2-M cells without and with GBSPtreatment were 17.01±1.28 % and 11.77±1.50%(P<0.05),the debris ratio of the cells were 0.46±0.12 % and 0.06±0.06 %(P<0.01),and the apoptosis ratio of cells was 3.84±0.55 %and 9.13±1.48 %(P<0.01)respectively.Following GBSPtreatment,microvilli of SMMC-7721 cells appeared thinnerand the number of spherical cells increased markedly.Mostsignificantly,the apoptosis bodies were formed on andaround the spherical cells treated with GBSP.CONCLUSION:GBSP could potentially induce the apoptosisof SMMC-7721 cells.
AIM: To study the apoptosis of hepatoma cells SMMC-7721 induced by polysaccharide isolated from Ginkgo biloba seed. METHODS: Ginkgo biloba seed polysaccharide (GBSP) wasisolated by ethanol fractionation of Ginkgo biloba seed andpurified by Sephadex G-200 chromatography. Purity of GBSP was verified by reaction with iodine-potassium iodide and ninhydrin and confirmed by UV spectrophotometer, cellulose acetate membrane electrophoresis and Sepharose 4B gel filtration chromatography. The Scanning Electron Microscope (SEM) and Flow Cytometry (FCM) were used toexamine the SMMC-7721 ceils with and without GBSP treatment at 500 mg / ml for 36 h.RESULTS: GBSP product obtained was of high purity with the average molecular weight of 1.86 × 10 ~ 5. Quantitative analysis of SMMC-7721 cells in vitro with FCM showed that the percentages of G_2-M cells without and with GBSPtreatment were 17.01 ± 1.28% and 11.77 ± 1.50%, respectively (P <0.05), the ratio of the debris were 0.46 ± 0.12% and 0.06 ± 0.06% (P <0.01), and the apoptosis ratio of c The number of ells was 3.84 ± 0.55% and 9.13 ± 1.48% (P <0.01) respectively. After GBFtreatment, microvilli of SMMC-7721 cells were thinnerand the number of spherical cells increased markedly. Hostsificantly, the apoptosis bodies were formed on and turned into the spherical cells treated with GBSP.CONCLUSION: GBSP could potentially induce the apoptosis of SMMC-7721 cells.