| Part IEffect of Hedyotic diffusa on apoptosis,proliferation and cell's cycle of U87glioma cells in vitroObjective:To investigate the effect and the possible mechanism of apoptosis induction by hedyotic diffusa (HD) on human glioma U87cells.Method:U87cells were cultured for24and72hours in medium which contained hedyotic diffusa with different concentrations, the methyl thiazolyl tetrazolium (MTT) assay was used to detect the survival rate, the apoptosis of the cells was examined by flow cytometry, the cell's cycle was examined by flow cytometry.Results:The cells proliferation was obviously inhibited by hedyotic diffusa and showed a dose dependent manner. The apoptotic rates of the4and8mg/mlHD groups were17.31±1.36)%,41.23+0.72%respectively, all significantly higher than that of the0mg/ml HD group(P<0.05).The result of cell's cycle showed HD could effect the U87cell's cycle distribution。 When compared w ith o ther groups, cells in stage of S increased in HD4mg/ml group and cells in stage of G2/M increased in HD8mg/ml, the rate of G0/G1 showed a dose-dependent decreased in these two group (P<0.05).Conclusion:Hedytotic diffusa could induce glioma U87cells apoptosis.After analysis of result of cell's cycl, We considered that HD could cause phage arrest and induce the apoptosis of U87glioma cells. Part IIThe study on the mechanism of suppression effect of Hedyotic diffusa on human glioma cells growth in vitroObjective:To investigate the possible mechanism of of suppression effect of Hedyotic diffusa on human glioma U87cells growth in vitroMethods:U87cells were cultured for24hours in medium which contained hedyotic diffusa with different concentrations(0,4,8mg/ml),the apoptosis of the cells and collapse of mitochondrial membrane potential were examined by flow cytometry,the change of Bcl-2and Bax proteins levels were measured by immunoblot.Results:The protein expression of caspase-3was increased dose-dependently.And the proteins expression of Bax was increased and Bcl-2-to-Bax ratio was reduced dose-dependently. The JC-1-green-bright rates of cells treated with4,8mg/ml HD were (11.90+0.20)%,(34.17+2.79)%respectively,significantly higher than untreated U87cells (P<0.05)Conclusion Hedytotic diffusa induced glioma U87cells apoptosis via mitochondrion pathway, then suppress human glioma U87cells growth. Part ⅢEffect of Hedyotic diffusa on U87glioma cells growth in vitroObjective To investigate the effect and the possible mechanism of hedyotic diffusa (HD) on human glioma U87cells.Method U87human glioma was inoculated into ICR mice to establish a human glioma xenograft glioma model. The tumor were treated by intratmoral injection with oldenlandia diffusa in different dosages. The negative control group was given saline, growth of tumor was observed, volume inhibition rates were calcutated and compared among every group. Study the change of HE staining and compare the side effect among every group.Results Volume inhibition rates of treated group were38%and42%respectively, there were significant difference between treated group and control group(P<0.05).HD injection group and HD injection add saline group took effect more slowly than cisplatin group, showed obvious suppression to tumor growth after treated with HD for21days. HE staining showed large necrosis area, inflammatory cell infiltration was found at the edge of necrosis, hemangiectasis and congestion varied to various degrees. There were no side effect after treated with HD.Conclusion Hedytotic diffusa could inhibitgrowth of tumor and no side effect. |
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