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Objective To investigate the effects of extract of Ginkgo biloba leaves EGb761 on 1-methy-l 4-phenylpyridium (MPP+)-induced injury in human neuroblastoma SH-SY5Y cells; To discuss its mechanism of action.Methods Cell culture method was used and SH-SY5Y cell damage model was induced with different concentrations of MPP+ to build Parkinson’s disease model in vitro. The experiment was divided into control group, MPP+ model group, low-, medium-, and high-dose EGb761 groups. The survival rate was determined by MTT assay, and the apoptotic rate was detected by flow cytometry according to AnnexinV apoptosis detection kit. The cell morphology was observed by inverted microscope. NO content in SH-SY5Y cells was detected by Nitric acid reduction method.Results Compared with the control group, the survival rate of SH-SY5Y cells decreased and the apoptotic rate and NO content increased in the model group (P<0.05); Compared with the model group, the survival rate of SH-SY5Y cells increased and the apoptotic rate and NO content decreased in the low-, medium- and high-dose EGb761 groups (P<0.05).Conclusion EGb761 can protect MPP+-induced SH-SY5Y cell from damage by the inhibition of the content of NO free radical.
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OBJECTIVE:To investigate inhibitory effects of matrine on the proliferation of human bladder cancer BIU-87 cells and its mechanism. METHODS:The cell viability was detected by MTT assay and inhibitory rate of cell proliferation was calculat-ed after human bladder cancer BIU-87 cells were treated with 0(negative control),0.5,1.0,2.0 and 4.0 mg/ml matrine for 24,48 and 72 h,respectively. After treated with 0 (negative control),0.5,1.0,2.0 and 4.0 mg/ml matrine for 48 h,the cell cycle and apoptotic rate were detected by flow cytometry;the expression of Survivin,Caspase-3 and Caspase-7 protein were detected by Western blot assay. RESULTS:Compared with negative control,the proliferation of BIU-87 cells were significantly inhibited after incubated with 1.0-4.0 mg/ml matrine for 24,48 and 72 h(P<0.05 or P<0.01),and inhibitory rate of cell proliferation increased in concentration and time-dependant manner;after treated for 48 h,the percentage of G0/G1 phase cells and apoptotic rate in-creased,while the percentage of cells at S phase and G2/M phase were decreased (P<0.05 or P<0.01);the expression of Cas-pase-3 and Caspase-7 protein increased,while the expression of survivin protein decreased after incubated with 0.5-4.0 mg/ml ma-trine for 48 h. CONCLUSIONS:Matrine can inhibit the proliferation of human bladder cancer BIU-87 cells,block the cell cycle and induce apoptosis;its mechanism may be related to the expression regulation of Survivin,Caspase-3 and Caspase-7.
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Objective To investigate the effects of ligustilide on the proliferation of human lung adenocarcinoma A549 cells and its mechanism. Methods CCK-8 method was used to detect the effects of ligustilide on activity of A549 cells. Apoptosis rate was measured by TUNEL. Nuclear morphological changes of A549 cells were observed by fluorescence microscope after staining by Hoechst 33258. ELISA was used to detect the VEGF levels after incubation with ligustilide. Western blot was used to detect the expression of NF-κB, Bcl-2, Bax and the protein expression of phosphorylation of JNK. Results A549 cell activity was significantly inhibited after incubated with 15, 30, 60, 120, 180μmol/L ligustilide for 12, 24, 48 h (P<0.05, P<0.01, P<0.001) in a dose and time-dependent manner (P<0.05, P<0.01);Apoptosis rate increased by 30, 60, 120 μmol/L ligustilide for 12, 24, 48 h in a dose and time-dependent manner;A549 cells treated with ligustilide for 48 h appeared the apoptosis forms, including nuclear shrinkage, beating, strong blue fluorescence staining, and the chromatin divided producing apoptotic body;30, 60, 120μmol/L ligustilide increased the expression of p65 subunit of NF-κB protein in the nuclei and the phosphorylation levels of JNK protein, reduce the expression of suppression apoptosis protein Bcl-2, and raised the expression of pro-apoptotic protein Bax (P<0.05, P<0.01, P<0.001). Conclusion By inhibiting the expression of VEGF, ligustilide can inhibit the formation of tumor angiogenesis. By regulating the expression of NF-κB and the phosphorylation levels of JNK protein and reducing the ratio of Bcl-2/Bax, ligustilide can promote tumor cell apoptosis.
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Objective To investigate the proliferation inhibitory effect and mechanism of Ginkgo Biloba extract (EGB) on non-small cell lung cancer (NSCLC) PC-9 cells. Methods Concentrations of EGB were set as 70 mg/L, 105 mg/L, 140 mg/L, 210 mg/L, and 280 mg/L, and were used to culture PC-9 cells in vitro for 24 h, 48 h, and 72 h. Tetramethylazo thiazole blue staining (MTT) method was applied to detect cell inhibition rate. Hoechst33258 fluorescent staining was employed to observe the nucleus. SOD activity and MDA content were detected by ELISA. The protein expressions of Caspase-3, Caspase-9, Cyt-C, and AIF were detected by Western blot. Results After incubated for 24 h, 48 h, and 72 h, EGB inhibited the proliferation of PC-9 cells, IC50 was 195.45 mg/L, 179.63 mg/L, 142.23 mg/L, respectively). 105 mg/L, 140 mg/L, 210 mg/L EGB could induce apoptosis of PC-9 cells, cause the nucleus pycnosis, produce apoptotic bodies, improve SOD activity and decrease MDA content (P<0.05, P<0.01, P<0.001). Western blot results showed that, compared with the control group, EGB can obviously increased the protein expressions of Caspase-3, Caspase-9, Cyt-C, and AIF, with statistical significance (P<0.05, P<0.01, P<0.001). Conclusion EGB can effectively restrain proliferation of non-small cell lung cancer PC-9 cells, the mechanism may be realized by inducing PC-9 cell apoptosis through pathway of mitochondria apoptosis, reducing oxidation, and clearing free radicals.