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Objective To elucidate activity of baicalin against human cytomegalovirus ( HCMV) in vitro, and explore its effect on apoptosis of human embryo lung fibroblasts ( HEL ) infected with HCMV. Methods CCK-8 kit was used to determine the maximum tolerated dose ( MTC ) of HEL cell to baicalin while the anti-HCMV median efficacious concentration ( EC50 ) of baicalin was determined by standard plaque reduction method. After treatment with baicalin of different concentrations for 24, 48, 72 and 96 h, cell apoptosis and pro-caspase-3 expression was detected by flow cytometry and Western blotting, respectively. Results The MTC of baicalin was 20.6 μg?mL-1; The EC50 of anti-HCMV of baicalin was 16.13 μg?mL-1;The apoptosis rate increased gradually in the groups with low and high multiplicity HCMV infection at early time, showing significant dose-dependent manner. While the ratio of apoptotic cells was going to decrease in high multiplicity infection group 96 h after the infection. The expression of pro-caspase-3 was significantly higher in high-dose baicalin treatment group than in the infection control group ( P<0.05) . Conclusion Baicalin has a direct anti-HCMV effect in vitro. One of the mechanisms might be related with it inhibiting cell apoptosis and antagonizing activation of pro-caspase-3.
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Objective To elucidate activity of baicalin against human cytomegalovirus ( HCMV) in vitro, and explore its effect on apoptosis of human embryo lung fibroblasts ( HEL ) infected with HCMV. Methods CCK-8 kit was used to determine the maximum tolerated dose ( MTC ) of HEL cell to baicalin while the anti-HCMV median efficacious concentration ( EC50 ) of baicalin was determined by standard plaque reduction method. After treatment with baicalin of different concentrations for 24, 48, 72 and 96 h, cell apoptosis and pro-caspase-3 expression was detected by flow cytometry and Western blotting, respectively. Results The MTC of baicalin was 20.6 μg?mL-1; The EC50 of anti-HCMV of baicalin was 16.13 μg?mL-1;The apoptosis rate increased gradually in the groups with low and high multiplicity HCMV infection at early time, showing significant dose-dependent manner. While the ratio of apoptotic cells was going to decrease in high multiplicity infection group 96 h after the infection. The expression of pro-caspase-3 was significantly higher in high-dose baicalin treatment group than in the infection control group ( P<0.05) . Conclusion Baicalin has a direct anti-HCMV effect in vitro. One of the mechanisms might be related with it inhibiting cell apoptosis and antagonizing activation of pro-caspase-3.
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Kindling, a sub threshold chemical or electrical stimulation, increases seizure duration and enhances accompanied behavior until it reaches a sort of equilibrium state. The present study aimed to explore the effect of curcumin on the development of kindling in PTZ kindled rats and its role in apoptosis and neuronal damage. In a PTZ kindled Wistar rat model, different doses of curcumin (100, 200 and 300 mg/kg) were administrated orally one hour before the PTZ injections on alternate day during the whole kindling days. The following parameters were compared between control and experimental groups: the course of kindling, stages of seizures, Histopathological scoring of hippocampus, antioxidant parameters in the hippocampus, DNA fragmentation and caspase-3 expression in hippocampus, and neuron-specific enolase in the blood. One way ANOVA followed by Bonferroni post hoc analysis and Fischer’s Exact test were used for statistical analyses. PTZ, 30 mg/kg, induced kindling in rats after 32.0±1.4 days. Curcumin showed dose-dependent anti-seizure effect. Curcumin (300 mg/kg) significantly increased the latency to myoclonic jerks, clonic seizures as well as generalized tonic-clonic seizures, improved the seizure score and decreased the number of myoclonic jerks. PTZ kindling induced a significant neuronal injury, oxidative stress and apoptosis which were reversed by pretreatment with curcumin in a dose-dependent manner. Our study suggests that curcumin has a potential antiepileptogenic effect on kindling-induced epileptogenesis.
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Objective To investigate the effect of 5-azacytidine(5-AZA) on apoptosis of bone marrow mesenchymal stem cells(BMSCs).Methods BMSCs were isolated from bone marrow of mouse tibia and femur; the expression of MSC specific markers CD44 and CD90 in BMSCs was measured by immunofluorescence staining;BMSCs were cultured in vitro in the medium supplemented with 0,10 and 20 μmol/L 5-AZA for 48 hours.Cell apoptosis was measured with fluorescent labeled inhibitor of caspases (FLICA) apoptosis kit and 4',6-diamidino-2-phenylindole (DAPI) staining ;the expression of apoptosis-related proteins Annexin V and Caspase-3 in the treated BMSCs was detected by Western blot.Results In this study,BMSCs positively expressed MSC specific markers CD44 and CD90.DAPI staining and Caspase-3 staining both showed that 10 and 20 μmol/L 5-AZA markedly increased apoptotic rate of BMSCs;the apoptosis-positive rate in DAPI staining was (21.086 ± 2.601) ％,(34.467 ± 3.724) ％ and (46.512 ± 3.864) ％,the apoptosis-positive rate in Caspase-3 staining was (5.354 ± 0.735)％,(15.462 ± 2.385)％ and (28.190 ± 4.190)％ in the controls,10 and 20 μmol/L 5-AZA groups,and there were significant differences among the control group and 5-AZA treated groups(all P ＜0.01).Western blot assay showed that Annexin V and Caspase-3 were both markedly upregulated in 5-AZA treated cells;the relative level of Annexin V expression was(26.612 ±2.184)％,(42.873 ±4.313)％ and (50.056 ± 4.457) ％,the relative level of Caspase-3 expression was (19.231 ± 2.683) ％,(38.618 ± 5.385) ％ and(91.235 ± 7.116)％ in the controls,10 and 20 μmol/L 5-AZA groups,and there were significant differences among the control group and 5-AZA treated groups (all P ＜ 0.01).Conclusion The commonly used doses of 5-AZA can induce apoptosis of BMSCs.
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Objective: To evaluate the effect of curcumin on A549 tumor cell growth of subsets SP and NON-SP cells and the specific targets. Methods: The subsets SP and NON-SP cells were separated from A549 tumor strain of the human lung adenocarcinoma, and these cells were identified with soft agar clone formation test and tumorigenicity in vitro. Curcumin with different concentration (10, 20, 30, and 40 μmol/L) was used to deal with these cells. Using MTT method to make sure the inhibition of curcumin on the growth of SP and NON-SP cells. PCR was used to measure the expression of survivin and caspase-3. Results: Curcumin at different concentration showed the different inhibition on SP and NON-SP cells. Curcumin (30 μmol/L) showed more inhibition on SP cells than NON-SP cells with the lowest cell viability and was time-dependent. Curcumin could suppress the expression of survivin and promote the expression of caspase-3. The apoptosis degree of SP cell was clearly higher than that of NON-SP cell. Conclusion: Tumorigenicity of SP cell is higher than that of NON-SP cell. Curcumin induces the apoptosis of tumor cells through inhibiting the growth of SP cell, down-regulating the expression of survivin, and promoting the expression of caspase-3 gene.