The protective effects of hyperoside on Ang II-mediated apoptosis of bEnd.3 cells and injury of blood-brain barrier model in vitro

Figures

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  Fig. 1. The effects of various concentrations of Hyp on the viability of bEnd.3 cells in vitro. To detect the safety of Hyp, cells were treated with different concentrations of Hyp (10, 20, 50, 100, 200, 500, 800 and 1000 μM) for 24 hr. The cell viability was measured by MTT assay. All the results were represented as mean ± standard deviation of the mean for six replicates from three independent experiments. #p < 0.05, ##p < 0.01 vs control

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  Fig. 2. The effects of various concentrations of Hyp against H2O2-induced damage in bEnd.3 cells in vitro. The cell viability was measured by MTT assay. All the results were represented as mean ± standard deviation of the mean for six replicates from three independent experiments. #p < 0.05, ##p < 0.01 vs control. *p < 0.05, **p < 0.01 vs H2O2 (600 μM) group

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  Fig. 3. The effects of different concentrations of Hyp on bEnd.3 cells apoptosis under Ang II-treatment in vitro (fluorescence microscopy, 200 × magnification). All nuclei were stained by DAPI with dark blue fluorescence. Apoptotic nuclei stained by TUNEL presented red fluorescence. The experiment was conducted in triplicate

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  Fig. 4. a Protection of deferent concentrations of Hyp against Ang II-induced apoptosis in bEnd.3 cells in vitro as detected by flow cytometry (V-FITC/PI) analysis. The data were analyzed by Flow Jo. b All the results were represented as mean ± standard deviation of the mean from three independent experiments. #p < 0.05, ##p < 0.01 vs control. *p < 0.05, **p < 0.01 vs Ang II (0.1 μM) group

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  Fig. 5. Hyp prevents Ang II-induced apoptosis in bEnd.3 cells via inhibiting mitochondrial and death receptor signaling pathways. a Western blot analysis showed the levels of apoptosis-related proteins in five groups. Actin levels were measured for the confirmation of equal amount of protein loading. The blots of Bcl-2 and Bax were cropped from the same gel. The blots of Caspase-3 and Cleaved Caspase-3 were cropped from the same gel. The blots of Caspase-8 and Cleaved Caspase-8 were cropped from the same gel. The blots of Caspase-9 and Cleaved Caspase-9 were cropped from the same gel. b The data were represented as mean ± standard deviation of the mean from three independent experiments. #p < 0.05, ##p < 0 .01 vs control. *p < 0 .05, **p < 0.01 vs Ang II (0.1 μM) group

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  Fig. 6. Hyp prevents Ang II-induced damage of BBB via regulating expression of the TJs and transcytosis-related proteins in Ang II-treated bEnd.3 cells. a Western blot analysis showed the levels of TJs and transcytosis-related proteins in five groups. Actin levels were measured for the confirmation of equal amount of protein loading. b The data were represented as mean ± standard deviation of the mean from three independent experiments. #p < 0.05, ##p < 0.01 vs control. *p < 0.05, **p < 0.01 vs Ang II (0.1 μM) group

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  Fig. 7. Protection of deferent concentrations of Hyp against Ang II-induced damage of BBB permeability in vitro as detected by determination of Na-F permeability. The data were represented as mean ± standard deviation of the mean from three independent experiments. #p < 0.05, ##p < 0.01 vs control. *p < 0.05, **p < 0.01 vs Ang II (0.1 μM) group