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目的观察蓝莓花色苷对过氧化氢(H_2O_2)所致血管内皮细胞损伤的保护作用及其机制。方法选取处于对数生长期的人静脉血管内皮细胞EVC-304,分别加入蓝莓花色苷10μg·ml~(-1)、20μg·ml~(-1)和40μg·ml~(-1),培养24h后再加入H_2O_2100μmol·L~(-1)作用12 h。MTT法检测细胞存活率,Hoechst 333258染色法观察EVC-304细胞凋亡形态,试剂盒测定超氧化物歧化酶(SOD)活性和丙二醛(MDA)含量,Western blot法检测细胞内胱天蛋白酶3(Caspase-3)的表达。结果同正常组相比,H_2O_2损伤组能够明显降低细胞存活率(P<0.01),细胞出现染色质固缩,核致密浓染,细胞核变小浓集的凋亡形态,SOD活性降低,MDA含量升高(P<0.01),Caspase-3蛋白表达呈升高趋势。与H_2O_2损伤组相比,蓝莓花色苷20μg·ml~(-1)和40μg·ml~(-1)组细胞存活率明显升高(P<0.01),凋亡细胞形态有所改善,SOD活性升高,MDA含量降低(P<0.01),细胞内Caspase-3表达下降。结论蓝莓花色苷对H_2O_2所致血管内皮细胞氧化损伤具有明显的保护作用,其机制可能与改善细胞内氧化应激水平相关。
Objective To observe the protective effect of anthocyanins from blueberry on vascular endothelial cell injury induced by hydrogen peroxide (H 2 O 2) and its mechanism. Methods Human logarithmic growth phase of human vascular endothelial cells, EVC-304, were treated with 100μg · ml -1, 20μg · ml -1 and 40μg · ml -1 of blueberry anthocyanin, After 24h, H_2O_2100μmol·L -1 was added for 12 hours. The cell viability was detected by MTT assay. The apoptotic morphology of EVC-304 cells was observed by Hoechst 333258 staining. The activity of superoxide dismutase (SOD) and the content of malondialdehyde (MDA) were determined by kit. The expression of intracellular caspases 3 (Caspase-3) expression. Results Compared with the normal group, the H 2 O 2 -treated group could significantly reduce the cell viability (P <0.01). The cells showed chromatin condensation, dense nuclear staining and apoptotic cell nuclei. The SOD activity decreased and MDA content (P <0.01), Caspase-3 protein expression increased. Compared with the H 2 O 2 -induced group, the viability of the cells in 20μg · ml -1 and 40μg · ml -1 groups was significantly increased (P <0.01), the morphology of apoptotic cells was improved, while the activity of SOD (P <0.01), while the expression of Caspase-3 decreased. Conclusion Anthocyanins of blueberry have a significant protective effect on H 2 O 2 -induced oxidative damage of vascular endothelial cells, which may be related to the improvement of intracellular oxidative stress.