过氧化氢诱导血管内皮细胞凋亡与Caspase-3表达的实验研究

Hydrogen peroxide induced apoptosis in human vascular endothelial cells in vitro and expression of Caspase-3

目的:探讨过氧化氢对血管内皮细胞促凋亡作用及发生机制。方法:用0.1mmol/L、0.5mmol/L与1.0mmol/L等不同浓度的过氧化氢分别作用于对数生长期的内皮细胞,采用PI染色与Annexin V/PI染色后,进行流式细胞术检测内皮细胞的死亡率与凋亡率,并一步检测采用流式细胞术Caspase-3的水平表达。结果:过氧化氢0.1mmol/L、0.5mmol/L与1.0mmol/L组内皮细胞,凋亡率分别为18.6%±5.8%、32.8%±6.2%与42.1%±10.1%,而PI阳性率则为10.9%±4.1%、26.6%±5.3%与28.3%±4.6%,而Caspase-3的阳性表达率则为10.2%±2.1%、16.3%±3.2%和16.9%±3.9%。结论:过氧化氢可引起细胞凋亡,其中0.5mmol/L是诱导适宜浓度,且凋亡发生机制与Caspase-3激活相关。

Objective： To investigate the effects of hydrogen peroxide （H2O2） on apoptosis induction and its regulation mechanism in human vascular endothelial cells （HUVECs）. Methods： Cultured vascular endothelial cells （VECs） in logarithmic growth phase were treated with H2O2 of various concentrations, after PI and Annexin V/PI staining, flow cytometer were employed to detect and observe cell injury and expression of Caspase-3. Results： In H2O2 groups with different concentrations of 0. 1, 0. 5, and 1.0mmol/L respectively, H2O2 was able to induce apoptosis in HUVECs at the apoptosis rate of 18.6±5.8%, 32.8±6.2%, and 42.1± 10. 1%, furthermore, positive rates of PI staining were 10. 9±4. 1%, 26. 6±5. 3%, and 28. 3±4. 6%, correspondingly, positive rates of Caspase-3 expression 10. 2±2. 1%, 16. 3 ± 3. 2%, and 16. 9 ±3. 9%. Conclusion： H2O2 can induce apoptosis of vascular endothelial cells by regulation of concentration, and the appropriate concentration of H2O2 being around 0. 5mmol/L, moreover, activation of caspase-3 should involve apoptotic mechanism.