依达拉奉对过氧化氢诱导血管内皮细胞Nrf2/HO-1表达的影响

Effect of edaravone on the expression of Nrf2/HO-1 by H_2O_2 in vascular endothelial cells

目的探讨依达拉奉对过氧化氢诱导血管内皮细胞Nrf2/HO-1表达的影响。方法将对数生长期的血管内皮细分为对照组、依达拉奉组、H_2O_2组、依达拉奉+H_2O_2组、H_2O_2+N-乙酰半胱氨酸(NAC)组,用不同条件的培养基培养48 h后用噻唑蓝(MTT)法检测细胞增殖,活性氧(ROS)检测试剂盒检测细胞内ROS,Western blot方法检测Nrf2/HO-1蛋白表达,PCR法检测Nrf2/HO-1 mRNA表达。结果依达拉奉促进血管内皮细胞的增殖,拮抗H_2O_2诱导血管内皮细胞增殖抑制作用。与对照组比较,依达拉奉组ROS无明显增高(P>0.05),Nrf2/HO-1蛋白和mRNA表达明显增高(P<0.05);而H_2O_2组ROS明显增高(P<0.05),Nrf2/HO-1蛋白和mRNA表达明显下降(P<0.05)。与H_2O_2组比较,依达拉奉+H_2O_2组和H_2O_2+NAC组ROS明显降低(P<0.05),Nrf2/HO-1蛋白和mRNA表达明显增高(P<0.05)。结论 H_2O_2下调血管内皮细胞Nrf2/HO-1表达,而依达拉奉通过清除细胞内ROS,拮抗H_2O_2下调细胞Nrf2/HO-1表达,从而抑制H_2O_2导致血管内皮细胞损伤。

AIM To investigate the effects of edaravone on the expression of Nrf2/HO-1 induced by H202 in vascular endothelial ceils. METHODS The vascular endothelial cells used in this study were divided into 5 groups： control group, edaravone group, H202 group, edaravone ＋ H202 group and HzO2 ＋ N-acetylcysteine （NAC） group. The viability of endothelial cells was tested by MTT. The intracellular reactive oxygen species （ROS） production was detected by reactive oxygen species assay kit. The protein expression of Nrf2/HO-1 was de- tected by Western blotting and the expression of Nrf2/HO-1 mRNA was detected by PCR. RESULTS Edaravone promoted the proliferation of endothelial ceils and decreased the inhibition induced by n202. Compared with the control group, the ROS production remained at same level with the control group and the expression of Nrf2/HO-1 increased significantly in edaravone group （P 〈 0.05） , while the ROS production increased and the expression of Nrf2/HO-1 decreased significantly in H202 group. Compared with H202 group, the ROS production decreased sig- nificantly and the expression of Nrf2/HO-1 increased in edaravone ＋ H202 group and H202 ＋ NAC group（ P 〈 0.05）. CONCLUSION Edaravone can increase the expression of Nrf2/HO-1, which is down-regulated by H202, through clearing intracellular ROS and protect vascular endothelial ceils injury induced by H202.