黄连素激活核因子E2相关因子/血红素氧合酶-1增强内皮细胞抗过氧化氢损伤  

Protective effects of Berberine on H_2O_2-induced endothelial cell death through Nrf2/HO-1 pathway

作　　者：王臻[1] 丁慧[1] 杨瑞[1] Wang Zhen;Ding Hui;Yang Rui(Department of anesthesiology, Shaanxi Provincial People's Hospital, Xi’an 710068, China)

机构地区：[1]陕西省人民医院麻醉科,西安710068

出　　处：《中国体外循环杂志》2019年第1期41-47,共7页Chinese Journal of Extracorporeal Circulation

基　　金：陕西省科技研究发展计划(2012K13-02-23)

摘　　要：目的探讨核因子E2相关因子(Nrf2)/血红素氧合酶-1(HO-1)是否参与黄连素(BBR)介导人脐静脉内皮细胞(HUVECs)抗过氧化氢(H2O2)引起的凋亡以及潜在的机制。方法不同浓度BBR(5和10μmol/L)预处理细胞12 h后,应用H2O2(200μmol/L,4 h)构建细胞损伤模型,CCK-8法检测细胞活性; TUNEL标记凋亡细胞; DCFH-DA检测细胞活性氧(ROS)水平;免疫荧光法标记Nrf2细胞内的分布情况; Western blot法检测Nrf2/HO-1通路和凋亡相关蛋白含量。此外,应用干扰核糖核酸(siRNA)特异性阻断Nrf2/HO-1后,重复上述检测。结果相对于对照组,H2O2降低细胞活性,增加细胞凋亡及ROS水平(P <0.05)。不同浓度的BBR有效抑制上述变化,且呈"剂量依赖性"(P <0.05)。同时,BBR进一步促进H2O2导致的Nrf2核内移位及HO-1表达增加(P <0.05)。应用siRNA不仅可以显著抑制Nrf2/HO-1的活化,而且明显逆转BBR对HUVECs的保护作用(P <0.05)。结论 BBR通过激活Nrf2/HO-1通路,增强HUVECs清除ROS的能力,从而发挥抗H2O2损伤的作用。To investigate whether berberine(BBR)could protect human umbilical vein endothelial cells(HU.VECs)against H2O2-induced injury by activating nuclear factor erythroid 2-related factor 2/heme oxygenase-1(Nrf2/HO-1)sig.naling pathway.Methods After pretreated with BBR(5 and 10μmol/L)for 12 h,HUVECs cells were then insulted by H2 O2(200μmol/L)for additional 4 h.The cell viability was evaluated by CCK-8 analysis.The apoptotic cells were measured with TUNEL stain.ning.The cellular reactive oxygen species(ROS)was detected by DCFH-DA.The distribution of Nrf2 in cells was marked by immuno.fluorescence.Western blotting was used to analyze the proteins expression of Nrf2/HO-1 signaling pathway.Additionally,the detec.tions above were repeated after Nrf2 siRNA treatment.Results Compared with control group,H2 O2 decreased cell activity and in.creased apoptosis and ROS levels in HUVECs(P<0.05).Different concentrations of BBR(5 and 10μmol/L)effectively inhibited the changes above in a dose-dependent manner(P<0.05).Meanwhile,BBR further enhanced the translocation of Nrf2 from cyto.plasm to nucleus and the expression of HO-1,which was induced by H2O2 injury(P<0.05).Nrf2 siRNA not only significantly inhib.ited the activation of Nrf2/HO-1,but also obviously reversed the protective effects of BBR on HUVECs(P<0.05).Conclusion BBR promotes the ability of HUVECs to scavenge ROS by activating the Nrf2/HO-1 pathway in H2O2-induced injury.

关 键 词：黄连素 核因子E2相关因子/血红素氧合酶-1 人脐静脉内皮细胞 活性氧 凋亡 

分 类 号：R285.5[医药卫生—中药学]