丁苯酞对S-亚硝基化谷胱甘肽诱导脑微血管内皮细胞损伤的保护作用及机制研究  

Protective effects of Butylphthalide on S-nitrosoglutathione-induced brain microvascular endothelial cells injury and potential mechanisms

作　　者：严洁萍[1] 章水均[2] 张国兵[1] 胡颖[1] 罗丹[1] 江红娟[1] 李敏静 YAN Jieping;ZHANG Shuijun;ZHANG Guobing;HU Ying;LUO Dan;JIANG Hongjuan;LI Minjing(Department of Pharmacy,Zhejiang Provincial People's Hospital,People's Hospital of Hangzhou Medical College,Hangzhou 310014,Zhejiang,China;Department of Orthopedics,Zhejiang Provincial People's Hospital,People's Hospital of Hangzhou Medical College,Hangzhou 310014,Zhejiang,China;Department of Respiratory,The Second Affiliated Hospital of Zhejiang Chinese Medical University,Hangzhou 310053,Zhejiang,China)

机构地区：[1]浙江省人民医院杭州医学院附属人民医院药学部,浙江杭州310014 [2]浙江省人民医院杭州医学院附属人民医院骨科,浙江杭州310014 [3]浙江中医药大学附属第二医院呼吸内科,浙江杭州310053

出　　处：《中国临床药理学与治疗学》2019年第5期517-522,共6页Chinese Journal of Clinical Pharmacology and Therapeutics

基　　金：浙江省自然科学基金资助项目(LQ16H310003);浙江省人民医院2015年优秀青年人员科研启动基金(zry2015B013);浙江省药学会医院药学专项科研资助项目(2016ZYY14);国家自然科学基金资助项目(81503521)

摘　　要：目的:构建S-亚硝基化谷胱甘肽(S-nitrosoglutathione,GSNO)诱导脑微血管内皮细胞的损伤模型,探讨丁苯酞(Butylphthalide,dL-NBP)对脑微血管内皮细胞的保护作用与可能的分子调控机制。方法:建立损伤bEnd.3细胞模型,以MTT法检测细胞存活率。DCFH染色检测细胞活性氧水平。Western blot法检测p-ERK,ERK,cleaved caspase 9,pro-caspase 9,cleaved caspase 3,pro-caspase 3蛋白表达。RT-PCR法检测糖皮质激素受体(GR)、SGK、MKP-1基因表达水平。结果:dL-NBP(5,10,20 μmol/L)可减少GSNO引起的脑微血管内皮细胞损伤,提高细胞存活率,减少ROS发生。此外,dL-NBP可激活SGK和MKP-1转录水平的增加,上调ERK磷酸化,抑制cleaved caspase 9,cleaved caspase 3蛋白上调。结论:dL-NBP对GSNO诱导的脑微血管内皮细胞损伤具有保护作用,其作用机制与激活PR活性下游基因SGK和MKP-1,上调ERK磷酸化水平,抑制caspase级联反应密切相关。AIM : To investigate the protection of Butylphthalide (dL-NBP) on oxidative damage in brain microvascular endothelial cells,and to further find the potential mechanisms of dL-NBP effects. METHODS : The brain microvascular endothelial cells were injured by GSNO.The cell viability was measured by MTT assay.ROS was observed by DCFH staining.ERK,p-ERK,cleaved caspase 9,pro-caspase 9,cleaved caspase 3 and pro-caspase 3 were detected by Western blot.The gene expressions of GR,SGK and MKP-1 were detected by RT-PCR assay. RESULTS :dL-NBP (10,20 μmol/L) protected endothelial cells against GSNO-induced injury and ROS release.dL-NBP increased SGK and MKP-1 gene expression.dL-NBP up-regulated ERK phosphorylation.dL-NBP inhibited cleaved caspase 9 and cleaved caspase 3. CONCLUSION : dL-NBP attunates bEnd.3 cells GSNO-induced injury,and its mechanism is related with activating GR relative SGK and MKP-1.dL-NBP increases ERK phosphorylation and down-regulates caspase decades.

关 键 词：丁苯酞 脑微血管内皮细胞 S-亚硝基化谷胱甘肽 ERK磷酸化 活性氧 

分 类 号：R743[医药卫生—神经病学与精神病学] R966[医药卫生—临床医学]