促红细胞生成素预处理对离体血管内皮细胞缺血再灌注模型中内皮型一氧化氮合酶表达的影响  

Erythropoietin preconditioning effecton the expression of endothelial nitric oxide synthase of the vascular endothelial cells in vitro with ischemia-reperfusion injury

作　　者：王源江 李红戈[2] 赵振强[1] 王埮[1] 李嫚 陈志斌[1] WANG Yuanjiang;LI Hongge;HAO Zhenqiang;WANG Tan;LI Man;CHEN Zhibin(Department of Neurology,the Affiliated Hospital of Hainan Medical College,Haikou 570102,China)

机构地区：[1]海南医学院第一附属医院神经内科,海口570102 [2]华中科技大学同济医学院附属协和医院神经内科,武汉430022

出　　处：《实用医学杂志》2018年第19期3163-3167,3171,共6页The Journal of Practical Medicine

基　　金：国家自然科学基金资助项目(编号:31260231);海南省科技合作专项资金项目(编号:KJHZ2015-09);海南医学院第一附属医院科研培育基金项目(编号:HYFYPY201505)

摘　　要：目的研究重组人促红细胞生成素预处理对离体血管内皮细胞缺血再灌注模型中内皮型一氧化氮合酶表达的影响及其可能的作用机制。方法体外培养人脐静脉内皮细胞株(Ea.hy926),采用MTT法检测细胞活性,细胞免疫荧光激光共聚焦荧光显微镜定位检测及Western Blot检测eNOS和Caspase-3在VECs中的表达。结果缺血再灌注损伤可明显降低VECs的细胞活力,而给予重组人促红细胞生成素预处理可显著改善缺血再灌注损伤下VECs的细胞活力;缺血再灌注损伤可显著减少VECs内eNOS表达、显著增加Caspase-3表达;而rhEPO预处理可显著提高缺血再灌注损伤下VECs内eNOS表达、显著减少Caspase-3表达。结论 rhEPO预处理可提高缺血再灌注损伤下内皮细胞的细胞活力,并能显著提高e NOS表达水平,提高血管内皮细胞对缺血再灌注损伤的耐受力。Objective To investigate recombinant human erythropoietin(rhEPO)preconditioning effect on the expression of endothelial nitric oxide synthase(eNOS)in the vascular endothelial cells(VECs)ischemia-reperfusion injury(I/R)model in vitro,and to clarify the possible mechanism.Methods The human umbilical veins endothelial cell line(Ea.hy926)was cultured in vitro.The cells survival rate was evaluated by MTT colorime-try.The expression of endothelial nitric oxide synthase(eNOS)and cysteine aspartic acic specific protease 3(Cas-pase-3)were detected by cell immunofluorescence and Western Blot.Results Compared with the normal control group,the cell viability of I/R group was significantly decreased.The cell viabilities of different concentrations rhE-PO-preconditioning groups increased significantly,compared with I/R group.Compared with the normal control group,the expression of eNOS on I/R group decreased significantly,but the expression of caspase-3 increased sig-nificantly.Compared with the I/R group,the expression of eNOS on different concentrations rhEPO preconditioning groups increased significantly,but the expression of caspase-3 decreased significantly.Conclusion rhEPO improves the cell viability of VECs and increases the expression of eNOS.It might be concluded that rhEPO can improve the tolerance of VECs for ischemia-reperfusion injury.

关 键 词：重组人促红细胞生成素 预处理 人脐静脉内皮细胞 缺血再灌注 内皮型一氧化氮合酶 缺血耐受力 

分 类 号：R743.3[医药卫生—神经病学与精神病学]