缺氧缺血对O-2A祖细胞膜铁转运蛋白1表达的影响  

Effect of hypoxia-ischemia on the expression of ferroportin 1 in O-2A progenitor cells

作　　者：林清[1] 张更[1] 邱荣晖[1] 王玮[1] 

机构地区：[1]福建医科大学基础医学院人体解剖学与组织学胚胎学系,神经生物学研究中心,福州350108

出　　处：《解剖学报》2013年第3期324-329,共6页Acta Anatomica Sinica

基　　金：福建医科大学苗圃科研基金资助项目(2010MP028);省属高校专项科研基金资助项目(jk2009013)

摘　　要：目的观察膜铁转运蛋白1(FPN1)在O-2A祖细胞缺氧缺血损伤后的表达变化,探讨其在O-2A祖细胞缺氧缺血损伤中的作用。方法体外培养O-2A祖细胞,以特异性抗体A2B5鉴定,观察FPN1在O-2A祖细胞上的定位表达;以糖氧剥夺(OGD)法建立缺氧缺血细胞模型,CCK8法检测细胞存活率;应用免疫荧光染色法、实时荧光定量PCR法和Western blotting法观察FPN1在细胞缺氧缺血后的表达变化。结果 FPN1定位表达于O-2A祖细胞的细胞膜、细胞质和突起;OGD3h、6h、12h及24h细胞存活率呈时间依赖性降低(P<0.05);OGD12h内细胞FPN1免疫荧光强度进行性减弱;与OGD0h相比,FPN1 mRNA和蛋白水平在OGD3h表达下调,OGD6h进一步降低,OGD12h最低,差异具有统计学意义(P<0.05)。结论 O-2A祖细胞缺氧缺血损伤后FPN1表达明显下调,细胞存活率显著降低,提示FPN1可能参与O-2A祖细胞的缺氧缺血损伤过程。Objective To investigate the FPN1 expression and its role in O-2A progenitor cells after hypoxic- ischemic injury. Methods O-2A progenitor cells were cultured in vitro, indentified with A2B5 antibody and investigated by the localization of FPN1. Hypoxic-ischemic cell models were established by using the oxygen-glucose deprivation （OGD） method and the cell viability was assessed by the CCK-8 method. The expression of FPN1 in O-2A progenitor cells after hypoxia-ischemia was detected by immunofiuorescent staining, quantitative real-time polymerase chain reaction analysis and Western blotting analysis. Results FPN1 was localized at the cell membrane, and in the cytoplasm and processes of O-2A progenitor cells. The cell viability decreased with time-dependence after 3hours, 6hours, 12hours and 24hours of OGD （ P 〈 0.05）. The FPN1 immunofluorescence intensity of O-2A progenitor cells decreased progressively within 12hours of OGD. The FPN1 mRNA and protein levels downregulated with time-dependence after 3hours, 6hours and 12hours of OGD （ P 〈 0.05 ）. Conclusion The level of FPN1 expression is down-regulated and cell viability decreased significantly with time-dependence after OGD, which suggests that FPN1 may play a role in the hypoxic-ischemic injury of O-2A progenitor ceils.

关 键 词：膜铁转运蛋白1 缺氧缺血 实时定量一聚合酶链反应 免疫印迹法 O-2A祖细胞 

分 类 号：R722.6[医药卫生—儿科]