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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 feerroportin 1 in O-2A progenitor cells
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
immunofluorescent 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 cells.
膜铁转运蛋白1 / 缺氧缺血 / 实时定量-聚合酶链反应 / 免疫印迹法 / O-2A祖细胞
Ferroportin 1 / Hypoxia-ischemia / Real-time PCR / Western blotting / Oligodendrocyte-type-2 astrocyte progenitor cell
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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）；福建省卫生厅青年科研基金资助项目（2009-1-28）
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