解剖学报 ›› 2013, Vol. 44 ›› Issue (3 ): 324-329.doi: 10.3969/j.issn.0529-1356.2013.03.006

• 神经生物学 • 上一篇    下一篇

缺氧缺血对O-2A祖细胞膜铁转运蛋白1表达的影响

林清 张更 邱荣晖 王玮*   

  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
  • 收稿日期:2012-08-13 修回日期:2012-09-18 出版日期:2013-06-06 发布日期:2013-07-16
  • 通讯作者: 林清 E-mail:linqing522@126.com
  • 基金资助:

    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)

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

LIN Qing  ZHANG Geng  QIU Rong-hui  WANG Wei*   

  1. Department of Human Anatomy, Histology and Embryology, School of Preclinical Medicine,Research Center for Neurobiology, Fujian Medical University, Fuzhou 350108, China
  • Received:2012-08-13 Revised:2012-09-18 Online:2013-06-06 Published:2013-07-16

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。

关键词: 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

Abstract:

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.

Key words: Ferroportin 1, Hypoxia-ischemia, Real-time PCR, Western blotting, Oligodendrocyte-type-2 astrocyte progenitor cell