WU HAITAO, JIANG YONG, ZHANG XIAODONG, et al. APOE polymorphisms affects NF-κB expression in cultured mouse astrocytes in the early stage postinjury. [J]. Journal of army medical university, 2010, 32(2): 103-106.
WU HAITAO, JIANG YONG, ZHANG XIAODONG, et al. APOE polymorphisms affects NF-κB expression in cultured mouse astrocytes in the early stage postinjury. [J]. Journal of army medical university, 2010, 32(2): 103-106.DOI:
Objective To investigate the correlation between the polymorphisms of apolipoprotein E gene (APOE) and the expression of NF-κB of astrocytes in the early stage postinjury. Methods The CDS region of 3 APOE alleles was obtained by reverse transcription polymerase chain reaction (RT-PCR)
then the recombinant plasmid pEGFP-N1-APOE was constructed and identified by sequencing. Astrocytes were separated from APOE gene-knockout mice
and identified by immunocy
tochemistry method. The recombinant plasmids containing the 3 APOE alleles (APOEε2
APOEε3 and APOEε4) were respectively transfected into the isolated astrocytes with Lipofectamine 2000. Injury was produced by scratch injury to a monolayer of confluent astrocytes. RT-PCR was used to detect the expression of NF-κB mRNA in 12
24 and 48 h postinjury. Western blot analysis was used to detect the expression of NF-κB protein as well. Results The expression of NF-κB was observed at the 12th hour after injury
but there was no statistical difference among the 3 groups (
P
<
0.05). The expression was increased at the 24th hour postinjury. And compared with the other 2 groups
APOEε4 transfected astrocytes expressed higher level of NF-κB (
P
<
0.05). At the 48th hour after injury
the expression of NF-κB was increased further
and statistical difference was seen between APOEε4 and the other 2 groups (
P
<
0.05). Conclusion In the early stage postinjury
APOEε4 transfected astrocytes have higher expression of NF-κB at mRNA and protein levels than the cells with APOEε2 and APOEε3 transfection
suggesting APOEε4 carriers may have more severe deterioration and poorer prognosis due to early inflammatory response mediated by APOEε4.
关键词
载脂蛋白E基因多态性星形胶质细胞核因子kappa B
Keywords
apolipoprotein Epolymorphismastrocytesnuclear factor-kappa B