Effect of human hypoxia inducible factor-1? gene transfection on apoptosis of A?25-35 induced hippocampal neurons of primary culture / 第三军医大学学报
Journal of Third Military Medical University
;
(24)2003.
Artigo
em Chinês
| WPRIM
| ID: wpr-561283
ABSTRACT
Objective To construct the eukaryotic expression vector for human hypoxia inducible factor-1? gene (pSNAV-HIF-1?), and to investigate the apoptosis and intracellular calcium concentration of the transfected A?25-35 induced hippocampal neurons of primary culture. Methods Human hypoxia inducible factor-1? gene from pBSKhHIF1?T7 was inserted into pSNAV2.0 by the method of gene recombination, then the constructed vector was transfected into hippocampal neurons of primary culture and followed by A?25-35 for treatment. Empty pSNAV2.0 vector was treated the same way as pSNAV-HIF-1? as a control. The expression level of HIF-1? protein was assayed by Western blotting. Apoptosis was detected by flow cytometry. Intracellular calcium concentration was determined by laser scanning confocal microscopy with Fluo-3/AM as the fluorescent dye. Results It was shown that pSNAV-HIF-1? was successfully constructed by restriction enzyme digestion, PCR and DNA sequencing. The expression level of HIF-1? protein was significantly increased in transfected hippocampal neurons of primary culture (P
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Índice:
WPRIM (Pacífico Ocidental)
Idioma:
Chinês
Revista:
Journal of Third Military Medical University
Ano de publicação:
2003
Tipo de documento:
Artigo
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