Effect of human hypoxia inducible factor-1? gene transfection on apoptosis of A?25-35 induced hippocampal neurons of primary culture
- VernacularTitle:HIF-1?基因转染对?-淀粉样蛋白25-35诱导原代培养海马神经细胞凋亡的影响
- Author:
Lingyun LIU
;
Xiqing CHAI
;
Qingjun LI
;
Guihua CHENG
- Publication Type:Journal Article
- Keywords:
hypoxia inducible factor-1?;
primary culture;
hippocampus;
transfection
- From:Journal of Third Military Medical University
2003;0(07):-
- CountryChina
- Language:Chinese
-
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
