LIU Ling-yun,CHAI Xi-qing,LI Qing-jun,CHENG Gui-hua.Effect of human hypoxia inducible factor-1α gene transfection on apoptosis of Aβ25-35 induced hippocampal neurons of primary culture[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(07):595-598.
LIU Ling-yun,CHAI Xi-qing,LI Qing-jun,CHENG Gui-hua.Effect of human hypoxia inducible factor-1α gene transfection on apoptosis of Aβ25-35 induced hippocampal neurons of primary culture[J].J Amry Med Univ (J Third Mil Med Univ),2007,29(07):595-598.DOI:
Objective To construct the eukaryotic expression vector for human hypoxia inducible factor-1α gene (pSNAV-HIF-1α)
and to investigate the apoptosis and intracellular calcium concentration of the transfected Aβ25-35 induced hippocampal neurons of primary culture. Methods Human hypoxia inducible factor-1α gene from pBSKhHIF1αT7 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 Aβ25-35 for treatment. Empty pSNAV2.0 vector was treated the same way as pSNAV-HIF-1α as a control. The expression level of HIF-1α 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-1α was successfully constructed by restriction enzyme digestion
PCR and DNA sequencing. The expression level of HIF-1α protein was significantly increased in transfected hippocampal neurons of primary culture (P0.05). The up-regulation of apoptosis and intracellular calcium concentration induced by Aβ25-35 were significantly decreased in transfected hippocampal neurons of primary culture (P0.05). Conclusion Human hypoxia inducible factor-1α gene transfection could inhibit Aβ25-35 induced apoptosis of hippocampal neurons of primary culture by suppressing the up-regulation of intracellular calcium concentration.