Objective:To investigate the regulatory effects and possible mechanisms of recombinant adenovirus-mediated hypoxia inducible factor-1 alpha(HIF-1α)transfection on HIF-1α in post-hypoxic rat cortical neurons.Methods:(1)Preparation of rats' cerebral cortical neurons in primary culture and hypoxia model.Recombinant adenovirus-mediated HIF-1α(AdHIF-1α)transfection and recombinant Ad transduction in normal and hypoxic cells.Then they will be divided into normal control group, hypoxia group, the recombinant adenovirus hypoxia-inducible factor-1α(AdHIF-1α)gene transfection group and recombinant adenovirus empty vector(Ad)transfection group.(2)To observe the transfection of AdHIF-1α/ Ad under fluorescence microscopy.The expression of hypoxia-inducible factor-1 alpha(HIF-1α)were observed by Western blot analysis in each group of neurons at 12 h、24 h、48 h and 72 h.Results:Hypoxic neurons transfected with Ad/AdHIF-lα showed the obvious expression 48 h under fluorescence microscopy.At each time of the AdHIF-1α gene transfection group, the expression of HIF-1α is significantly increased, the positive expression emerges at 12 h, the expression increased at 24 h, and it reaches the peak at 48 h, until 72 h declines, and it has statistical significance compared with the hypoxia group( P<0.01, P<0.05), but the Ad group has no statistically significant differences compared with the hypoxia group. Conclusions:The transfection of recombinant adenovirus-mediated HIF-1α gene can significantly increase the expression levels of HIF-1α in hypoxic neurons, and it may play a protective role in the neurons after hypoxia.

Objective:To investigate the transfection of recombinant adenovirus vectors containing the hypoxia inducible factor-1α gene (AdHIF-1α)in rat brain microvascular endothelial cells(BMECs) and to provide a theoretical basis for the treatment of hypoxic BMECs by AdHIF-1α.Methods:Rat BMECs were isolated, identified, and cultured in a maintenance medium containing 100 μmol/L cobalt dichloride (CoCl 2), establishing a hypoxia model of BMECs; then AdHIF-1α was transfected into hypoxic BMECs.The transfection of fluorescent protein was observed under a fluorescence microscope. Results:Transfection of AdHIF-1α into BMECs was monitored under a fluorescence microscope at 12 h, 24 h, 48 h and 72 h, respectively.Minor fluorescence began to appear at 12 h (0.13±0.01), and the fluorescence expression increased at 24 h (0.46±0.03, q=25.88, P<0.01), was most obvious at 48 h (0.97±0.05, q=40.00, P<0.01), and decreased at 72 h (0.38±0.02, q=46.28, P<0.01). Conclusions:Recombinant adenovirus vectors containing AdHIF-1α can be transfected into hypoxic BMECs in vitro.