Xin-jie Tan1, Chang-lin Hu. 1. Department of Neurology, Second Affiliated Hospital, Chongqing University of Medical Sciences, Chongqing 400010, China.
Abstract
OBJECTIVE: To construct a recombinant adenoviral vector carrying HIF-1alpha gene and explore the therapeutic effect of HIF-1alpha on focal cerebral ischemia in adult rats. METHODS: The AdEasy System was used to construct the recombinant adenoviral vector carrying HIF-1alpha gene and green fluorescent protein and PCR was used to identify the HIF-1alpha gene. Middle cerebral artery occlusion (MCAo) and reperfusion models were established and divided into Ad-HIF-1alpha group, Ad group and NS group. After Ad-HIF-1alpha, Ad and NS were injected into the ischemic ventricle, expression of Ad-HIF-1alpha was observed and its therapeutic effect was evaluated by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining and neurological severity scores. RESULTS: GFP expression distributed apart from the ventricle and reached a peak at 14 days and persisted for about 4 weeks under fluorescent microscope. The neurological severity scores was 2.4 +/- 0.5 at 24 h in Ad-HIF-1alpha group and there was no statistical significance compared with Ad group (2.6 +/- 0.5) and NS group (2.7 +/- 0.7) (P > 0.05). The scores were 1.6 +/- 0.7 at 48 h and 0.9 +/- 0.6 at 72 h in Ad-HIF-1alpha group, and there were statistical significance compared with Ad group (2.9 +/- 0.6 and 3.2 +/- 0.6 respectively) and NS group (3.0 +/- 0.7 and 3.2 +/- 0.8) (P < 0.05). The infarct volume was 81.2 mm(3) +/- 1.4 mm(3) at 72 h in Ad-HIF-1alpha group and there was statistical significance compared with Ad group (173.9 mm(3) +/- 1.3 mm(3)) and NS group (171.7 mm(3) +/- 6.2 mm(3)) (P < 0.05). CONCLUSION: HIF-1alpha gene had definite therapeutic effect on focal cerebral ischemia in adult rats, which settles a foundation for next HIF-1alpha gene study and clinic application.
OBJECTIVE: To construct a recombinant adenoviral vector carrying HIF-1alpha gene and explore the therapeutic effect of HIF-1alpha on focal cerebral ischemia in adult rats. METHODS: The AdEasy System was used to construct the recombinant adenoviral vector carrying HIF-1alpha gene and green fluorescent protein and PCR was used to identify the HIF-1alpha gene. Middle cerebral artery occlusion (MCAo) and reperfusion models were established and divided into Ad-HIF-1alpha group, Ad group and NS group. After Ad-HIF-1alpha, Ad and NS were injected into the ischemic ventricle, expression of Ad-HIF-1alpha was observed and its therapeutic effect was evaluated by 2, 3, 5-triphenyltetrazolium chloride (TTC) staining and neurological severity scores. RESULTS: GFP expression distributed apart from the ventricle and reached a peak at 14 days and persisted for about 4 weeks under fluorescent microscope. The neurological severity scores was 2.4 +/- 0.5 at 24 h in Ad-HIF-1alpha group and there was no statistical significance compared with Ad group (2.6 +/- 0.5) and NS group (2.7 +/- 0.7) (P > 0.05). The scores were 1.6 +/- 0.7 at 48 h and 0.9 +/- 0.6 at 72 h in Ad-HIF-1alpha group, and there were statistical significance compared with Ad group (2.9 +/- 0.6 and 3.2 +/- 0.6 respectively) and NS group (3.0 +/- 0.7 and 3.2 +/- 0.8) (P < 0.05). The infarct volume was 81.2 mm(3) +/- 1.4 mm(3) at 72 h in Ad-HIF-1alpha group and there was statistical significance compared with Ad group (173.9 mm(3) +/- 1.3 mm(3)) and NS group (171.7 mm(3) +/- 6.2 mm(3)) (P < 0.05). CONCLUSION:HIF-1alpha gene had definite therapeutic effect on focal cerebral ischemia in adult rats, which settles a foundation for next HIF-1alpha gene study and clinic application.