Human mutant hypoxia inducible factor-1α in rat hindlimb ischemia model of angiogenesis in.docVIP

 PAGE \* MERGEFORMAT 7 Human mutant hypoxia inducible factor-1α in rat hindlimb ischemia model of angiogenesis in [Abstract] Objective: To explore the 564 and 402 double mutation of the HIF  1α gene (Ad  HIF  1α  564Ala  402Ala, referred to as Ad  H564/402) in rats with acute lower limb ischemia in the expression and vascular nascent. Methods: ① the pre-built Ad  H564/402 in HEK293A cells were amplified by using cesium chloride density gradient centrifugation for purification of adenovirus. using electron microscopy, PCR and sequencing methods to identify and use spectrophotometric Determination of total virus titer; ② Construction of acute lower limb ischemia model in rats were randomly divided into four groups, each with Ad  LacZ, Ad  H0, Ad  H564/402 and normal saline (NS) operated leg bones transfected muscle, in after transfection 1, 3, 5, 7 d, application of RT  PCR determination of skeletal muscle HIF  1α mRNA expression levels; ③ after gene transfection 28 d, selective arteriography and lower limb vascular casting of vascular density. Results: ① The amplified and purified gene mutation zone information is preserved, the final titer (6.29 ± 0.26) * 1014 OPU (optical particle unit, optical particle unit) / L. ② mutant Ad  H564 / The relative expression levels of 402 higher than the wild-type HIF  1α gene (P = 0.000); and the 7th day to the highest (P = 0.000); by any two different groups and different gene transfected to express the number of days between the volume has significant statistical difference (P = 0.000). ③ gene transfer 28 d after angiography and vascular casting results: Ad  H564/402 group of small blood vessels in the villi is greater than the number of Ad  H0 Group and other groups. Conclusion: ① exogenous mutant Ad  H564/402 gene can promote the in vivo HIF  1α mRNA expression. ② exogenous mutant Ad  H564/402 ischemic skeletal muscle gene can promote collateral blood vessel formation an