Triple Point-Mutants of Hypoxia-Inducible Factor-1Accelerate In Vivo Angiogenesis in Bone Defect Regions

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• 作者：Chen Li (1)
  Danping Liu (2)
  Zheng Zhang (2)
  Guoxian Wang (3)
  Na Xu (4)
  
• 关键词：Hypoxia ; inducible factor 1 Mutagenesis ; Recombinant adenovirus vector ; Bone marrow mesenchymal stem cells ; Angiogenesis
• 刊名：Cell Biochemistry and Biophysics
• 出版年：2013
• 出版时间：November 2013
• 年：2013
• 卷：67
• 期：2
• 页码：557-566
• 全文大小：
• 作者单位：Chen Li (1)
  Danping Liu (2)
  Zheng Zhang (2)
  Guoxian Wang (3)
  Na Xu (4)
  
  1. Central Laboratory, The First Affiliated Hospital of Liaoning Medical College, Jinzhou, China
  2. Orthopedic Department, The First Affiliated Hospital of Liaoning Medical College, No. 2, Renming Street Wuduan, Guta District, Jinzhou, 121001, China
  3. Pharmacology Department, The Basic Medicine College of Liaoning Medical College, Jinzhou, China
  4. Radiology Department, The First Affiliated Hospital of Liaoning Medical College, Jinzhou, China
  
• ISSN：1559-0283

文摘

To investigate the functions of triple point-mutants of hypoxia-inducible factor 1(HIF1 in angiogenesis in bone defect regions under normoxic conditions.1. Triple point-mutations (in amino acids 402, 564, and 803) in the HIF1coding sequence (CDS) were induced by polymerase chain reaction. The triple mutant HIF1(402/564/803) was inserted into the adenovirus pAdEasy-1 system for complete viral packaging and titer measurements. 2. For the in vitro experiment, rabbit bone marrow mesenchymal stem cells (MSCs) were divided into four experimental groups. The efficiency of infection was observed by the expression of human renilla reniformis green fluorescent protein (hrGFP). The HIF1mRNA, protein and VEGF protein expression levels in infected cells in each experimental group were measured. 3. As in the in vivo experiment, the MSCs were divided into four groups and infected with the viral solutions from each complementary in vitro group and cultured under normoxic conditions. The MSCs were used as seed cells and transplanted into an apatiteollastonite magnetic bioactive glasseramic (AW MGC) vector to construct artificial tissue-engineering scaffolds that were then implanted into the in vivo rabbit radial bone defect model. The animals from each group were killed 8weeks after the surgery, and the tissues from the implantation region were harvested for the evaluation of the angiogenesis. 1. The 402,564, and 803 amino acids in CDS area were point mutated into alanine; three types of recombinant adenovirus were successfully constructed, packaged, and characterized. 2. The expression levels of HIF1mRNA in A and B groups were significantly higher than those in the C and D groups (P<0.05). The HIF1and VEGF protein expression levels in A group were significantly higher than those in the other three groups (P<0.05). 3. There was prominent angiogenesis in bone defect regions in group A animals. 1. Triple point-mutants of HIF1efficiently expressed functional proteins under normoxic conditions.2. Triple point-mutants HIF1effectively promoted in vivo angiogenesis in bone defect regions.