Abstract:

BACKGROUND: In vivo experiments of transgenic mice have shown that recombinant hypoxia-inducible factor 1 alpha (HIF-1α) can promote angiogenesis under the conditions of normal skin morphology and function.
OBJECTIVE: To construct a new adenovirus eukaryotic expression vector that can co-express HIF-1α mutant protein and human renilla reniformis green fluorescent protein (hrGFP) reporter molecule under normoxia conditions to transfect bone marrow mesenchymal stem cells (BMSCs) of the Sprague-Dawley rats. 
METHODS: A successful recombinant adenovirus eukaryotic expression vector pAd-HIF1αmu-IRES-hrGFP-1 was constructed and transfected into human embryonic kidney 293A cells (HEK293A) using Lipofectamine 2 000 for packaging the virus. The recombinant adenovirus eukaryotic expression vector was transfected into BMSCs of the rats at the best value multiplicity of infection 50. Three control groups were established: positive control group transfected with Ad-CMV-HIF1α-IRES-hrGFP-1, negative control group transfected with Ad-CMV-IRES-hrGFP-1 and blank control group without transfection.
RESULTS AND CONCLUSION: ①The adenovirus vectors were transfected into the HEK293A cells with packaging successfully and a large amount of green fluoresence expression was found in the cells. ②The protein expression in the cells transfected with mutant adenovirus HIF-1α expression vector under normoxia conditions was higher than that of the three control groups. The difference among the three control groups had no significance(P > 0.05). To transfect mutant adenovirus eukaryotic expression vector pAd-HIF1αmu-IRES-hrGFP-1 into the HEK293A cells with pakaging successfully can induce the great and efficient expression of HIF-1α gene under normoxia conditions.