Expression of hVEGF and its protein in transfected ischemic muscle by phVEGF_(165) / 中国现代普通外科进展

ABSTRACT

Objective:To investigate a valuable method of gene therapy to treat chronic arterial ischemic diseases in lower extremity.Methods:VEGF was selected as the target therapeutic gene.Recombinant plasmid phVEGF 165 constructed in vitro as the eukaryotic vector of human VEGF cDNA,was directly injected into the ischemic hindlimb muscles in rabbit models.In situ hybridization and RT PCR were carried out to detect the expression of hVEGF 165 in the muscle.Immunohistochemistry assay and Western blotting were carried out to detect the expression of hVEGF 165 protein.Results:The expression of external hVEGF 165 peaked at l week and sustained 2～3 weeks.The expression of protein peaked at 2 weeks and lasted 4 weeks after injection.Conclusion:Skeletal muscle can extract and express external hVEGF 165 ,and further translate the external gene into protein.The effect of direct intramuscular injection of phVEGF 165 is limited in local muscles,and no longer than 1 month.So this method of gene therapy is quite safer.