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[Molecular bypass, the application of VEGF to gene therapy of limb ischemia]
Objective:
To investigate the feasibility of using VEGF gene directly for treating limb ischemia.
Methods:
The human VEGF165 cDNA was cloned into the eukaryotic expression vector pcDNA3 under the control of cytomegalovirus immediate-early promoter/enhancer. When gene suture was used, the recombinant plasmid was transferred into the hindlimb's adductor of rat, of which the distal extreme of external iliac artery was ligated and the femoral artery was completely excised. With RT-PCR, Western blot, immunohistochemistry analysis, and angiography, the expression and biological effects of VEGF gene in experimental animals were investigated.
Results:
Seven days after initiation of therapy, the transcription and expression of VEGF gene in experimental group were significantly higher than those of control groups. The results from angiography, feet temperature measuring and observation of clinical symptom showed that the transfer of VEGF gene can stimulate the formation of focal neovessles, establish colleteral circulation, augment blood perfusion, and ameliorate distal limb necrosis.
Conclusion:
These findings reveal that the "molecular bypass" using VEGF gene may represent a potential therapeutic modality for treating tissue ischemia.