The Research on Treatment of Lentivirus-mediated hVEGF-165 Gene Transferring Mesenchymal Stem Cells on Rat Hind Limb Ischemia
|School||Zunyi Medical College,|
|Keywords||Rats Hindlimb ischemia Vascular endothelial growth factor gene -165 Mesenchymal stem cells|
Objective:Detect and analyze the therapeutic effect of bone marrow mesenchymal stem cells which carry human vascular endothelial growth factor-165 (hVEGF-165) gene on hind limb ischemia, and provide new ideas for the clinical treatment of lower limb’s ischemia.Method:The experiment set up three groups, which were group BMSC carrying hVEGF-165 gene, group BMSC and blank control group, and each group contains 8 samples for a total of 48 limbs. The ischemia model established by ligating the bifurcation above of the abdominal aorta of SD rats, detected and confirmed by postoperative observe, DSA angiography and measured femoral artery blood flow velocity and resistance index. With lentivirus-mediated hVEGF-165 gene transferring to infect rat bone marrow mesenchymal stem cells, detected the transferring rate of VEGF-165 gene under fluorescence microscope by fluorescent protein GFP; then, transplanted the stem cells into the ischemic limb by multi-point injection into rat hind limb muscles’method, and measured the effect by observing, DSA angiography, measured femoral artery blood flow velocity and resistance index one month later.Results:1, Fluorescence microscope detection displayed that the hVEGF-165 gene transferring BMSCs positive rate up to 85% and above;2, The ischemia model established by ligating the bifurcation above of the abdominal aorta, all the rats appeared limping, toe necrosis, and most appeared ischemic ulcers 30 days later. Color Doppler ultrasound, angiography and other tests displayed that after ligation surgery, blood flow velocity and resistance index of experimental rat’s hind limb significantly decreased, Vascular density was significantly reduced;3,30 days after the transplantation, efficacy test showed that the blood flow velocity.resistance index and vascular density of genetically modified BMSC group and BMSC group were significantly higher than the blank control group in rat’s hind limb, and genetically modified BMSC group higher than BMSC group (p<0.01).Conclusion:The positive rate of lentivirus-mediated hVEGF-165 gene transferring to infect BMSC is high; The ischemia model of ligating the bifurcation above of the abdominal aorta of SD rats had obvious ischemic effect, long duration; Under the same transplantation conditions, genetically modified BMSC group and BMSC group both can improve the blood supply of hindlimb ischemia, but BMSCs with hVEGF--165 gene owns advantage in helping vascular regeneration.