摘要
Abstract
Objective To investigate the mechanism and regulation of hTERT modified endothelial progeni-tor cells (EPCs) transplantation on ischemic limb of SD rats. Methods EPCs were cultured and identified af-ter Lipofectamine stably transfected PLXSN-hTERT EPCs expression by Western blot identification of exoge-nous human hTERT gene in endothelial progenitor cells. Sixty-four male SD rats were randomly divided into 4 groups: A, B, C and D. Each group had a total of 16 rats. After ligation of the femoral artery and branches of the left hind limbs, the hind limb ischemia model was constructed, the adductor and gastrocnemius muscle were in-jected 5 points, each point injection 60 L. A group rats were injected with 300 L medium, B group rats were in-jected with 5 g of PLXSN-hTERT gene DNA- liposome complexes, C group rats were injected with 4 × 106 EPCs, and D group rats were injected with 4 × 106 hTERT gene EPCs. RT-PCR angiography was performed to detect the expression of hTERT gene and neovascularization in 4 weeks after transplantation, and the survival of PKH-26 labeled EPCs was evaluated by fluorescence microscope. Results All of the 64 rats were successful-ly constructed, and the results of Western blot showed that human endothelial progenitor cells transfected with human hTERT gene could express hTERT in vitro 4 weeks after transplantation and angiography. A group rat femoral artery and its branches were not visualized, B group and C group showed moderate neovascularization (ef-fects of the two groups were similar), D group can further enhance the effect. The number of PKH-26 positive cells were: [A,B,C and D four groups (0 ± 0.00),(0 ± 0.00),(27.46 ± 5.48),(54.37 ± 8.34)/HP, P<0.05]. In vivo ex-pression of hTERT gene in RT-PCR, D group and B group were transfected with hTERT gene amplification, hTERT mRNA relative content of D group was higher in B group (0.89 ± 0.06, 0.29 ± 0.04, P<0.05). Con-clusion Using Htert modified EPCs transplantation to treat limb ischemia in rats, can make hTERT gene stable and effective expression, and its effect is better than the direct gene injection.关键词
内皮祖细胞/移植/缺血/端粒酶逆转录酶/大鼠Key words
Endothelial progenitor cells/transplantation/ischemia/telomerase reverse transcriptase/rat分类
生物科学
