血管内皮生长因子基因转染骨髓间充质干细胞对慢性肾衰竭大鼠肾脏的修复作用

Repairing effect of bone marrow mesenehymal stem cells transfected with VEGF gene in rats with chronic renal failure

目的探讨血管内皮生长因子（vascularendothelialgrowthfactor，VEGF）基因转染骨髓间充质干细胞（mesenchymalstemcelis，MSC）对慢性肾衰竭（chronicrenalfailure，CRF）大鼠肾脏的修复作用。方法体外分离、培养大鼠MSC，Ad—VEGF转染MSC。SD大鼠随机分为假手术组（对照组）、慢性肾衰竭模型组（肾衰组）、慢性肾衰竭大鼠MSC移植组（MsC组）、慢性。肾衰竭竭大鼠AdVEGF注射组（VEGF组）和慢性肾衰竭大鼠Ad—VEGF转染的MSC移植组（转染组）。采用分阶段5／6肾切除术制备大鼠CRF动物模型。对照组与肾衰组于切除右肾之前从该侧肾动脉注射不含血清的DMEM培养液，其他3组分别给予MSC、Ad—VEGF和VEGF基因转染的MSC。8周后检测各组大鼠血肌酐（SCr）、血尿素氮（BUN）和24h尿蛋白，观察各组大鼠残肾组织病理形态，并用免疫印迹和RT—PCR方法检测各组大鼠残肾组织VEGFmRNA和蛋白的表达。结果MSC组和转染组SCr、BUN和尿蛋白均较。肾衰组降低（P〈0．05）；与MSC组比较，转染组SCr、BUN和尿蛋白降低更明显（P〈0．05）。肾衰组残肾组织VEGFmRNA和蛋白的表达较对照组显著减少（P〈0．05），MSC组和转染组残肾组织VEGFmRNA和蛋白的表达明显增加（与肾衰组比较，均P〈0．05），转染组残肾组织VEGFmRNA和蛋白的表达增加更明显（与M9C组比较，P〈0．05）。结论VEGF基因转染的MSC移植对CRF大鼠的肾脏有修复作用，这可能与VEGF在肾组织中高表达有关。

Objective To investigate the effects of mesenchymal stem cells （MSCs） transfected with vascular endothelial growth factor（VEGF） gone on repair of damaged kidney in rats with chronic renal failure（CRF）. Methyls MSCs isolated from rat bone marrow were cultured and transfeeted with the recombinant Ad5-hVEGF-EGFP. SD rats were randomly divided into five groups： sham operation group, CRF group, CRF ＋ MSCs group, Ad VEGF group, and MSCs ＋ VEGF group. The model of CRF was established by a two stage 5/6 nephrectomy procedure in rats. After 8 weeks, serum creatinine （SCr）, blood urea nitrogen（BUN） and 24-h urine protein were measured, pathological morphology of the residual kidney was observed, and VEGF expression in the residual kidney was detected by West ern blotting and reverse transcription-polymerase chain raction （RT-PCR）. Results SCr,BUN and 24- h urine protein levels in CRF ＋ MSCs group and MSCs ＋ VEGF group were significantly lower than those in CRF group （P〈0. 05）. SCr, BUN and 24-h urine protein levels in MSCs ＋ VEGF group were significantly lower than those in MSCs group （P〈0. 05）. As compared with sham operation group, the expression of VEGF mRNA and protein in the residual kidney was significantly decreased in CRF group （P〈0. 05）. As compared with CRF group, the expression of VEGF mRNA and protein in the residual kidney was significantly increased in CRF ＋ MSCs group and MSCs ＋ VEGF group （all P〈 0. 05）. As compared with CRF ＋ MSCs group, the expression of VEGF mRNA and protein in the resid- ual kidney was increased in MSCs ＋ VEGF group （P〈0. 05）. Conclusions Damaged kidney could be repaired by transplantation of MSCs transfected with VEGF gene in CRF rats probably by up^regula ting the VEGF expression in the residual kidney of CRF rats.