Ye Lei, Haider Husnain Kh, Tan RuSan, Toh WeeChi, Law Peter K, Tan WeeBeng, Su LiPing, Zhang Wei, Ge RuoWen, Zhang Yong, Lim YeanTeng, Sim Eugene K W
National University Medical Institutes, National University of Singapore, Singapore.
Circulation. 2007 Sep 11;116(11 Suppl):I113-20. doi: 10.1161/CIRCULATIONAHA.106.680124.
We investigated the feasibility and efficacy of polyethylenimine (PEI) based human vascular endothelial growth factor-165 (hVEGF165) gene transfer into human skeletal myoblasts (HSM) for cell based delivery to the infarcted myocardium.
Based on optimized transfection procedure using enhanced green fluorescent protein (pEGFP), HSM were transfected with plasmid-hVEGF165 (phVEGF165) carried by PEI (PEI-phVEGF165) nanoparticles. The transfected HSM were characterized for transfection and expression of hVEGF165 in vitro and transplanted into rat heart model of acute myocardial infarction (AMI): group-1=DMEM injection, group-2= HSM transplantation, group-3= PEI-phVEGF165-transfected HSM (PEI-phVEGF165 myoblast) transplantation. A total of 48 rats received cyclosporine injection from 3 days before and until 4 weeks after cell transplantation. Echocardiography was performed to assess the heart function. Animals were sacrificed for molecular and histological studies on the heart tissue at 4 weeks after treatment. Based on optimized transfection conditions, transfected HSM expressed hVEGF165 for 18 days with >90% cell viability in vitro. Apoptotic index was reduced in group-2 and group-3 as compared with group-1. Blood vessel density (x400) by immunostaining for PECAM-1 in group-3 was significantly higher (P=0.043 for both) as compared with group-1 and group-2 at 4 weeks. Regional blood flow (ml/min/g) in the left ventricular anterior wall was higher in group-3 (P=0.043 for both) as compared with group-1 and group-2. Improved ejection fraction was achieved in group-3 (58.44+/-4.92%) as compared with group-1 (P=0.004).
PEI nanoparticle mediated hVEGF165 gene transfer into HSM is feasible and safe. It may serve as a novel and efficient alternative for angiomyogenesis in cardiac repair.
我们研究了基于聚乙烯亚胺（PEI）的人血管内皮生长因子-165（hVEGF165）基因转导入人骨骼肌成肌细胞（HSM），用于以细胞为基础向梗死心肌递送的可行性和疗效。
基于使用增强型绿色荧光蛋白（pEGFP）的优化转染程序，用PEI（PEI-phVEGF165）纳米颗粒携带的质粒-hVEGF165（phVEGF165）转染HSM。对转染的HSM进行体外hVEGF165转染和表达的表征，并将其移植到急性心肌梗死（AMI）大鼠心脏模型中：第1组=注射DMEM，第2组=移植HSM，第3组=移植PEI-phVEGF165转染的HSM（PEI-phVEGF165成肌细胞）。总共48只大鼠在细胞移植前3天直至移植后4周接受环孢素注射。进行超声心动图以评估心脏功能。在治疗后4周处死动物，对心脏组织进行分子和组织学研究。基于优化的转染条件，转染的HSM在体外表达hVEGF165达18天，细胞活力>90%。与第1组相比，第2组和第3组的凋亡指数降低。在4周时，第3组通过PECAM-1免疫染色的血管密度（×400）显著高于第1组和第2组（两者P=0.043）。第3组左心室前壁的局部血流量（ml/min/g）高于第1组和第2组（两者P=0.043）。与第1组相比，第3组的射血分数提高（58.44±4.92%）（P=0.004）。
PEI纳米颗粒介导的hVEGF165基因转导入HSM是可行且安全的。它可能作为心脏修复中血管生成的一种新型高效替代方法。
