摘要
背景：骨髓间充质干细胞作为基因载体，有利于外源基因保持良好的遗传稳定性，如能将血管内皮细胞生长因子基因转入间充质干细胞内移植到心肌梗死区，可望获得基因和细胞治疗相互协同促进的作用。目的：观察转染血管内皮细胞生长因子基因的骨髓间充质干细胞移植大鼠心肌梗死区血管新生及血管内皮细胞生长因子基因的体内表达。设计、时间及地点：实验于2004／2007年在湖南省儿童医院儿科研究所，中南大学湘雅二医院中心实验室完成的随机对照实验。材料：实验动物为雄性清洁级Wistar近交系大鼠。以冠脉结扎法制备大鼠心肌梗死模型。方法：以密度梯度离心与贴壁培养法获得大鼠骨髓间充质干细胞。当间充质干细胞生长汇合率为80％～90％时以脂转法转染pcDNA3A-hVEGF165质粒。心肌梗死模型大鼠随机分4组进行干预：联合组于心肌梗死区移植转染血管内皮细胞生长因子基因的间充质干细胞，细胞组移植间充质干细胞、基因组注射脂质体一pcDNA31—VEGF165DNA复合物，对照组注射等容积培养液。主要观察指标：移植4周后，各组大鼠心肌梗死区毛细血管密度，血管内皮细胞生长因子基因表达。结果：大鼠心肌梗死区毛细血管密度以联合组和基因组最高，高于细胞组（P=0．001，0.029），细胞组高于对照组（P=0．028）。RT-PCR显示血管内皮细胞生长因子基因体内的表达从高到低依次为联合组、基因组、细胞组和对照组。结论：①作为血管内皮细胞生长因子基因的载体，骨髓间充质干细胞有利于其的稳定表达。②转染血管内皮细胞生长因子基因的骨髓间充质干细胞移植有利于大鼠心肌梗死区的血管新生，其疗效优于单独应用基因或细胞治疗。
BACKGROUND： Bone marrow mesenchymal stem cells （BMSCs） as a genetic cartier are beneficial tO keep a genetic stability of exogenous gene. BMSCs can transfer human vascular endothelial growth factor （hVEGF） gene into the myocardial infarcted area. It is possible to obtain a cooperation of gene and cell therapy.
OBJECTIVE： To investigate the angiogenesis at myocardial infarcted area of rats undergoing the transplantation of BMSCs transfected with hVEGF gene and in vivo expression of VEGF gene.
DESIGNs TIME AND SETTING： A randomized control experiment was performed at the Institute of Paediatrics of Hunan Children＇s Hospital and Central Laboratory of Xiangya Second Hospital of Central South University from 2004 to 2007.
MATERIALS： Male clean inbred strain Wistar rats were selected in the study. Rat models of myocardial infarction were established by ligation of coronary artery.
METHODS： BMSCs were harvested from rats by density gradient centrifugation and adherent culture. Plasmid pcDNA3.1-hVEGF165 were transfected at 80%-90% cell confluence. Rat models of myocardial infarction, constructed by ligation of the left anterior descending coronary artery, were randomly divided into 4 groups. 2 weeks after the ligation,BMSCs transfected with hVEGF gene were injected into rat models in the combination group at the myocardial infarction zone. BMSCs were injected into cell group. Liposome-pcDNA3.1-VEGF165 DNA compound was injected into gene group. Culture medium was injected into control group.
MAIN OUTCOME MEASURES： Capillary density and VEGF gene expression in the myocardial infarcted area of rats in each group four weeks after injection.
RESULTS： Capillary density was the highest at rat myocardial infarcted area in the combination and gene groups, followed by the cell group （P=0.001, 0.029） and the control group （P--0.028）. Reverse transcriptase-polymerase chain reaction （RT-PCR） showed that the hVEGF165 gene was expressed in the combination, gene, cell and control groups from higher level to lower level in order.
CONCLUSION： BMSCs as the vector of VEGF gene are beneficial to its stable expression. Transplantation of BMSCs transfected with hVEGF gene is helpful for angiogenesis at myocardial infarcted area of rats. Its outcome is better than gene or cell therapy alone.
出处
《中国组织工程研究与临床康复》
CAS
CSCD
北大核心
2008年第25期4979-4982,共4页
Journal of Clinical Rehabilitative Tissue Engineering Research
基金
湖南省科技基金项目(03TCY2012)课题名称:鼠骨髓间充质干细胞转染血管内皮细胞生长因子基因治疗缺血性心脏病的实验研究~~
