【摘要】 背景:骨髓间充质干细胞易于被外源基因转染,且具有向损伤组织趋化聚集的特性,便于将目的基因导入其中并发挥治疗性作用,可以通过分化为血管内皮细胞修复损伤的血管内膜。目的:探讨慢病毒载体介导的血管生长素1基因修饰骨髓间充质干细胞移植后,对大鼠颈动脉粥样硬化的影响。设计、时间及地点:随机对照动物实验,于2007-02/2008-05在福建省高血压研究所及福建医大附属第一医院中心实验室完成。材料:4周龄雄性SD大鼠32只,8只用于制备骨髓间充质干细胞,剩余24只随机分为3组:模型对照组、空载体组、血管生长素1转染组,8只/组。方法:以重组慢病毒为载体构建血管生长素1基因工程干细胞,以绿色荧光蛋白为报告基因,调整细胞密度为(1.0～2.0)×1010L-1。3组大鼠均采用球囊损伤法+高脂饮食构建颈动脉粥样硬化模型,造模后即刻,空载体组将未感染的0.1mL骨髓间充质干细胞悬液注入颈总动脉外膜,分3点注射;血管生长素1转染组同法注入等量重组慢病毒感染的骨髓间充质干细胞悬液,饲养2个月后处死大鼠,取颈动脉组织。主要观察指标:Western blot法检测转基因干细胞中血管生长素1的表达,荧光免疫组织化学及苏木精-伊红染色检测动脉内膜/中膜面积比。结果:24只大鼠均进入结果分析。血管生长素1转染组表达血管生长素1蛋白,另2组无血管生长素1蛋白表达。移植后2个月,荧光显微镜下空载体组、血管生长素1转染组动脉壁内均可见绿色荧光蛋白阳性细胞;各组均可见动脉内膜增生,斑块形成;与模型对照组比较,空载体组内膜面积/中膜面积值无明显变化(P>0.05),血管生长素1转染组内膜面积/中膜面积值明显减小(P<0.05)。结论:经血管生长素1基因修饰的骨髓间充质干细胞移植可明显减轻动脉粥样硬化程度。更多还原

【Abstract】 BACKGROUND:Bone marrow derived mesenchymal stem cells(MSCs) are easy transfected by heterologous genes,with the characteristics of chemotaxis and congregation to injured tissues.The ability to efficiently transfer genes of interest into such cells would create a number of therapeutic opportunities.Therefore,MSCs can differentiate into vascular endothelial cells(VECs) to repair injured tunica intima.OBJECTIVE:To investigate the influence of lentivirus-mediated angiopoietin-1(Ang1) gene modified MSCs on carotid atherosclerosis after transplantation.DESIGN,TIME AND SETTING:A randomized controlled experiment with animals as subjects.The experiment was carried out in the Hypertension Institute of Fujian Province and Center Laboratory of the First Affiliated Hospital of Fujian Medical University from February 2007 to May 2008.MATERIALS:Thirty-two male SD rats,4 weeks old,were selected.Eight of them were prepared for MSCs;the rest were randomly divided into control,empty vector and Ang1 transfected MSCs group,with 8 animals in each group.METHODS:Genetic engineering MSCs were constructed by lentivirally-tranduced Ang1 gene,the green fluorescent protein(GFP) was used as report gene.Regulated the cell density with(1.0-2.0) ×1010/L.Rat carotid atherosclerosis model was established by balloon injury combined with high fat diet.0.1 mL MSCs suspension was injected into the adventitia of the common carotid artery at 3 points,at immediately time after model establishment in the empty vector group;same volume of Ang1 transfected MSCs suspension was injected with same method in the Ang1 transfected MSCs group.All rats were sacrificed after 2 months,and carotid artery tissues were harvested.MAIN OUTCOME MEASURES:The expression of Ang1 in transfected MSCs was detected by Western blot,and ratio of neointimal/medial area was calculated by hematoxylin-eosin staining.RESULTS:All the 24 rats were involved in the result analysis.Only Ang1 transfected MSCs group could express Ang1 protein.Green fluorecent protein could be observed under the fluorecent microscopy in the empty vector and Ang1 transfected MSCs groups at 2 months after transplantation.The arterial neointimal hyperplasia with plaque formation was seen in each group.Compared to control group,there was not significantly difference in neointimal/medial area between the empty vector group(P > 0.05),which was significantly decreased in Ang1 transfected MSCs group(P < 0.05).CONCLUSION:The transplantation of Ang-1 gene modified MSCs can significantly relieve the severity of atherosclerosis.更多还原