血管内皮生长因子基因转染的骨髓间充质干细胞移植对大鼠肺动脉高压的治疗作用  

Transplantation of marrow mesenchymal stem cells transfected with vascular endothelial growth factor gene for the treatment of pulmonary hypertension in rats

作　　者：尚峰[1] 何志旭[2] 汪浩文[3] 崔冬冰[2] 杨燕[2] 

机构地区：[1]山东省德州市人民医院儿科,253000 [2]贵阳医学院附属医院儿科 贵阳医学院干细胞研究中心 [3]安徽省池州市人民医院儿科

出　　处：《中华医学杂志》2011年第26期1847-1851,共5页National Medical Journal of China

基　　金：国家自然科学基金（30560159）;教育部新世纪优秀人才支持计划项目（NCET-05-0820）;贵州省科技计划课题（黔科合SY字2008-3026号）

摘　　要：目的探讨血管内皮生长因子（VEGF）基因转染的骨髓间充质干细胞（MMSC）移植治疗肺动脉高压（PH）的疗效和机制。方法体外分离培养SD大鼠MMSC，将含有增强型绿色荧光蛋白的VEGF质粒转进MMSC。将SD大鼠随机分为4组：正常对照组、PH模型组；MMSC移植组，转基因移植组。一次性项背部皮下注射野百合碱（50mg,／kg），复制PH模型，观察21d后，MMSC移植组及转基因移植组分别给予浓度为5×10^5个细胞／ml和5×10^5个细胞／ml的MMSC悬液1ml静脉注射，模型组及正常组均给予等量L-DMEM液注射。移植后30d，观察4组大鼠一般情况、生存率、右心室收缩压（RVSP）、右心室肥大指数（RVH）、血气及肺小动脉的微观结构变化。结果移植后30d，MMSC移植组及转基因移植组大鼠的RVSP分别为（30．2±2．1）和（29．2±1．1）mmHg（1mmHg=0．133kPa），RVH分别为（37．9±3．2）％和（27．2±3．4）％，肺小动脉中膜厚度（MT）分别为（21．3±3．4）和（14．3±2．8）μm，肺小动脉管腔横截面积与血管总横截面积的比值（V／T）分别为（39．3±4．3）％和（43．0±1．5）％。上述指标与PH模型组相比差异均有统计学意义（P〈0．01）；其中MMSC移植组与转基因移植组间RVH、MT及V／T的差异有统计学意义（P〈0．05）。另外，MMSC移植组与转基因移植组动脉血气各指标较PH模型组均明显改善（P〈0．05）。同时观察到肺内损伤血管修复和血管新生现象。结论转染有VEGF165基因的MMSC移植可有效减轻并逆转野百合碱诱导的PH进程，此作用较单纯移植MMSC的治疗效果更加明显。其作用机制可能与血管修复及血管新生有关。Objective To explore the therapeutic effect and the mechanism of marrow mesenchymal stem cells （MMSCs） transfeeted with vascular endothelial growth factor （VEGF） gene in the treatment of pulmonary hypertension in rats. Methods MMSCs from the bone marrow of Sprague-Dawley rats were isolated, cultured and propagated in vitro, pIRES2-EGFP-VEGF165 was transfeeted into MMSC. The healthy male SD rats were divided randomly into 4 groups： normal control group, pulmonary hypertension model group, MMSCs transplantation group and transfer gene transplantation group. A single subcutaneous monocrotaline （50 mg/kg） was injected to induce the model of pulmonary hypertension. The normal control group received a single subcutaneous dose of L-DMEM （low glucose Dulbecco＇s modified Eagle＇s medium）. All four groups of rats were fed similarly. At Day 21 post-modeling, 5 ×10^6 MMSCs in 1 ml L-DMEM were injected into the MMSC group. 5 ×10^5 MMSC transfected by pIRES2-EGFP-VEGF165 were injected into the gene transplantation group. A same volume L-DMEM solution was also injected into the pulmonary hypertension model group and normal control group. The parameters of right ventricular systolic pressure （RVSP）, fight ventrieular hypertrophy index, blood gas analysis and microstructure as well as pulmonary microvascular changes were observed after 30 days. Results At Day 30 post-transplantation of MMSCs, the outcomes were as follows： RVSP was （30. 2 ±2. 1 ） and （29. 2 ±1.1 ）mm Hg （ 1 mm Hg =0. 133 kPa） in the MMSCs transplantation and gene transplantation groups respectively. The fight ventricular hypertrophy indices were （ 37.9 ±3.2） % and （ 27.2 ±3.4 ） % respectively. The media thickness of pulmonary artery （MT） was （ 21.3 ±3.4 ） and （ 14. 3 ±2. 8 ） μm respectively. The ratios of vascular area to total arterial area （ V/T ） were （ 39. 3 ±4. 3 ） % and （ 43.0 ±1.5 ） % respectively. As compare with the pulmonary hypertension model group, the above parameters wer

关 键 词：高血压 肺性 间质干细胞移植 血管内皮生长因子类 基因 

分 类 号：R543.2[医药卫生—心血管疾病]