Shang Feng, He Zhi-xu, Wang Hao-wen, Cui Dong-bing, Yang Yan
Department of Pediatrics, Dezhou People's Hospital of Shandong Province, Dezhou, China.
Zhonghua Yi Xue Za Zhi. 2011 Jul 12;91(26):1847-51.
To explore the therapeutic effect and the mechanism of marrow mesenchymal stem cells (MMSCs) transfected with vascular endothelial growth factor (VEGF) gene in the treatment of pulmonary hypertension in rats.
MMSCs from the bone marrow of Sprague-Dawley rats were isolated, cultured and propagated in vitro. pIRES2-EGFP-VEGF165 was transfected 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 l 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), right ventricular hypertrophy index, blood gas analysis and microstructure as well as pulmonary microvascular changes were observed after 30 days.
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 right 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 were of statistical significances (P < 0.01). A comparison of right ventricle hypertrophy index, MT and V/T was of statistical significance between MMSC and gene transplantation groups (P < 0.05). The blood gas analysis of the MMSCs transplantation and gene transplantation groups were better than the pulmonary hypertension mode group. Ultramicrostructure showed that neovascularization and small pulmonary arterial repair appeared in two transplantation groups.
MMSCs transfected by pIRES2-EGFP-VEGF165 transplantation may improve and reverse the MCT-induced progress of pulmonary hypertension in rats. And it is better than the MMSC transplantation. The potential mechanism is through arterial repair and neovascularization.
探讨转染血管内皮生长因子（VEGF）基因的骨髓间充质干细胞（MMSCs）治疗大鼠肺动脉高压的疗效及机制。
从Sprague-Dawley大鼠骨髓中分离、培养并体外扩增MMSCs。将pIRES2-EGFP-VEGF165转染至MMSC。健康雄性SD大鼠随机分为4组：正常对照组、肺动脉高压模型组、MMSCs移植组和转基因移植组。单次皮下注射野百合碱（50 mg/kg）诱导肺动脉高压模型。正常对照组单次皮下注射L-DMEM（低糖杜尔贝科改良伊格尔培养基）。4组大鼠饲养方式相同。造模后第21天，将1 ml L-DMEM中含5×10(6)个MMSCs注入MMSCs组。将5×10(5)个经pIRES2-EGFP-VEGF165转染的MMSC注入基因移植组。肺动脉高压模型组和正常对照组也注射相同体积的L-DMEM溶液。30天后观察右心室收缩压（RVSP）、右心室肥厚指数、血气分析、微观结构以及肺微血管变化等参数。
MMSCs移植后第30天，结果如下：MMSCs移植组和基因移植组的RVSP分别为（30.2±2.1）和（29.2±1.1）mmHg（1 mmHg = 0.133 kPa）。右心室肥厚指数分别为（37.±3.2）%和（27.2±3.4）%。肺动脉中膜厚度（MT）分别为（21.3±3.4）和（14.3±2.8）µm。血管面积与总动脉面积之比（V/T）分别为（39.3±4.3）%和（43.0±1.5）%。与肺动脉高压模型组相比，上述参数具有统计学意义（P < 0.01）。MMSC组和基因移植组之间右心室肥厚指数、MT和V/T的比较具有统计学意义（P < 0.05）。MMSCs移植组和基因移植组的血气分析均优于肺动脉高压模型组。超微结构显示，两个移植组均出现新生血管形成和小肺动脉修复。
经pIRES2-EGFP-VEGF165转染的MMSCs移植可改善并逆转野百合碱诱导的大鼠肺动脉高压进程，且优于MMSCs移植。潜在机制是通过动脉修复和新生血管形成。
