携带肝细胞生长因子基因的骨髓间充质干细胞移植对肺动脉高压大鼠治疗作用初探

目的 研究携带肝细胞生长因子(hepatocyte growth factor,HGF) 基因的骨髓间充质干细胞(mesenchymal stem cells,MSCs) 移植对肺动脉高压大鼠血流动力学指标的变化.方法 取3 周龄雄性SD 大鼠骨髓体外培养并纯化MSCs,利用腺病毒(adenovirus,Ad) 介导的基因转染方法,将HGF 基因导入MSCs.将SD 大鼠30 只随机分为3 组(n=10):肺动脉高压组(PAH,P 组):采用腹腔注射野百合碱(60mg/kg) 复制PAH 模型后,经颈内静脉移植1ml 低糖伊戈尔培养基(L-DMEM) ;携带空腺病毒载体骨髓间充质干细胞移植组(MSCs,M 组):复制PAH 模型后,移植5×106/L 空腺病毒载体转染的MSCs 细胞悬液1ml ;腺病毒介导HGF 基因转染的骨髓间充质干细胞移植组(HGF,H 组):复制PAH 模型后,移植5×106/L 腺病毒介导HGF 基因转染的MSCs 细胞悬液1ml.观察各组大鼠血流动力学变化、右心室肥大指数(RV/LV 比值) 以及血浆中ET-1的水平.结果 肺动脉高压大鼠治疗3 周后,实验各组动脉血压(BP) 无明显差异;H 组肺动脉收缩期压力(pulmonary arterysystdic pressure,PASP) 和平均肺动脉压(mean pulmonary arterial pressure,mPAP)(37.227 2±5.381 60,25.693 5±1.446 33)均较P 组(45.069 0±7.177 97,30.800 2±1.256 15) 和M 组(44.142 0±6.673 75,30.359 1±1.172 52) 显著下降(P<0.05) ;H组RV/LV 比值(42.200 0±8.060 59) 较P 组(46.858 0±8.241 61) 和M 组(46.750 0±10.420 14) 有所下降,但无统计学差异;H 组血浆中ET-1 含量(23.862 70±7.426 76) 较P 组(28.372 0±4.402 75) 和M 组(31.849 0±7.547 13) 均有所下降,但仅较M 组有统计学意义(P<0.05).结论 携带HGF 基因的骨髓间充质干细胞移植治疗法可在不影响全身血流动力学基础上,显著降低肺动脉高压大鼠的肺动脉收缩压和肺动脉平均压,改善大鼠肺血管的血流动力学,并降低血浆中内皮素-1 含量,从而起到保护血管内皮细胞、拮抗血管收缩作用.

Role of bone marrow mesenchymal stem cells transplantation in treatment of pulmonary arterial hypertension in rats carrying hepatocyte growth factor

Objective To study the role of bone marrow mesenchymal stem cells(MSCs) transplantation in treatment of monocrotaline(MCT)-induced pulmonary arterial hypertension(PAH) in rats carrying hepatocyte growth factor(HGF) and its mechanism. Methods MSC isolated from bone marrow of 3-week old male SD rats were incubated in vitro with adhesion culture method,purified and transfected with adenovirus(Ad)-mediated HGF.The rats were randomly divided into PAH group,MSC transplantation group,and Ad-mediated HGF transplantation group,10 in each group.After a PAH model was established by intraperitoneal injection of monocrotaline(MCT) at the dose of 60mg/kg,1ml L-DMEM was injected into the jugular vein of rats in PAH group,1ml suspension containing 5×106 MSC transfected with empty adenovirus vector was injected into rats of HGF transplantation group.Hemodynamics,RV/LV and plasma ET-1 level in rats of different groups were observed.Results No significant difference was found in BP among different groups 3 weeks after treatment of PAH(pulmonary arterial hypertension).The PASP(pulmonary artery systdic pressure) and mPAP(mean pulmonary arterial pressure) were significantly lower in Ad-mediated HGF transplantation group than in PAH group and MSCs transplantation group(37.227 2±5.381 60 vs 45.069 0±7.177 97 and 44.142 0±6.673 75,25.693 5± 1.446 33 vs 30.800 2±1.256 15 and 30.359 1±1.172 52,P<0.05).No significant difference was found in RV/LV among Ad-mediated HGF transplantation group,PAH group and MSCs transplantation group(42.200 0±8.060 59,46.858 0±8.241 61,46.750 0±10.420 14, P>0.05)although it was decreased.The plasma ET-1 level was significantly lower in Ad-mediated HGF transplantation group than in PAH group and MSC transplantation group(23.862 70±7.426 76 vs 28.372 0±4.402 75 and 31.849 0±7.547 13,P<0.05).Conclusion MSC carrying HGF transplantation therapy can significantly reduce the PASP and mPAP,improve hemodynamics,delay and even reverse right ventricular hypertrophy,and decrease plasma ET-1 level in rats with PAH,thus protecting vascular endothelium cells against damage and blood vessels against constriction.