hHGF基因修饰对MSCs移植改善重度肺动脉高压大鼠肺微血管稀薄作用的影响  

Effect of human hepatocyte growth factor genetic modification on the ameliorating effects of MSCs implantation on pulmonary microvascular rarefaction in a rat model of pulmonary hypertension

作　　者：林群[1] 雷立华[2] 林财珠[1] 曾邦雄[3] 梁富球[1] 林献忠[1] 郑辉哲[4] 蔡宏达[1] 高友光[1] 杨庆[1] 

机构地区：[1]福建医科大学附属第一医院麻醉科,福州市350005 [2]福建医科大学省立临床医学院麻醉科 [3]华中科技大学同济医学院附属协和医院麻醉科 [4]福建省肿瘤医院麻醉科

出　　处：《中华麻醉学杂志》2012年第10期1252-1256,共5页Chinese Journal of Anesthesiology

基　　金：国家自然科学基金（30972972）;福建省自然科学基金（2011J01129）;福建省医学创新课题（2009-CXB-4）

摘　　要：目的评价人肝细胞生长因子（hHGF）基因修饰对骨髓间质干细胞（MSCs）移植改善重度肺动脉高压大鼠肺微血管稀薄作用的影响。方法将原代培养的F344大鼠MSCs转导携带HGF基因的慢病毒载体和空载体，以获得HGF—MSCs和EGFP—MSCs。7周龄近交系雄性F344大鼠，体重180～250g，采用野百合碱复合单肺切除法建立重度肺动脉高压大鼠模型。取重度肺动脉高压大鼠66只，采用随机数字表法，将其随机分为3组（n=22）：对照组（C组）、空载体慢病毒转导的MSCs（EGFP-MSCs）组（E组）和hHGF转导的MSCs（hHGF—MSCs）组（H组）。c组经颈外静脉注射DMEM1ml，E组注射含5×10^5个EGFP．MSCs的DMEM1ml和H组注射含5×10^5个hHGF．MSCs的DMEM1ml。注射野百合碱后45d内进行生存分析；颈外静脉注射后2周，记录平均肺动脉压（MPAP），计算右心室肥厚指数，观察肺组织MSCs分布情况，测定肺组织鼠肝细胞生长因子（rHGF）和hHGF蛋白的含量，测定Ⅷ因子的表达情况以计算肺微血管密度，并行肺动脉钡剂灌注造影。结果C组和E组肺组织均未检测出hHGF。与C组比较，E组和H组MPAP和右心室肥厚指数降低，肺组织rHGF蛋白含量、肺微血管密度和生存率升高（P〈0．01）。与E组比较，H组MPAP和右心室肥厚指数降低，肺组织rHGF蛋白含量、肺微血管密度和生存率升高（P〈0．01）。E组和H组肺内均可见大量绿色荧光的MSCs分布。肺动脉钡剂灌注造影显示，H组末梢血管灌注背景明显多于E组和C组。结论hHGF基因修饰可促进MSCs移植改善重度肺动脉高压大鼠肺微血管稀薄的作用。Objective To investigate the effect of human hepatocyte growth factor （hHGF） genetic modification on the ameliorating effects of mesenchymal stem cells （MSCs） implantation on pulmonary microvascular rarefaction in a rat model of pulmonary hypertension （ PH）. Methods MSCs were obtained from F344 rats and transduced with lentiviral vector modified with human HGF （hHGF-MSCs） or empty vector （EGFP-MSCs） . Sixty-six 7 week old male F344 rats weighing 180-250 g were used in this study. PH was induced by left pneumonectomy and subcutaneous monocrotaline （MCT） 60 mg/kg injected at 2 weeks after operation. The animals with PH were randomly divided into 3 groups： control group （group C）, EGFP-MSCs group （group E） and HGF-MSCs group（ group H）. Groups H and E received hHGF-MSCs or EGFP-MSCs 5 x 105 in DMEM 1 ml iv at 3 weeks after subcutaneous MCT injection, while group C received plain DMEM 1 ml. Mean pulmonary arterial pressure （mPAP） was measured and right ventricular hypertrophy and angiogenesis in the lung were assessed and the content of rat HGF （rHGF） and hHGF protein in lung tissue and pulmonary capillary dens!ty （by immuno-histochemistry） was measured at 2 weeks after MSCs implantation. The survival rates within 45 days after MCT administration were compared among the 3 groups.Results No hHGF was detected in groups C and E. Both hHGF-MSCs and EGFPMSCs significantly reduced MPAP and right ventricular hypertrophy and increased pulmonary capillary density and survival rates in groups H and E as compared with group C and the efficacy of hHGF-MSCs was significantly greater than that of EGFP-MSCs. Barium angiography revealed that distal pulmonary vasculature was significantly increased in group H as compared with groups E and C. The survival of the rats receiving hHGF-MSCs was significantly longer in group H than that in groups E and C. Conclusion hHGF genetic modification can improve the ameliorating effects of MSCs implantation on PH-related microvascular rarefaction.

关 键 词：肝细胞生长因子 间质干细胞移植 生物 基因修饰 高血压 肺性 

分 类 号：R543.2[医药卫生—心血管疾病]