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目的:观察以骨髓干细胞(BMSCs)介导的人肝细胞生长因子(hHGF)基因转染在兔心肌梗死组织中能否稳定表达。方法:家兔16只,复制急性心肌梗死(AMI)模型。随机分为2组:移植组:BMSCs加hHGF移植,对照组。另外随机选取8只兔作为假手术组。AMI术后3 d,抽取股骨骨髓1.5 ml,分离BMSCs培养。于对数生长期加入5-溴脱氧尿苷孵育24 h,以标记移植细胞。构建pcDNA3.1-hHGF重组表达载体,移植前用脂质体包裹表达载体转染BMSCs,孵育24 h。AMI术后14 d,行自体细胞移植,对照组接受等量的无血清培养基注射,假手术组只开胸,不注射。细胞移植后28 d,检测左心室血流动力学指标;取心肌组织,RT-PCR检测hHG-FmRNA的表达;酶联免疫吸附法(ELISA)测定心肌组织中HGF的含量。结果:与假手术组比较,心肌梗死后28d,对照组左心室功能降低,但BMSCs加hHGF组左心心室功能有改善;RT-PCR结果显示移植组有hHGFmR-NA表达,ELISA测定HGF含量移植组高于对照组。结论:AMI组织中HGF含量降低,转染HGF的BMSCs能在AMI组织中生存并能表达分泌HGF,其左心室功能有改善。
OBJECTIVE: To observe whether stable transfection of human hepatocyte growth factor (hHGF) gene is mediated by bone marrow stem cells (BMSCs) in rabbit myocardial infarction. Methods: Twenty-six rabbits were used to replicate acute myocardial infarction (AMI) model. Randomly divided into two groups: transplantation group: BMSCs plus hHGF transplantation, control group. In addition, 8 rabbits were randomly selected as sham operation group. Three days after AMI, 1.5 ml femur bone marrow was collected and BMSCs were isolated and cultured. In logarithmic growth phase, 5-bromodeoxyuridine was incubated for 24 h to label the transplanted cells. Recombinant pcDNA3.1-hHGF expression vector was constructed and transfected into BMSCs by liposome-encapsulated expression vector for 24 h before transplantation. At 14 days after AMI, autologous cells were transplanted. The control group received the same amount of serum-free medium injection. Sham-operated group received thoracotomy without injection. Thirty-eight days after transplantation, left ventricular hemodynamic parameters were measured. Myocardial tissues were harvested for detection of hHG-F mRNA expression by RT-PCR. HGF level in myocardium was measured by enzyme-linked immunosorbent assay (ELISA). Results: Compared with the sham operation group, the left ventricular function decreased in the control group 28 d after myocardial infarction. However, the left ventricular function was improved in the BMSCs plus hHGF group. The expression of hHGFmR-NA in the transplantation group was detected by RT-PCR. Group higher than the control group. CONCLUSIONS: HGF levels in AMI tissues are decreased. BMSCs transfected with HGF can survive and express HGF in AMI tissues with improved left ventricular function.