骨髓干细胞介导的人肝细胞生长因子基因转染在兔心肌梗死组织中的表达

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hHGFmRNA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hHGFmRNA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.

Expression of human hepatocyte growth factor gene in infarct myocardium in rabbit by bone marrow stem cells transplantation

Objective:To observe the express of hepatocyte growth factor(HGF) after transplating the bone marrow stem cells(BMSCs)transfected with human HGF gene in infarct myocardium.Method:Sixteen AMI rabbits were randomly divided into two groups:BMSCs hHGF treated group and control group.There were eight rabbits selected randomly as the sham operation group.The 1.5 ml marrow was aspirated at the third day after operation and BMSCs were isolated and cultured in vitro.pcDNA3.1-hHGF recombinant plasmid was constructed.Liposome transfection method was used to transfect the BMSCs according to preceded designed protocol.Subsequently,the BMSCs were transplanted to the ischemic zone of the hearts at the 14th day while in control group only serum-free medium were injected in the same way.Left ventricle hemodynamic parameters were detected and hHGF mRNA express was determined by RT-PCR.HGF content in the local infarct myocardium was also tested by ELISA.Result:RT-PCR analysis demonstrated that the BMSCs contained hHGF transplantation was capable to increase the level of hHGF gene expression on the 28 th day after cell transplantation.Compared with sham group,the LVSP and dp/dtmax were significantly lowered in the transplant group and control group,whereas these parameters in the transplant group were improved than that in control group.The content of HGF in infarct myocardium was significantly decreased.but BMSCs contained hHGF transplantation could increase the level of HGF.Conclusion:The content of HGF in infarct myocardium was decreased.BMSCs transfected with human HGF gene could survive in the infarct myocardium and secret HGF as well as improve the heart function.