人肝细胞生长因子基因修饰的骨髓间充质干细胞对大鼠心肌纤维化的影响

The effect of human hepatocyte growth factor gene-modified bone marrow mesenchymal stem cells on myocardial fibrosis in rats

目的观察携带人肝细胞生长因子基因修饰的骨髓间充质干细胞(MSC-hHGF)对心力衰竭模型大鼠心肌纤维化影响。方法 50只12周龄清洁级雄性SD大鼠[体质量(200±10)g]随机抽取14只作为对照组,其余36只大鼠腹腔注射盐酸阿霉素制作心力衰竭模型,存活大鼠(n=24)再随机平分为3组:模型组、骨髓间充质干细胞(MSC)组和MSC-hHGF组。采用密度梯度离心-贴壁培养法获取SD大鼠的MSC,Ad-hHGF转染HGF到MSC,ELISA检测hHGF在该细胞的表达情况。腹腔注射阿霉素建立扩张型心肌病心力衰竭大鼠模型。细胞移植后4周,用生物信号采集系统检测心功能,免疫组化检测左室心肌组织HGF表达水平,天狼猩红染色评价心肌纤维含量,偏振光显微镜分析瘢痕区Ⅰ型胶原表达情况。结果与模型组相比,MSC组的左室收缩峰压(LVSP)[(41.1±3.7)比模型组(33.2±1.4)mmHg,P<0.05]升高,MSC组的左室舒张末期压(LVEDP)[(15.7±1.3)比模型组(19.5±2.1)mmHg,P<0.05]降低;与MSC组相比,MSC-hHGF组LVSP[(49.6±6.1)比MSC组(41.1±3.7)mmHg,P<0.05]升高;MSC-hHGF组LVEDP[(8.7±1.1)比MSC组(15.7±1.3)mmHg,P<0.05]降低。MSC-hHGF组心肌组织Ⅰ型胶原含量与模型组比降低,但是与对照组比没明显差异。结论 MSC-hHGF可明显改善心力衰竭大鼠心功能和心肌纤维化。

Objective To observe the effect of human hepatocyte growth factor gene-modified bone marrow mesenchymal stem cells on myocardial fibrosis in the rats with heart failure.Methods A total of 50 male SD rats [12-week-old,weight（200±10）g] were included,of which 14 rats were randomly selected and served as control.A model of heart failure was induced by intraperitoneal injection of doxorubicin hydrochloride in the remaining rats（n=36）.The survivals of rats（n=24）were randomly divided into three groups,the model group,the bone marrow mesenchymal stem cell transplantation group（the MSC group）and the hepatocyte growth factor gene transfected MSC transplantation group（the MSC-hHGF group）.MSC were isolated and purified by the means of density gradient centrifugation and adherence,HGF was transfected to MSC by Ad-hHGF,expression of hHGF of MSC was detected by ELISA,the model of heart failure was induced by intraperitoneal injection of doxorubicin hydrochloride.Four weeks after cell transplantation,cardiac function was evaluated using physiological recorder,expression of hHGF of implanted MSC in left ventricle was evaluated by immunohistochemistry staining,the myocardial fiber content was assessed by sirius red staining,and collagen type Ⅰ expression of the scar area was analysed by polarized light microscopy.Results Compared with model group,left ventricular systolic pressure（LVSP）of MSCs group were higher [LVEDP,MSCs（41.1±3.7）vs model group（33.2±1.4）mm Hg,P〈0.05],left ventricular end-diastolic pressure（LVEDP）of MSCs group were lower [LVEDP,MSCs（15.7±1.3）vs model group（19.5±2.1）mm Hg,P〈0.05];Compared with MSCs group,LVSP of MSCs-hHGF group were higher[LVEDP,MSCs-hHGF（49.6±6.1）vs MSCs group（41.1±3.7）mm Hg,P〈0.05],LVEDP of MSCs-hHGF group were lower [LVEDP,MSC-hHGF（8.7±1.1）vs MSC group（15.7±1.3）mm Hg,P〈0.05].Collagen type 1 of MSC-hHGF group were lower than model group,but have no difference with control group.Conclusion Compared with MSC,hHGF-modified bone marrow mesenchymal stem cell transplantation can highly improve left ventricular function and myocardial fibrosis in the rats with heart failure.