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中华细胞与干细胞杂志(电子版) ›› 2017, Vol. 07 ›› Issue (05) : 271 -276. doi: 10.3877/cma.j.issn.2095-1221.2017.05.004

所属专题: 文献

论著

碱性成纤维生长因子转染骨髓间充质干细胞移植对心肌病心力衰竭大鼠心功能的影响
赖增华1,(), 窦学凯1, 贾春文1, 姜朝晖1   
  1. 1. 360001 厦门,中国人民解放军第一七四医院(厦门大学附属成功医院)心内科
  • 收稿日期:2017-05-19 出版日期:2017-10-01
  • 通信作者: 赖增华

Influence of transplantation with basic fibroblast growth factor gene-transfected bone mesenchymal stem cells on heart function in rats with heart failure

Zenghua Lai1,(), Xuekai Dou1, Chunwen Jia1, Chaohui Jiang1   

  1. 1. Department of Cardiology, the 174th Hospital of the PLA, Affiliated Chenggong Hospital of Xiamen University, Xiamen 360001, China
  • Received:2017-05-19 Published:2017-10-01
  • Corresponding author: Zenghua Lai
  • About author:
    Corresponding author: Lai Zenghua, Email:
引用本文:

赖增华, 窦学凯, 贾春文, 姜朝晖. 碱性成纤维生长因子转染骨髓间充质干细胞移植对心肌病心力衰竭大鼠心功能的影响[J/OL]. 中华细胞与干细胞杂志(电子版), 2017, 07(05): 271-276.

Zenghua Lai, Xuekai Dou, Chunwen Jia, Chaohui Jiang. Influence of transplantation with basic fibroblast growth factor gene-transfected bone mesenchymal stem cells on heart function in rats with heart failure[J/OL]. Chinese Journal of Cell and Stem Cell(Electronic Edition), 2017, 07(05): 271-276.

目的

探讨碱性成纤维生长因子(bFGF)转染的骨髓间充质干细胞(BMSCs)移植对心肌病心力衰竭大鼠心功能的影响。

方法

通过腹腔注射阿霉素建立心肌病心力衰竭SD大鼠模型,存活33只大鼠随机分成3组:模型组、BMSCs组和BMSCs+bFGF组,每组11只。另设正常对照组(n = 8)。取体外培养的第三代SD大鼠BMSCs,质粒-bFGF转染BMSCs,BrdU标记后,分别将BMSCs(3×106个)、bFGF转染的BMSCs(3×106个)或等体积培养基通过尾静脉注射的方法进行移植。用生物信号采集系统检测大鼠心功能,大鼠心脏切片行免疫组化了解移植细胞在受体心脏的存活情况,实时荧光定量PCR和Western Blotting检测心室组织bFGF的表达水平。多组间比较采用单因素方差分析或精确概率法。

结果

BMSCs移植4周后,正常对照组大鼠的心功能指标LVSP(121.13±12.28)mmHg,LVDEP(3.86±1.25)mmHg,LV+dP/dt(3671.25±172.50)mmHg/s和LV-dP/dt(3221.63±259.57)mmHg/s;模型组心功能指标LVSP(81.29±12.39)mmHg,LVDEP(16.43±4.12)mmHg,LV+dP/dt(2344.29?±98.63)mmHg/s和LV-dP/dt(2244.29±103.10)mmHg/s;BMSCs组心功能指标LVSP(97.00?±6.39)mmHg,LVDEP(12.00±2.73)mmHg,LV+dP/dt(2876.25±118.31)mmHg/s和LV-dP/dt(2726.25?±?303.23)mmHg/s;BMSCs+bFGF组心功能指标LVSP(109.38±12.78)mmHg,LVDEP(9.25?±1.67)mmHg,LV+dP/dt(3037.50±161.22)mmHg/s和LV-dP/dt(3000.13±149.77)mmHg/s,移植组各指标均有改善,比较差异有统计学意义(F = 17.29、36.61、111.11、26.54,P均< 0.01)。免疫组织化学检查示细胞移植组大鼠的心室组织切片上可见BrdU阳性细胞存在。模型组大鼠心室心肌组织bFGF mRNA表达水平高于正常对照组(1.28±0.06vs1.00±0.00,t = 10.46,P < 0.01),BMSCs组高于模型组(1.37±0.05vs1.28±0.06,t=3.39,P < 0.01),而BMSCs+bFGF组高于BMSCs组(1.44±0.03vs1.37±0.05,t =2.86,P = 0.01)。Western Blotting示,正常对照组大鼠心室组织bFGF蛋白表达最低,模型组高于正常对照组(1.31±0.09vs1.00?±0.00,t = 5.48,P = 0.01),BMSCs组高于模型组(1.46±0.05vs1.31±0.09,t = 2.68,P = 0.03),而BMSCs+bFGF组高于BMSCs组(1.84±0.09vs1.46±0.05,t = 6.79,P < 0.01)。

结论

转染bFGF基因的BMSCs移植可提高阿霉素所致心肌病心力衰竭大鼠心脏的bFGF水平,改善大鼠心功能,其效果优于单纯BMSCs移植。

Objective

To explore the influence of transplantation with basic fibroblast growth factor (bFGF) gene-transfected bone mesenchymal stem cells (BMSCs) on heart function in rats with heart failure.

Methods

A heart failure model of SD rats was induced by intraperitoneal injection with adriamycin. The 33 survived rats were randomly divided into 3 groups: a model group, a BMSCs group and a BMSCs+bFGF group (n= 11 for all). Another 8 rats served as control. BMSCs at the third passage were transfected with bFGF and labeled with BrdU. BMSCs (3×106), bFGF-transfected BMSCs (3×106), or culture medium were respectively transplanted into rats via the tail vein. Cardiac function was evaluated with the use of a physiological recorder. The hearts were analyzed by immunohistochemical staining to identify the transplanted BrdU-labeled cells. The expressions of bFGF protein and mRNA in the heart tissues were respectively determined by Western Blotting and real time PCR. One-Way ANOVA or Fisher's exact test was used in the analysis as appropriately.

Results

Four weeks after transplantation, hemodynamic measurements showed LVSP (121.13±12.28) mmHg, LVDEP (3.86±1.25) mmHg, LV +dP/dt (3671.25±172.50) mmHg/s, LV-dP/dt (3221.63±259.57) mmHg/s in the normal control group; LVSP (81.29±12.39) mmHg, LVDEP (16.43±4.12) mmHg, LV+dP/dt (2344.29±98.63) mmHg/s, LV-dP/dt (2244.29±103.10) mmHg/s in the model group; LVSP (97.00±6.39) mmHg, LVDEP (12.00±2.73) mmHg, LV+dP/dt (2876.25±118.31) mmHg/s, LV-dP/dt (2726.25±303.23) mmHg/s in the BMSCs group, and LVSP (109.38±12.78) mmHg, LVDEP (9.25±1.67) mmHg, LV+dP/dt (3037.50±161.22) mmHg/s, LV-dP/dt (3000.13±149.77) mmHg/s in the BMSCs+bFGF group. The parameters were all significantly improved (F= 17.29, 36.61, 111.11 and 26.54 respectively, Pall < 0.01) after the transplantation. BrdU-labeled transplanted BMSCs could be found in the hearts of the recipients. The expression of bFGF mRNA in the heart tissues were significantly increased in the model group (1.28±0.06) than in the control group (t= 10.46, P< 0.01); in the BMSCs group (1.37±0.05) than in the model group (t?= 3.39,P< 0.01); and in the BMSCs+bFGF group (1.44±0.03) than in the BMSCs group (t=2.86,P= 0.01). As shown by Western Blotting test, the bFGF protein expression level of the heart tissues was the lowest in the normal control, but significantly higher in the model group than in the control group (1.31±0.09vs1.00±0.00,t= 5.48,P= 0.01), in the BMSCs group than in the model group (1.46?±0.05vs1.31±0.09,t= 2.68,P= 0.03), and in the BMSCs+bFGF group than in the BMSCs group (1.84±0.09vs1.46±0.05,t= 6.79,P< 0.01).

Conclusion

Transplantation with BMSCs or bFGF-transfected BMSCs can both improve cardiac function in rats with adriamycin-induced heart failure, while the effect of bFGF-transfected BMSCs may be better than BMSCs only.

表1 各组大鼠经BMSCs移植术后第4周左室功能比较(±s)
图1 光学显微镜下观察大鼠心脏心脏切片的BrdU免疫组化(SABC,×400)
图2 Western Blotting法检测BMSCs移植对大鼠心室组织bFGF蛋白表达的影响
图3 Western Blotting法检测BMSCs移植对大鼠心室组织bFGF蛋白表达的影响
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