在啮齿动物冷冻损伤心脏模型中使用基因调控的人骨骼肌成肌细胞逆转心肌损伤

Reversal of myocardial injury using genetically modulated human skeletal myoblasts in a rodent cryoinjured heart model.

作者信息

Ye Lei, Haider Husnain Kh, Jiang Shujia, Ling Lieng H, Ge Ruowen, Law Peter K, Sim Eugene K W

机构信息

Department of Cardiothoracic and Vascular Surgery, National University of Singapore-117597, Singapore.

出版信息

Eur J Heart Fail. 2005 Oct;7(6):945-52. doi: 10.1016/j.ejheart.2005.03.012. Epub 2005 Oct 3.

DOI:10.1016/j.ejheart.2005.03.012

PMID:16202652

Abstract

BACKGROUND

We hypothesized that combination therapy using human myoblasts and VEGF165 will lead to better prognosis in a failing heart.

METHODS

Forty-eight female Wistar rats with cryoinjured hearts were randomized into non-treated normal (group-1, n=12), DMEM injected (group-2, n=10), myoblast-transplanted (group-3, n=12) and myoblast-hVEGF(165) (group-4, n=14). Ten days after cryoinjury, 200 microl DMEM containing 3x10(6) cells or without cells was injected into the injured myocardium. Animals were maintained on cyclosporine for 6 weeks post cell transplantation. Heart function was assessed by echocardiography. Animals were sacrificed and hearts were processed for histochemical and immunohistochemical studies.

RESULTS

Histological examination showed survival of the donor myoblasts expressing lac-z and hVEGF165 in rat cardiac tissue. Fluorescent immunostaining for vWillebrand Factor-VIII and smooth muscle actin expression at low power microscope (x100) showed significantly higher blood vessel density in group-4 (31.25+/-1.82; 24.63+/-0.92) as compared to group-2 (13.29+/-1.0; p<0.001; 9.71+/-0.81, p<0.001) and group-3 (16.50+/-1.43, p<0.001; 14.5+/-1.34, p<0.001). Echocardiography showed that ejection fraction and fractional shortening of group-3 (93.36+/-1.52%, p=0.005; 75+/-3.75%, p=0.024) and group-4 (94.8+/-1.62%, p=0.003; 76.13+/-2.15%, p=0.011) significantly improved as compared to group-2 (81.8+/-3.3%, 55.1+/-7.18%).

CONCLUSION

Myoblasts carrying of hVEGF165 are potential therapeutic transgene carriers for cardiac repair.

摘要

背景

我们推测，使用人成肌细胞和VEGF165进行联合治疗将使衰竭心脏的预后更好。

方法

将48只心脏冷冻损伤的雌性Wistar大鼠随机分为未治疗的正常组（第1组，n = 12）、注射DMEM组（第2组，n = 10）、成肌细胞移植组（第3组，n = 12）和成肌细胞-hVEGF(165)组（第4组，n = 14）。冷冻损伤10天后，将含有3×10(6)个细胞或不含细胞的200微升DMEM注射到损伤心肌中。细胞移植后，动物接受环孢素治疗6周。通过超声心动图评估心脏功能。处死动物，取心脏进行组织化学和免疫组织化学研究。

结果

组织学检查显示，供体成肌细胞在大鼠心脏组织中存活并表达lac-z和hVEGF165。在低倍显微镜（×100）下对血管性血友病因子VIII和平滑肌肌动蛋白表达进行荧光免疫染色显示，与第2组（13.29±1.0；p<0.001；9.71±0.81，p<0.001）和第3组（16.50±1.43，p<0.001；14.5±1.34，p<0.001）相比，第4组的血管密度显著更高（31.25±1.82；24.63±0.92）。超声心动图显示，与第2组（81.8±3.3%，55.1±7.18%）相比，第3组（93.36±1.52%，p = 0.005；75±3.75%，p = 0.024）和第4组（94.8±1.62%，p = 0.003；76.13±2.15%，p = 0.011）的射血分数和缩短分数显著改善。