Gao Feng, He Tao, Wang HongBing, Yu ShiQiang, Yi DingHua, Liu WeiYong, Cai ZhenJie
Xijing Hospital, Fourth Military Medical University, ShaanXi, China.
Can J Cardiol. 2007 Sep;23(11):891-8. doi: 10.1016/s0828-282x(07)70845-0.
Treatment of ischemic heart disease (IHD) remains a worldwide problem. Gene therapy, and recently, cell transplantation, have made desirable progress. A combination of appropriate stem cells and angiogenic genes appears promising in treating IHD.
To study the results of angiogenesis and myogenesis induced by transplantation of the adenovirus carrying human vascular endothelial growth factor 165 (Ad-hVEGF(165))-transfected mesenchymal stem cells (MSCs) in IHD compared with direct MSC transplantation or Ad-hVEGF165 delivery.
Cultured MSCs were transfected by Ad-hVEGF(165), and secreted VEGF was measured by ELISA in vitro. Ad-hVEGF(165)-transfected MSCs (MSC/VEGF group), MSCs (MSC group), Ad-hVEGF(165) (VEGF group) or a serum-free medium (control group) was injected into syngeneic Wistar rats immediately after left coronary artery occlusion. All cells were marked with CM-DiI (Molecular Probes, USA) before transplantation. One week after treatment, messenger RNA expression of hVEGF(165) in the MSC/VEGF group was found to be significantly higher than in other groups by reverse transcriptase-polymerase chain reaction analysis. One month after cell transplantation, left ventricular (LV) ejection fraction, capillary density of the infarcted region, infarct size and hemodynamic parameters (including LV end-diastolic pressure, LV+dP/dt and LV-dP/dt) were measured and immunohistochemical analysis was performed.
A high level of VEGF was expressed by Ad-hVEGF(165)-transfected MSCs. LV ejection fraction, mean capillary density of the infarcted region and hemodynamic parameters were significantly improved in the MSC/VEGF group compared with the MSC group, the VEGF group and the control group (P<0.001 for all). Partly transplanted MSCs showed the cardiomyocyte phenotype, expressed desmin and cardiac troponin T, and resulted in angiogenesis in the ischemic myocardium. However, a few transplanted MSCs incorporated into the vascular structure and most of the new vascular components were host-derived.
The combined strategy of MSC transplantation and VEGF gene therapy can produce effective myogenesis and host-derived angiogenesis, resulting in the prevention of progressive heart dysfunction after myocardial infarction.
缺血性心脏病(IHD)的治疗仍是一个全球性问题。基因治疗以及最近的细胞移植都取得了理想的进展。适当的干细胞与血管生成基因相结合在治疗IHD方面似乎很有前景。
与直接进行间充质干细胞(MSC)移植或给予Ad-hVEGF165相比,研究携带人血管内皮生长因子165(Ad-hVEGF(165))转染的间充质干细胞移植诱导缺血性心脏病血管生成和心肌生成的效果。
用Ad-hVEGF(165)转染培养的间充质干细胞,体外通过酶联免疫吸附测定法检测分泌的VEGF。左冠状动脉闭塞后立即将Ad-hVEGF(165)转染的间充质干细胞(MSC/VEGF组)、间充质干细胞(MSC组)、Ad-hVEGF(165)(VEGF组)或无血清培养基(对照组)注射到同基因的Wistar大鼠体内。移植前所有细胞均用CM-DiI(美国Molecular Probes公司)标记。治疗1周后,通过逆转录聚合酶链反应分析发现MSC/VEGF组中hVEGF(165)的信使核糖核酸表达明显高于其他组。细胞移植1个月后,测量左心室(LV)射血分数、梗死区域的毛细血管密度、梗死面积和血流动力学参数(包括左心室舒张末期压力、左心室+dP/dt和左心室-dP/dt),并进行免疫组织化学分析。
Ad-hVEGF(165)转染的间充质干细胞表达高水平的VEGF。与MSC组、VEGF组和对照组相比,MSC/VEGF组的左心室射血分数、梗死区域的平均毛细血管密度和血流动力学参数显著改善(所有P<0.001)。部分移植的间充质干细胞表现出心肌细胞表型,表达结蛋白和心肌肌钙蛋白T,并在缺血心肌中导致血管生成。然而,少数移植的间充质干细胞整合到血管结构中,且大多数新的血管成分是宿主来源的。
MSC移植与VEGF基因治疗相结合的策略可产生有效的心肌生成和宿主来源的血管生成,从而预防心肌梗死后进行性心脏功能障碍。