Cardiopulmonary Molecular Biology Laboratory, Erasmus MC, Rotterdam, The Netherlands.
Cell Biochem Biophys. 2013 Nov;67(2):305-16. doi: 10.1007/s12013-013-9710-9.
Tetralogy of Fallot (ToF) is a cyanotic congenital heart disease with prominent right ventricular hypertrophy (RVH) associated with impaired myocardial oxygen and nutrient supply. Consequently, the right ventricle may manifest in altered molecular phenotype with a number of adaptive and inherited gene profiles which are largely unknown. The aim of the present study was to investigate the myocardial differential gene expression profile and to assess myocardial vascularisation in patients with ToF. DNA microarray analysis on right ventricular biopsies from ToF-patients operated for primary corrective surgery (referred as ToF-1; n = 12, mean age 0.5 year) and age matched controls (n = 6) was validated by Northern hybridisation and RT-PCR. Employing immunohistochemistry and video image analysis expression of vascular endothelial growth factor (VEGF), vascular density (by α-SMA and CD31 staining) and myocyte cross sectional area (Gomori's reticuline staining) were assessed in ToF-1 and adult patients (referred as ToF-2, n = 12, mean age 30 years) who underwent surgery for pulmonary regurgitation and compared the data with respective age matched controls (n = 6/12). DNA microarray analysis revealed altered expression pattern for 236 genes including enhanced (1.5-2.2-fold) expression of angiogenic factors and their receptors including; VEGF, flt-1, flk-1 angiopoietin-2, FGF-2, FGF-R1, PDGF-A, whereas, flt-4, Tie, TGF-β, TGF-β3R showed decreased (1.6-3.4-fold) expression in ToF-patients. Northern blot analysis verified the expression patterns of VEGF and flk-1 in both ToF-1 and ToF-2 patients. VEGF staining in cardiomyocytes was increased in ToF-1 (1.5-fold, p < 0.05) as compared to ToF-2. Video image analysis revealed enhanced vascular density (p < 0.01) with enlarged myocyte cross sectional area (p < 0.01), but vascular wall thickness remained unchanged in ToF-1 patients as compared to age matched controls. Our data suggest that RVH is associated with profound changes in gene profile for a number of genes, where VEGF/VEGF-R system contributes to enhance, but stunted myocardial angiogenesis in patients with ToF.
法洛四联症（ToF）是一种发绀性先天性心脏病，伴有明显的右心室肥厚（RVH），伴有心肌氧和营养供应受损。因此，右心室可能表现出改变的分子表型，具有许多适应性和遗传性基因谱，这些基因谱在很大程度上是未知的。本研究的目的是研究 ToF 患者的心肌差异基因表达谱，并评估心肌血管化。对接受初次矫正手术的 ToF 患者（称为 ToF-1；n=12，平均年龄 0.5 岁）和年龄匹配的对照组（n=6）的右心室活检进行 DNA 微阵列分析，通过 Northern 杂交和 RT-PCR 进行验证。采用免疫组织化学和视频图像分析，评估血管内皮生长因子（VEGF）的表达、血管密度（α-SMA 和 CD31 染色）和心肌横截面积（Gomori 网织蛋白染色）在 ToF-1 和接受肺动脉瓣反流手术的成年患者（称为 ToF-2，n=12，平均年龄 30 岁）中的表达，并将数据与相应的年龄匹配对照组（n=6/12）进行比较。DNA 微阵列分析显示，236 种基因的表达模式发生改变，包括血管生成因子及其受体的表达增强（1.5-2.2 倍），如 VEGF、flt-1、flk-1、血管生成素-2、FGF-2、FGF-R1、PDGF-A，而 flt-4、Tie、TGF-β、TGF-β3R 的表达则降低（1.6-3.4 倍）。Northern 印迹分析验证了 ToF-1 和 ToF-2 患者中 VEGF 和 flk-1 的表达模式。与 ToF-2 相比，ToF-1 中的心肌细胞 VEGF 染色增加了 1.5 倍（p<0.05）。视频图像分析显示，血管密度增加（p<0.01），心肌横截面积增大（p<0.01），但与年龄匹配的对照组相比，ToF-1 患者的血管壁厚度保持不变。我们的数据表明，RVH 与许多基因的基因谱发生深刻变化有关，其中 VEGF/VEGF-R 系统有助于增强，但阻碍了 ToF 患者的心肌血管生成。
