机构地区:[1]中南大学湘雅二医院胸外科,湖南长沙410011 [2]湖南省现代优生技术重点实验室,湖南长沙410008
出 处:《中国当代儿科杂志》2009年第4期267-272,共6页Chinese Journal of Contemporary Pediatrics
基 金:湖南省自然科学基金(06JJ50031)
摘 要:目的探讨紫绀型先天性心脏病患儿血清血管内皮生长因子(VEGF)和基质细胞衍生因子(SDF-1)的水平与循环内皮祖细胞(EPCs)数量及功能变化的关系。方法采用ELISA法检测15例法洛四联症患儿(紫绀组)外周血清SDF-1和VEGF的水平;采用密度梯度离心法分离EPCs体外扩增培养;做免疫荧光鉴定及细胞计数。采用MTT比色法、改良的Boyden小室和黏附能力测定实验,观察EPCs的增殖能力、迁移能力及黏附能力。用回归分析法分析VEGF和SDF-1水平与EPCs数量及功能的相关性。单纯室间隔缺损患儿15例为对照组。结果紫绀组与对照组相比,VEGF水平增高(201.42±44.74 ng/L vs 113.56±35.62 ng/L);SDF-1水平增高(3.45±1.07 ng/L vs 1.05±0.99 ng/L,P<0.05)。血清VEGF与SDF-1水平呈正相关(r=0.675,P<0.01)。与对照组相比,紫绀组PEPCs数目明显增多(72.2±9.73)/200倍视野vs(51.2±3.83)/200倍视野,P<0.01;EPCs功能明显增强,其增殖能力、迁移能力及黏附能力明显强于对照组。VEGF,SDF-1的血清水平与EPCs数量及功能变化表现出明显的一致性,相关系数分别为0.8395,0.5491,0.6376和0.7392。结论紫绀型先天性心脏病外周血EPCs数量及功能较非紫绀组明显增加,同时VEGF和SDF-1血清水平也明显增加,上述变化存在相关性,可能共同影响着患者的一些病理、生理变化。Objective To examine the number and function of circulating endothelial progenitor cells ( EPCs ) in children with cyanotic congenital heart diseases (CHD) and study their correlation with serum levels of vascular endothelial growth factor (VEGF) and stromal cell derived factor-1 ( SDF-1 ). Methods Fifteen children with tetralogy of Fallot (cyanotic group) and 15 age- and sex-matched children with ventricular septal defect (control group) were enrolled. Serum levels of VEGF and SDF-1 were measured using ELISA. Mononuclear cells were isolated from peripheral blood by Ficoll density gradient centrifugation and cultured in vitro. EPCs were identified by immunofluorescence and were counted under a microscope. Modified Boyden chamber assay and the MTT assay were used to measure the migration and proliferation capacities of EPCs. EPCs adhesion ability assay was performed by replating cells on fibronectin-coated dishes, and then adherent cells were counted. The correlations of serum levels of VEGF and SDF-1 with the number and function of circulating EPCs were assessed by linear regression analysis. Results Serum levels of VEGF ( 201.42 ± 44.74 ng/L vs 113.56 ± 35.62 ng/L; P 〈 0.05) and SDF-1 (3.45 ±1.07 ng/L vs 1.05 ±0.99 ng/L; P 〈 0.05 ) in the cyanotic group were higher than those in the control group. There was a positive correlation between serum levels of VEGF and SDF-1 (r = 0.675, P 〈 0.01 ). The number of EPCs ( x 200 field) in the cyanotic group significantly increased compared with that of the control group (72. 2 ± 9. 73 vs 51. 2 ± 3. 83; P 〈 0.01 ). The functional activities of EPCs, including proliferation, migration and adhesion capacities, were augmented in the cyanotic group compared with those in the control group. The increased number and function of EPCs and the increased serum levels of VEGF and SDF-1 were consistent in the cyanotic group, with a correlation coefficient of 0. 8395,0.5491,0.6376 and 0. 7392 respectively. Conclusions The number
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