脑钠肽对外周血内皮祖细胞增殖、迁移与黏附能力的影响

Effects of brain natriuretic peptide on proliferation,migration and adhesion of endothelial progenitor cells from peripheral blood

目的研究脑钠肽对外周血内皮祖细胞数量及增殖、迁移、黏附能力的影响。方法选取30例健康成人,取其外周静脉血,采用密度梯度离心法分离外周血获得单个核细胞,将其接种在人纤维连接蛋白包被的培养板上,经血管内皮生长因子(Vascular endothelial growth factor,VEGF)诱导培养,7d后获得贴壁细胞,荧光显微镜鉴定FITC-UEA-I和Dil-acLDL双染色为正在分化的内皮祖细胞(endothelial progenitor cells,EPCs)。收集EPCs随机分成6组,分别为对照组和不同浓度脑钠肽干预组(0.05μg/ml、0.10μg/ml、0.15μg/ml、0.20μg/ml、0.25μg/ml),均培养24h后采用MTT比色法测定内皮祖细胞(EPCs)的增殖能力,改良的Boyden小室法测定EPCs的迁移能力,黏附实验测定内皮祖细胞的黏附能力。结果不同浓度的重组人脑钠肽干预组均显著增加EPCs的增殖及迁移能力,分别与两者呈一定的量效关系,对黏附能力无明显影响。结论脑钠肽能促进外周血内皮祖细胞增殖、迁移能力,对内皮祖细胞的黏附能力无明显影响。

Objective To study the effects of brain natriuretic peptide （BNP） on proliferation, migration, and adhesion of en- dothelial progenitor cells （EPCs） from peripheral blood. Methods Mononuclear cells were isolated from peripheral blood of 30 healthy aduh venous blood by density gradient centrifugation,and the ceils were plated on fibronectin-coated culture plates and induced by cultured vascular endothelial growth factor, after 7 days adherent cells were obtained. FITC-UEA-I and Dil-acLDL double staining of differentiating endothelial progenitor cells were identified by fluorescence microscopy. The endothelial progenitor cells were collected and were randomly divided into six groups： the control group and different concentrations of brain natriuretic peptide intervention group（0.051~g/ml, 0.10lxg/ml, 0.151xg/ml, 0.201xg/ml, 0.25txg/ml）. After 24 hours of being cultured, progeni- tor cell proliferation was measured by MTY assay, migration of endothelial progenitor cell by modified Boyden chamber assay, ad- hesion of endothelial progenitor cells by adhered experimental determination. Results Different concentrations of recombinant hu- man brain natriuretic peptide in the intervention group significantly increased proliferation and migration of EPCs, and showed a dose-effect relationship between them, no significant effect on adhesion. Conclusion Brain natriuretic peptide could promote en- dothelial progenitor cell proliferation, migration, had no significant effect on adhesion endothelial progenitor cells.