复方丹参注射液对内皮祖细胞数量和功能的影响

Effects of Compound Salvia injection on number and activity of endothelial progenitor cells

目的:观察复方丹参注射液对内皮祖细胞(endothelial progenitor cells,EPCs)数量和功能的影响。方法:采用密度梯度离心法分离人脐血单个核细胞,接种到包被有人纤维连接蛋白的培养皿,分为对照组(未加细胞因子)、VEGF组(加血管内皮生长因子10 ng.mL-1)、复方丹参50组、复方丹参10组及复方丹参2组(分别加入复方丹参注射液50,10,2μg.mL-1),培养3 d后,倒置显微镜下观察集落形成,采用流式细胞术测定各组EPCs数,采用黏附能力测定试验测定黏附能力。结果:VEGF组、复方丹参10组及复方丹参2组的EPCs数显著高于对照组(分别P<0.01,P<0.05,P<0.01),且复方丹参2组显著高于复方丹参10组及VEGF组(均P<0.01);复方丹参50组的EPCs数显著低于对照组(P<0.01)。复方丹参10组及复方丹参2组的集落数及黏附细胞数显著高于对照组,而复方丹参50组则显著低于对照组。结论:低剂量复方丹参显著提高培养EPCs数量和功能,高剂量复方丹参则反之。

Objective： To investigate the effects of Compound Salvia injecfion（CSI） on the number and activity of endothelial progenitor cells（EPCs）. Method： Mononuelear fraction of human umbilical cord blood was obtained by density gradient centrifugation and plated on fibronectin coated culture dishes. Cells were divided in to five groups： group control, group VEGF, group CSI 50, group CSI 10 and group CSI 2 （supplemented with none cytokine, VEGF 10 ng·mL^-1, CSI 50, 10, 2 μg· mL^-1, respectively）. After six days in culture, cell clusters were viewed with an inverted microscope, fluorescence-activated cell sorting（FACS） analysis of PE-CD34 and FITC-VE-Cadherin was performed to detect number of EPCs, adhesion assay was performed by replating cells on fibronectin coated dishes, and then counting adherent cells. Result： Numbers of EPCs of group VEGF, group CSI 10 and group CSI 2 were signitlcanfly increased as compared with those of group control（ P 〈 0.01, P 〈 0.05, P 〈 0.01, respectively）, and numbers of EPCs of group CSI 2 were more than those of group CSI 10 and group V（ P 〈 0.01）. Compared with group control, number of EPCs of group CSI 50 was significantly decreased（ P 〈 0.01）. Compared with group control, numbers of clusters and adhesive EPCs of group CSI 10 and group CSI 2 were significantly increased, while those of group CSI 50 were significantly decreased. Conclution： Low concentration CSI can significantly promote EPCs augmentation and enhance its functional activity, while high concentration CSI significantly restrains it.