复方丹参注射液对内皮祖细胞数量和功能的影响

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。

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

OBJECTIVE: To investigate the effects of Compound Salvia injection (CSI) on the number and activity of endothelial progenitor cells (EPCs). METHOD: Mononuclear 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 x mL(-1), CSI 50, 10, 2 microg x 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 significantly 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. CONCLUSION: Low concentration CSI can significantly promote EPCs augmentation and enhance its functional activity, while high concentration CSI significantly restrains it.