黄芪对2型糖尿病外周血内皮祖细胞增殖、分化的影响

Effect of Astragalus membranaceus on proliferation,differentiation of endothelial progenitor cells from peripheral blood in patients with type 2 diabetes

目的:研究黄芪对2型糖尿病(T2DM)患者外周血内皮祖细胞(endothelial progenitor cells,EPCs)增殖、分化的影响。方法:选择T2DM患者和非T2DM对照人群各20例,密度梯度离心法获取外周血单个核细胞,培养7d后,收集贴壁细胞并加入黄芪注射液(0.2,2,20,200mg/ml)干预一定时间(6h,12h,24h,48h)。激光共聚焦显微镜鉴定EPCs,MTT法、流式细胞仪检测黄芪对T2DM患者EPCs增殖、分化的影响。结果:T2DM患者外周血EPCs数量减少(P<0.01),增殖能力受损(P<0.05),EPCs数量、增殖能力与糖化血红蛋白(HbA1c)水平和糖尿病病程年呈负相关(P<0.01);T2DM患者外周血EPCs CD14+、CD64+细胞百分比明显高于对照组,而vWF+细胞百分比明显低于对照组(P<0.01)。黄芪对T2DM患者外周血EPCs具有明显的促增殖及促进其向内皮细胞系分化的作用。结论:T2DM患者外周血EPCs数量减少、增殖及分化能力受损;黄芪对T2DM患者外周血EPCs增殖及向内皮细胞系分化有显著促进作用。

Objective： To study the effects of Astragalus membranaceus on the proliferation, differentiation of endothelial progenitor cells（EPCs） from peripheral blood in patients with type 2 diabetes. Methods： Total mononuclear cells were isolated from patients with type 2 diabetes （n=20） and age-matched control subjects （n=20）by Ficoll density gradient centrifugation, and then the cells were plated on fibronectin coated culture dishes. After 7 days of culture, attached cells were stimulated with Astragalus memhranaceus in a series of concentrations （0.2, 2, 20and200mg/ml） for different durations （6, 12, 24, and 48h）. EPCs were characterized as adherent cells double positive for DiLDL-uptake and lectin binding by direct fluorescent staining under a laser scanning confocal microscope. The effect of Astragalus membranaceus on EPCs proliferation, differentiation was measured by MTT assay and flow cytometry. Results： The number and proliferation of EPCs was significantly reduced in patients with type 2 diabetes as compared with that in control subjects（P〈0.01,P〈0.05）, and it was inversely correlated with HbAlc levels and the course of diabetes （P〈0.01）; the relative proportion of diabetic EPCs expressing CD14 and CD64 was increased and expressing the endothelial marker yon Willebrand factor was reduced as compared with that in control subjects（P〈0.01）. Astragalus membranaceus could markedly promote the proliferation and the differentiation toward the endothelial cell lineage of EPCs in T2DM in a time- and dose-dependent manner. Conclusion： Astragalus membranaceus could markedly promote the proliferation and the differentiation toward the endothelial cell lineage of EPCs in T2DM.