血液透析患者内皮祖细胞的变化及其与心血管危险因素的关系

Changes of Endothelial Progenitor Cells and the Correlation with the Risk Factors of Coronary Heart Disease in Chronic Renal Failure Patients Subject to Long-term Hemodialysis

目的检测血液透析患者内皮祖细胞数量(EPCs)和功能的改变,并探讨其与心血管危险因素的关系。方法采用密度梯度离心法分离培养慢性肾功能衰竭长期血液透析患者和健康组外周血单个核细胞,将其接种在人纤维连接蛋白包被培养板,7d后取贴壁细胞进行Di-LDL和FITC-UEA-I双染色,并通过流式细胞仪检测其表面标志CD34、CD133、KDR,以鉴定EPCs。采用改良的Boy-den小室、黏附功能检测评价其迁移和黏附能力,并与性别、高血压、糖尿病、吸烟及高脂血症等心血管危险因素和Framingham危险评分进行统计学分析。结果长期血液透析患者EPCs的数量和迁移、黏附功能都低于正常健康者,差异有统计学意义(P<0.05);有糖尿病的患者EPCs的数量和黏附、迁移功能明显降低(P<0.05),有高血压的患者EPCs的数量和迁移能力明显降低(P<0.05),而性别、血脂和吸烟对EPCS有一定的影响但差异无统计学意义;Framingham危险评分与患者EPCs的数量和迁移、黏附功能分别呈明显负相关(r=-0.623,-0.658,-0.625,P<0.05)。结论长期血液透析患者的EPCs数量和功能降低,存在血管新生和内皮修复的缺陷,增加了患冠心病的风险。

Objective To investigate the number and activity changes of endothelial progenitor cells （EPCs） in chronic renal failure patients subject to long-term hemodialysis, and the correlation with the risk factors of coronary heart disease. Methods Total mononuclear cells（MNCs） were isolated from peripheral blood of patients with chronic renal failure on maintenance hemodialysis and a normal control group by Ficoll density gradient centrifugation and then were planted on humanfibronectin-coated dishes. After 7 days of culture, EPCs were characterized as adherent cells by double staining FITC-UEA-I binding and DI-LDL, and were further documented by demonstrating the expression of CD34, CD133 and KDR with flow cytometry. EPCs migration was determined with modified Boyden chamber assay. EPCs adhesive assay was performed by replanting EPCs on humanfibronectin-coated plates and then counting the adherent cells. The relationship between the EPCs＇ number and activity and the risk factors of CHD（ such as gender, hypertension, hypercholesterolemia, diabetes, smoking） and Framingham Risk Score were assessed. Results Number of EPCs and migration and adhesive capacity were significantly lower（ P 〈 0.05）; diabetes was associated with significantly lower EPCs＇ number and migration, adhere activity, and hypertension with EPCs＇ number and migration（P 〈 0.05）, whereas a minor but nonsignificant reduction of EPCs levels and activity were detected in the presence of gender, hypercholesterolemia, and smoking; Framingham Risk Score was significantly correlated with a reduction of EPCs＇ level and activity（ r = - 0. 623, - 0. 658, - 0. 625, P 〈 0.05）. Conclusion The number and activity of EPCs were decreased in patients with chronic renal failure on maintenance hemodialysis, neovascularization and repairment of damaged endothelium were impaired in these patients, which were related to the increased risk of coronary heart disease.