外周血两种不同类型内皮祖细胞与颈动脉狭窄的关系

The Relationship Study Between Two Types of Endothelial Progenitor Cells in Peripheral Blood and the Cerebral Vascular Stenosis

目的探讨外周血中早期内皮祖细胞和晚期内皮祖细胞数量和功能变化与颈动脉狭窄程度之间的关系。方法入组对象60例,根据全脑血管造影术分成颈动脉狭窄组40例(其中轻度狭窄组20例,中重度狭窄组20例),对照组20例。在全脑血管造影术中抽取患者股动脉血,采用密度梯度离心法分离外周血单个核细胞,分别培养至7天和21天鉴定为早期内皮祖细胞、晚期内皮祖细胞,计数细胞集落数量并分别用MTT比色法、改良的Boyden小室法和HFN培养板测定早期内皮祖细胞、晚期内皮祖细胞的增值、迁移、黏附功能。结果颈动脉狭窄组患者吸烟、高血压、甘油三酯、低密度脂蛋白明显高于对照组,高密度脂蛋白低于对照组(P<0.05);而年龄、性别、血糖、总胆固醇与对照组比较,差异无统计学意义(P>0.05)。与对照组比较,颈动脉狭窄组外周血早期内皮祖细胞、晚期内皮祖细胞数量及功能均下降(P<0.05),晚期内皮祖细胞数量与功能改变与颈动脉狭窄程度呈负相关(P<0.05)。结论颈动脉狭窄患者外周血晚期内皮祖细胞数量减少,功能受损,其数量与功能变化与颈动脉狭窄程度呈负相关。对于颈动脉狭窄患者,检测晚期内皮祖细胞数量及功能可间接预测颈动脉狭窄严重程度。

Aim To discusse the relationship between the number and function of early and late endothelial progenitor cell （EPC） in peripheral blood and the carotid stenosis （CS）. Methods 60 cases were included into the group of objects, and divided into group of dO patients with carotid artery stenosis （mild stenosis in 20 cases, severe stenosis in 20 cases）, normal control group of 20 cases. The blood was extracted from the patients＇ femoral artery in the procedure of cerebrovascular digital subtraction angiography, mononuclear cells were isolated from peripheral blood by density-gradient centrifugation and were cultured for 7 days and 21 days to be identified as early endothelial progenitor cells and late endothelial progenitor cells, colony counting the number of early EPC and late EPC. Proliferation and migration were determined by MTT assay and modified Boyden. The two types of EPCs adhesion assay were performed by replating those on fibronectin-coated dishes, then adherent cells were counted. Results Smoking, high blood pressure and triglycerides, low density lipoprotein in carotid stenosis groups were significantly higher than those in the control group, high-density lipoprotein cholesterol （HDLC） was lower than those of the control group （P 〈 0. 05） ; the difference of age, sex, blood sugar, total cholesterol compared with controls, was not statistically significant （P 〉 0. 05）. Compared with controls, the number and function of early EPC, late EPC in carotid stenosis group was decreased, the number and function changes of late EPC were negatively correlated with the degree of cerebral stenosis （P 〈 0. 05）. Conclusions Decreased late EPC colonies amount and functional impairment can be observed in patients with carotid artery stenosis. The number and function changes of late EPC were negatively correlated with the degree of cerebral vascular stenosis. Detection of late EPC number and function can be indirect prediction of carotid stenosis severity.