老年2型糖尿病患者外向生长内皮细胞功能的变化

目的 探讨老年2型糖尿病患者外向生长内皮细胞(outgrowth endothelial cells,OEC)数量及功能的变化. 方法选取老年糖尿病患者(糖尿病组)30例及健康对照者(对照组)30例,受检者分别抽取外周血 20 ml,密度梯度离心法分离单个核细胞后于体外培养,并通过流式细胞仪测定CD14、CD34、CD45、CD105、CD133、血管内皮生长因子受体2(KDR)、血管性血友病因子(vWF)的表达情况;激光共聚焦鉴定OEC吞噬乙酰化低密度脂蛋白(ac-LDL)及结合荆豆凝集素(UEA-1)的能力;计数培养16 d形成的OEC集落数,取P1代细胞,分别采用噻唑兰(MTT)法、黏附能力测定实验和硝酸还原酶法测定两组OEC的增殖能力、黏附功能以及一氧化氮(NO)、一氧化氮合酶(NOS)的含量. 结果糖尿病组和对照组的OEC流式鉴定结果均为CD14和CD45阴性,CD133、CD34、CD105、KDR、vWF阳性,两组培养的OEC均町以吞噬ac-LDL并结合UEA-1;糖尿病组OEC集落数显著小于对照组[(2.20±1.13)CFU对(3.63±1.29)CFU,T=4.565,P<0.01];糖尿病组细胞增殖力[(0.307±0.031)A],较对照组[(0.484±0.070)A]减弱(t=11.519,P<0.01);糖尿病组细胞黏附功能[(37.74±8.37)个/HP],较对照组[(51.66±11.39)个/HP]减弱(t=5.513,P<0.01);糖尿病组合成NO[(273.40±49.85)μmol/L]和NOS能力((38.05±8.68)U/m1],均较对照组((362.23±49.88)μmol/L和(51.61±10.29)U/ml]显著降低(t=6.894,5.535,均P< 0.01). 结论老年糖尿病患者外向生长内皮细胞数量和功能均下降.

Type 2 diabetes mellitus impair the function of outgrowth endothelial cells in the aged

Objective To explore the quantity and function of outgrowth endothelial cells (OECs) in the aged with type 2 diabetes mellitus. Methods The 30 aged diabetics were as diabetes group, while 30 healthy people were as control group. And 20 ml peripheral blood were collected from every subjects, then the mononuclear cells were isolated by Ficoll density gradient centrifugation. The expressions of CD14, CD34, CD45, CD105, CD133, KDR(vascular endothelial growth factor receptor 2) and von Willebrand factor (vWF) were measured. And OECs were assessed by phagocytizing acetylated low density lipoprotein(ac-LDL) and binding Ulex europaeus agglutinin KUEA-1). The proliferation of OECs was assayed by 5-diphenyl tetrazolium bromide (MTT) assay. The adhesion assay was performed by replating cells on fibroncctin-coatcd culture dishes. NO and NOS were assayed by nitrate reductase. Results The number of OECs was lower in diabetes group than in control group. In addition, the diabetes group versus control group showed the impaired OECs functions in proliferationC(0.307 ±0.031)A vs. (0. 484 ±0.070)A, P < 0.01], adhesionC(37. 74 ±8.37)/HP vs. (51. 66±11. 39)/HP, P< 0. 01], NO production[( 273. 40±49. 85) μmol/L vs. (362. 23±49. 88) μmol/L, P < 0. 01] and NOS production C( 38.05 ±8.68 ) U/ml vs. ( 51. 61±10. 29 ) U/ml, F < 0. 01]. Conclusions Type 2 diabetes may impair the quantity and function of OECs in the aged.