目的 分析gld.apoE-/-鼠外周血及骨髓内皮祖细胞(EPCs)数量及功能的变化.方法 杂交繁殖gld.apoE-/-C57BL/6雌性小鼠,采用流式细胞术分析gld+/+ apoE+/+,gld,apoE-/-,gld.apoE-/-C57BL/6雌性小鼠外周血及骨髓中CD11b-SCA-1+CD309+的EPCs的百分率；密度梯度离心和贴壁培养法分离培养小鼠骨髓EPCs,用1,1'-双十八烷基-3,3,3',3'-四甲基吲哚羰基花青高氯酸盐(DiI)标记的乙酰化低密度脂蛋白(DiI)-ac-LDL)和异硫氰酸荧光素(FITC)标记的荆豆凝集素(FITC-UEA-1)双染色标记贴壁细胞,分析EPCs吞噬功能；计数再贴壁EPCs数量以检测其黏附功能；计数血管腔样结构数量以检测其成血管功能.采用SPSS 11.5软件行t检验进行数据统计分析,GraphPad Prism 5软件作图.结果 与gld+/+ apoE+/+组[(0.039±0.005)％,P＜0.01]、gld组[(0.025±0.001)％,P＜0.05]及apoE-/-组[(0.028±0.002)％,P＜0.01]相比,gld.apoE-/-组外周血EPCs的百分率[(0.012±0.002)％]显著下降；与gld+/+ apoE+/+组[(0.42±0.05)％,P＜0.05]相比,gld.apoE-/-组骨髓EPCs的百分率[(0.12±0.01)％]显著下降；gld.apoE-/-组DiI-LDL和FITC-UEA-1双阳性细胞百分率[(59.2±2.1)％]显著低于gld+/+ apoE+/+组[(87.5±3.0)％,P＜0.01]及gld组[(84.2±6.0)％,P＜0.01]；与gld+/+ apoE+/+组[(86.0±7.3)％,P＜0.01]、gld组[(73.0±1.0)％,P＜0.01]及apoE-/-组[(65.0±4.0)％,P＜0.05]相比,gld.apoE-/-组EPCs黏附数量[(50.0±5.3)％]亦显著减少；与gld+/+apoE+/+组(12.0±1.4)相比,gld.apoE-/-组(4.0±0.3,P＜0.01)、gld组(8.0±0.5,P＜0.05)和apoE-/-组 (6.5±1.2,P＜0.01)小鼠EPCs形成血管闭合环数量明显减少,其中gld.apoE-/-组小鼠EPCs形成闭合环数量最少.结论 gld.apoE-/-C57BL/6小鼠体内EPCs数量减少,且骨髓EPCs黏附、吞噬及促血管生成功能受损,可能是狼疮伴动脉粥样硬化发生的原因之一.%Objective To detect the changes of number and function of endothelial progenitor cells (EPCs) in gld.apoE-/-C57BL/6 mice,and to investigate whether premature atherosclerosis of gld.apoE-/-C57BL/6 mice was mediated by the dysfunction of these EPCs.Methods EPCs were isolated from peripheral blood and bone marrow of four types of C57BL/6 female mice:gld+/+apoE+/+,gld,apoE-/-and gld.apoE-/-.The percentage of EPCs was analyzed by FACS as CD11b-Sca-1+CD309+ cells.The attached cells were stained with DiI labeled acetylated low-density lipoprotein (DiI-ac-LDL) and FITC-labeled Ulex Europaeus agglutinin 1 (FITC-UEA-1) double staining to determine their phagocytic function.The adherent function of EPCs was determined by calculating the number of re-cultured EPCs.The capacity of EPCs to form the cavity structure was detected by calculating the number of the formed vascular-like structure.Results The percentage of circulating EPCs was significantly decreased in the gld.apoE-/-group (0.012±0.002)％ compared to the gld+/+ apoE+/+ group [(0.039±0.005)％,P＜0.01],the gld group [(0.025±0.001)％,P＜0.05],and the apoE-/-group [(0.028±0.002)％,P＜0.01].The percentage of bone marrow derived EPCs was decreased in the gld.apoE-/-group (0.12±0.01)％ compared to the gld+/+apoE+/+ group [(0.42±0.05)％,P＜0.05].The percentage of DiI-acLDL and FITC-UEA-1 double positive cells was decreased in the gld.apoE-/-group [(59.2±2.1)％] compared to the gld+/+apoE+/+ group [(87.5±3.0)％,P＜0.01],and the gld group [(84.2±6.0)％,P＜0.01] ; the adherent function of EPCs was impaired in the gld.apoE-/-group [(50.0±5.3)％] compared to the gld+/+ apoE+/+ group [(86.0±7.3)％,P＜0.01],the gld group [(73.0±1.0)％,P＜0.01],and the apoE-/-group [(65.0±4.0)％,P＜0.05] respectively.The capacity of EPCs to form the cavity structure was decreased in the gld.apoE-/-group (4.0±0.3) compared to the gld+/+apoE+/+ group (12.0±1.4,P＜0.01).Conclusion The number of EPCs is decreased in the gld apoE-/-C57BL/6 mice,the adhesion,phagocytosis and angiogenic function of EPCs in the bone marrow are impaired,which may be one of the causes of lupus with atherosclerosis.
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