内皮型一氧化氮合酶基因体外转染对犬内皮祖细胞功能的影响  

Effect of heNOS gene transfection on functions of canine bone marrow-derived endothelial progenitor cell

作　　者：郭连瑞[1] 张建[1] 宋礼坡[1] 王国栋[1] 谷涌泉[1] 张淑文[1] 李建新[1] 汪忠镐[1] 

机构地区：[1]首都医科大学宣武医院血管外科,北京100053

出　　处：《中国普通外科杂志》2011年第12期1331-1335,共5页China Journal of General Surgery

基　　金：国家自然科学基金资助项目(30471708)

摘　　要：目的探讨人内皮型一氧化氮合酶基因(heNOS)转染对犬骨髓源内皮祖细胞(EPC)功能的影响。方法用携带heNOS基因的5型腺病毒作为载体(Ad5-heNOS),对体外定向培养扩增的骨髓源犬EPC进行转染,以未转染的犬EPC作为对照。用酶联免疫吸附试验(ELISA)和硝酸还原酶法检测转染后的EPC中heNOS蛋白的表达和NO的产量,并检测转染后EPC的增殖、黏附、迁移、抗衰老和成血管能力等功能。结果 Ad5-heNOS转染48 h后,转染组EPC的eNOS蛋白表达量和NO产量均明显高于未转染组[(2091.67±172.489)pg/mL vs.(158.00±30.914)pg/mL;(49.5±5.16)μmol/Lvs.(39.7±7.24)μmol/L](均P<0.01);转染组EPC的细胞增殖数、黏附数、迁移数均明显高于未转染组(0.52±0.03 vs.0.31±0.02;28.00±1.41 vs.11.83±1.45;109.67±6.95 vs.72.67±6.29)(均P<0.01),而衰老细胞百分数明显低于未转染组(0.22±0.02 vs.0.32±0.01)(P<0.01);转染heNOS基因的EPC在体外基质胶中培养可形成血管样结构。结论 heNOS基因转染能促进犬EPC的增殖并增强其功能。Objective To investigate the effect of human eNOS（heNOS） gene transfection on the functions of canine bone marrow-derived endothelial progenitor cells（EPC）.Methods Canine EPCs were transfected with the recombinant adenovirus vector type 5 carrying heNOS gene（Ad5-heNOS） after directional culture and expansion ex vivo,and the untransfected canine EPCs were used as control.After transfection,the heNOS protein expression and NO production of EPCs were measured by enzyme linked immunosorbent assay（ELISA） and nitrate reductase assay,respectively.The EPCs functions including proliferation,adhesion,migration and anti-senescence as well as angiogenesis ability were also determined.Results Forty-eight hours after transfection with Ad5-heNOS,the heNOS protein expression and NO production of transfected EPCs were significantly higher than those of the untransfected EPCs [（2091.67±172.489） pg/mL vs.（158.00±30.914） pg/mL;（49.5±5.16） μmol/L vs.（39.7±7.24） μmol/L]（both P0.01）.The cell number of proliferation,adhesion and migration of the transfected EPCs were all significantly higher than those of the untransfected EPCs（0.52±0.03 vs.0.31±0.02;28.00±1.41 vs.11.83±1.45;109.67±6.95 vs.72.67±6.29）（all P0.01）,while the percentage of senescent cells of the transfected EPCs was significantly lower than that of the untransfected EPCs（0.22±0.02 vs.0.32±0.01）（P0.01）.The transfected EPCs assembled into primitive vascular tube-like structures when plated in Matrigel ex vivo.Conclusions heNOS gene transfection can promote the proliferative ability of canine EPC and also enhance its functions.

关 键 词：内皮祖细胞 内皮型一氧化氮合酶 转染 血管新生 犬 

分 类 号：R329.4[医药卫生—人体解剖和组织胚胎学]