AMD 3100联合G-CSF对2型糖尿病小鼠骨髓EPCs动员的培养  

Cultivation of Bone Marrow EPCs in Type 2 Diabetic Mice by AMD3100 and G-CSF

作　　者：潘满昌 林晓莹 汪虹[1] 冷敏 PAN Man-chang;LIN Xiao-ying;WANG Hong;LENG Min(Department of Burns,the Second Affiliated Hospital of Kunming Medical University,Kunming 650000,Yunnan,China)

机构地区：[1]昆明医科大学第二附属医院烧伤科,云南昆明650000

出　　处：《医学信息》2020年第9期57-61,共5页Journal of Medical Information

基　　金：国家自然科学基金资助项目(编号:81660321)。

摘　　要：目的研究AMD3100联合G-CSF对糖尿病骨髓内皮祖细胞的培养方法和生物学功能。方法选用6周龄健康雄鼠,将10只db/db小鼠设为db/db组,10只db/+小鼠设为db/+组。采用AMD3100与CSF联合动员骨髓内皮祖细胞的方法,每只小鼠第1天注射AMD3100,后续连续注射5 d G-CSF,在第10天心脏采血制备EPCs。采用流式细胞仪检测CD34^+/CD133^+/CD309^+细胞的比例进行鉴定,鉴定成功的EPCs采用CCK-8法、Matrigel检测细胞增殖及成管功能。采用PCR检测细胞表面标志物:血管内皮生长因子(VEGF)、基质细胞衍生因子(SDF-1)、趋化因子受体4(CXCR4)基因表达情况。结果①从处理后的第3天开始,db/+组的OD450值较db/db组增高(P=0.05);②0和48 h db/+组EPCs迁移面积分别为(204088.100±219.200)μm、(144724.900±199.400)μm,大于db/db组的(200985.700±232.600)μm、(155075.300±213.100)μm(P<0.05);③db/+组管腔形成数量为(29.330±1.764)个,多于db/db组的(24.000±2.082)个(P<0.05)。结论采用AMD3100联合G-CSF动员2型糖尿病骨髓内皮祖细胞的方法成功从外周血中分离出EPCs,经体外培养发现2型糖尿病状态下EPCs的增殖,迁移和成管等功能均受损。Objective To study the culture method and biological function of AMD3100 combined with G-CSF on diabetic bone marrow endothelial progenitor cells.Methods Six-week-old healthy male rats were selected,10 db/db mice were set as db/db group,and 10 db/+mice were set as db/+group.AMD3100 and CSF were used to mobilize bone marrow endothelial progenitor cells.Each mouse was injected with AMD3100 on the first day,followed by continuous injection of G-CSF for 5 d,and blood was collected on the 10 th day to prepare EPCs.Flow cytometry was used to detect the ratio of CD34^+/CD133^+/CD309^+cells for identification.Successful EPCs were identified by CCK-8 method and Matrigel to detect cell proliferation and tube formation function.PCR was used to detect cell surface markers:vascular endothelial growth factor(VEGF),stromal cell-derived factor(SDF-1),and chemokine receptor 4(CXCR4)gene expression.Results①From the third day after treatment,the OD450 value of the db/+group was higher than that of the db/db group(P=0.05);②The migration area of EPCs in the 0 and 48 h db/+groups was(204088.100±219.200)μm,(144724.900±199.400)μm,(200985.700±232.600)μm and(155075.300±213.100)μm(P<0.05)greater than the db/db group;③The number of lumens formed in the db/+group was(29.330±1.764),which was more than(24.000±2.082)in the db/db group(P<0.05).Conclusion In this experiment,AMD3100 and G-CSF were used to mobilize type 2 diabetic bone marrow endothelial progenitor cells.EPCs were successfully isolated from peripheral blood,and the proliferation,migration and tube formation of EPCs in type 2 diabetes were impaired by in vitro culture.

关 键 词：糖尿病 内皮祖细胞 联合动员 移植 

分 类 号：R587.2[医药卫生—内分泌] R-33[医药卫生—内科学]