[1]潘满昌,林晓莹,汪 虹,等.AMD3100联合G-CSF对2型糖尿病小鼠骨髓EPCs动员的培养[J].医学信息,2020,33(09):57-61.[doi:10.3969/j.issn.1006-1959.2020.09.018]
　PAN Man-chang,LIN Xiao-ying,WANG Hong,et al.Cultivation of Bone Marrow EPCs in Type 2 Diabetic Mice by AMD3100 and G-CSF[J].Medical Information,2020,33(09):57-61.[doi:10.3969/j.issn.1006-1959.2020.09.018]
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AMD3100联合G-CSF对2型糖尿病小鼠骨髓EPCs动员的培养()
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医学信息[ISSN:1006-1959/CN:61-1278/R]

卷:
33卷
期数:
2020年09期
页码:
57-61
栏目:
论著
出版日期:
2020-05-01

文章信息/Info

Title:
Cultivation of Bone Marrow EPCs in Type 2 Diabetic Mice by AMD3100 and G-CSF
文章编号:
1006-1959（2020）09-0057-06
作者:
潘满昌; 林晓莹; 汪 虹; 等
（昆明医科大学第二附属医院烧伤科，云南 昆明 650000）
Author(s):
PAN Man-chang; LIN Xiao-ying; WANG Hong; et al
(Department of Burns,the Second Affiliated Hospital of Kunming Medical University,Kunming 650000, Yunnan,China)
关键词:
糖尿病; 内皮祖细胞; 联合动员; 移植
Keywords:
Diabetes; Endothelial progenitor cells; Joint mobilization; Transplantation
分类号:
R587.2；R-33
DOI:
10.3969/j.issn.1006-1959.2020.09.018
文献标志码:
A
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。
Abstract:
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 10th 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.Results ①From 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.

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更新日期/Last Update: 2020-05-01