摘要
目的观察外源性Wnt-5a对小鼠内皮祖细胞(EPC)增殖﹑迁移和成管效应的影响,了解Wnt-5a在血管发生中的作用。方法密度梯度离心法获取小鼠骨髓源性单个核细胞,应用EGM-2培养基诱导培养为EPC。在加入外源性Wnt-5a细胞因子的条件下,采用Ki-67细胞免疫荧光染色法检测细胞增殖情况,采用Boyden Chamber Transwell法检测其对EPC迁移能力的影响,用Matrigel基质胶成管实验检测其对EPC成管效应的影响。结果实验组Ki-67阳性细胞百分比为(16.7±0.3)%,与对照组(12.5±0.2)%比较差异有统计学意义(P<0.05)。培养48 h后对照组每视野迁徙细胞数为(87.7±11.3)个,实验组为(112.4±12.2)个,2组比较差异有统计学意义。Matrige1基质胶成管实验亦提示外源性Wnt-5a对EPC的成管效应有促进作用。结论外源性Wnt-5a能够促进小鼠EPC的血管发生,提示非经典Wnt信号通路可能在EPC血管发生过程中发挥重要作用。
Objective To investigate the effect of exogenous Wnt-5a on proliferation,migration and tube formation of mouse endothelial progenitor cell（EPC）,so as to realize the role of Wnt-5a in angiogenesis.Methods Bone marrow-derived mononuclear cells isolated by density gradient centrifugation were cultured in EGM-2 medium,and then were induced into EPC.After treatment of exogenous Wnt-5a,the cell proliferation ability was detected by Ki-67 immunofluorescence staining method,and EPC＇s migration ability and tube formation ability were detected with the method of Boyden Chamber Transwell and Matrigel tube formation,respectively.Results The number of Ki-67 positive cells in experimental group was（16.7±0.3）% per high power field,which was statistically significant compared with control group（12.5±0.2）%.After culture for 48 h in Boyden Chamber,the number of migrated cells in control group was（87.7±11.3） per field,while the number in experimental group was（112.4±12.2） per field,and the difference was statistically significant.Matrigel tube formation test also indicated Wnt-5a could enhance EPC＇s tube formation ability.Conclusion Exogenous Wnt-5a can enhance vasculogenesis in which EPC participates,which indicates noncanonical Wnt pathway may play an important role in the process of vasculogenensis.
出处
《中国医科大学学报》
CAS
CSCD
北大核心
2012年第3期228-230,233,共4页
Journal of China Medical University
基金
辽宁省教育厅创新团队计划(2008T201)
辽宁省教育厅优秀人才支持计划(2008RC53)
