摘要
目的建立基于人脐静脉内皮细胞(HUVEC)、纤维蛋白基质及微载体的体外三维新生血管模型,在此模型的基础上,研究CD105在血管新生过程中的表达变化及其作用。方法分离、纯化并培养HUVEC;建立三维新生血管模型:将HUVEC包被于微载体(Cytodex-3)上,然后将该包被有HUVEC的微载体包埋于纤维蛋白凝胶基质中,在培养体系中内皮细胞生长因子的作用下完成HUVEC的出芽、分支、连接成网等过程。通过逆转录-聚合酶链反应(RT-PCR)检测血管新生不同时期CD105的表达情况,并利用反义寡核苷酸技术干预CD105的表达,观察抑制CD105表达后HUVEC与血管新生过程的表形变化。结果包埋于三维纤维蛋白基质中的HUVEC出现出芽、分支、连接成网等典型的血管新生过程;在此过程中,CD105的表达在出芽早期及出芽分支期明显增强,而在出芽前及成网后的相对稳定期则呈不表达或弱表达状态。利用反义寡核苷酸抑制CD105表达后,HUVEC的生长及新生血管模型中的出芽、分支过程均受到明显抑制。结论CD105在血管新生的不同阶段表达水平不同,在出芽、分支期明显过表达;CD105参与了血管新生过程,抑制CD105的表达可以有效地抑制血管新生。
Objective To develop an in vitro three-dimensional angiogenesis system and analyze the expression and function of CD105 in angiogenesis.Methods After primary human umbilical vein endothelial cells（HUVEC） were purified and cultured,the microcarriers were coated with HUVEC and then embedded and cultured into fibrin gel.The angiogenesis process of HUVEC on the microcarriers was formed.The expression of CD105 during this process was detected by reverse transcription polymerase chain reaction（RT-PCR）.Antisense oligodeoxynucleotide（ASODN） was used to inhibit the expression of CD105 and the changes of the angiogenesis process were analyzed quantitatively.Results HUVEC on the microcarriers which were embedded into the fibrin gel,occurred the angiogenesis process of sprouts,branches and capillary networks with lumina.During this process,CD105 was over expressed in the periods of forming sprouts and branches,and depressed in the relatively steady periods including the periods before forming sprouts and after forming capillary networks.While the expression of CD105 was inhibited by ASODN,the angiogenesis process was significantly inhibited.Conclusions The expression of CD105 is varied within the angiogenesis process,over expressing during the sprouts and branches forming periods.Inhibiting the expression of CD105 could efficiently inhibit angiogenesis.
出处
《中国普外基础与临床杂志》
CAS
2009年第10期824-829,共6页
Chinese Journal of Bases and Clinics In General Surgery
关键词
新生血管
人脐静脉内皮细胞
CD105
三维新生血管模型
微载体
Angiogenesis
Human umbilical vein endothelial cell
CD105
Three-dimensional angiogenesis system
Microcarrier
