基础医学与临床 ›› 2014, Vol. 34 ›› Issue (10): 1405-1409.

• 研究论文 • 上一篇    下一篇

VEGF通过上调CCN2促HUVEC迁移和血管生成

曹玉净,吕秋霞   

  1. 河南省中医院
  • 收稿日期:2013-12-27 修回日期:2014-04-21 出版日期:2014-10-05 发布日期:2014-09-25
  • 通讯作者: 吕秋霞 E-mail:changliuxian@gmail.com

VEGF Promotes HUVEC Migration and Angiogenesis via up-Regulation of CCN2

  • Received:2013-12-27 Revised:2014-04-21 Online:2014-10-05 Published:2014-09-25

摘要: 目的 探讨血管内皮生长因子(VEGF)诱导的成骨细胞中结缔组织生长因子(CTGF/CCN2)对人脐静脉血管内皮细胞(HUVECs)的影响。方法 用Real time PCR法及ELISA法检测VEGF诱导成骨细胞(OSE)中CCN2含量;制备成骨细胞(OSE)上清液;将细胞分为control组、OSE组和VEGF-OSE组(n=3)。用小干扰RNA(siRNA)转染法抑制成骨细胞中CCN2的表达;Transwell法检测内皮细胞迁移;Matrigel实验检测管样结构形成能力。结果 VEGF呈时间和剂量依赖性上调成骨细胞中CCN2 mRNA和蛋白的表达;CCN2可促进内皮细胞的迁移和管样结构形成(P<0.05),当CCN2被siRNA基因沉默或者加入CCN2抗体后,CCN2对内皮细胞迁移和管样结构形成的促进作用均受到明显抑制(P<0.05)。结论 VEGF通过上调成骨细胞中CCN2的表达,促内皮细胞(HUVECs)的迁移和血管生成。

关键词: VEGF, CCN2, 成骨细胞, 人脐静脉血管内皮细胞, 血管生成

Abstract: Objective: To explore the effect of VEGF-induced up-regulation of CCN2 in osteoblasts mediates biological behavior in human umbilical endothelial cells. Methods: Expression of CCN2-mRNA and CCN2-protein were examined in osteoblates in the presence of VEGF by Real time PCR and ELISA. Generation of osteoblast-conditioned supernatant. HUVECs were divided into three groups: Control group, OSE group, VEGF-OSE group (n=3). The chemically synthesized CCN2-siRNA was transfected into osteoblasts. The ability of cell migration was analyzed by the Transwell assay. The capability of cell to form capillary-like tubes in vitro was assessed on Matrigel assay. Results: VEGF increases the expression of CCN2 mRNA and protein in osteoblasts in a time- and dose-dependent manner. CCN2 promotes HUVEC migration and capillary tube-forming in vitro(P<0.05). These effects were prevented by an antibody against CCN2 or by small interfering RNA-mediated knockdown of osteoblast CCN2 (P<0.05). Conclusion: VEGF-induced up-regulation of CCN2 in osteoblasts mediates the ability of migration and angiogenesis in HUVEC.

Key words: VEGF, CCN2, Osteoblasts, Human umbilical endothelial cells, Angiogenesis