[Netrin-1基因沉默对胎盘血管生成影响的体内和体外研究]

[In vivo and in vitro research study of the influence on placenta angiogenesis by gene silencing of netrin-1].

作者信息

Xie Han, Zou Li, Zhu Jian-wen, Yang Yun, Zhong Shao-ping, Wang Qian-hua

机构信息

Department of Obstetrics and Gynecology, Union Hospital, Tongji Medical College, Huazhong University of Science and Technology, Wuhan 430022, China.

出版信息

Zhonghua Fu Chan Ke Za Zhi. 2011 May;46(5):364-9.

PMID:21733374

Abstract

OBJECTIVE

To study influence on angiogenesis of placenta by gene silencing of netrin-1.

METHODS

Netrin-1 gene in human umbilical vein endothelial cells (HUVEC) and placenta of pregnant rats were silenced by RNA interference. The following methods were used in this study, including the phenytetrazoliumromide (MTT) for viability, clone formation for proliferation, transwell for migration, and tube formation for angiogenesis in vitro. The change of fetal growth was recorded. Placental microvessel density in pregnant rats was measured by immunohistochemical CD(34) staining in vivo.

RESULTS

(1) HUVEC: viability and proliferation of HUVEC were remarkably inhibited by gene silencing of netrin-1, which number of clone formation, migration cell, tube formation were from (69 ± 6)%, 86 ± 17, 37 ± 9 decreased to (46 ± 5)%, 46 ± 13 and 17 ± 5 (P < 0.05) respectively. (2) Placenta of pregnant rats: after netrin-1 gene silenced, fetal weight were decreased from (2.39 ± 0.17) g to (2.12 ± 0.10) g (P < 0.05). Placental microvessel density was decreased from (258 ± 38)/mm(2) to (197 ± 32)/mm(2) in vivo (P < 0.05).

CONCLUSIONS

Gene silencing of netrin-1 could inhibit viability, proliferation, migration, tubal formation of HUVEC and angiogenesis of placenta. Netrin-1 plays an important role in regulating angiogenesis in placenta.

摘要

目的

研究netrin-1基因沉默对胎盘血管生成的影响。

方法

采用RNA干扰技术沉默人脐静脉内皮细胞（HUVEC）和孕鼠胎盘中的netrin-1基因。本研究采用了以下方法，包括用噻唑蓝（MTT）法检测细胞活力、克隆形成实验检测细胞增殖、Transwell实验检测细胞迁移以及体外管腔形成实验检测血管生成。记录胎儿生长情况的变化。通过免疫组化CD(34)染色法在体内测量孕鼠胎盘微血管密度。

结果

（1）HUVEC：netrin-1基因沉默显著抑制了HUVEC的活力和增殖，克隆形成数、迁移细胞数、管腔形成数分别从（69±6）%、86±17、37±9降至（46±5）%、46±13和17±5（P<0.05）。（2）孕鼠胎盘：netrin-1基因沉默后，胎儿体重从（2.39±0.17）g降至（2.12±0.10）g（P<0.05）。体内胎盘微血管密度从（258±38）/mm(2)降至（197±32）/mm(2)（P<0.05）。