文献类型：期刊文献

中文题名：低氧下血管内皮细胞生长因子转染人骨髓间充质干细胞向血管内皮样细胞的分化

英文题名：Vascular endothelial growth factor transfection induces human bone marrow mesenchymal stem cells differentiating into endothelial-like cells under hypoxia

作者：胡继宏[1];贾佳[2];路娟[1];王秋萍[1];赵静苗[1];靳利梅[1];李金娟[1]

第一作者：胡继宏

机构：[1]甘肃中医药大学,甘肃省兰州市730000;[2]西安海棠职业学院,陕西省西安市713000

第一机构：甘肃中医药大学

年份：2017

卷号：21

期号：9

起止页码：1352

中文期刊名：中国组织工程研究

外文期刊名：Chinese Journal of Tissue Engineering Research

收录：CSTPCD;;Scopus;北大核心:【北大核心2014】；

基金：甘肃省高校基本科研业务费项目(2012-3)~~

语种：中文

中文关键词：干细胞;间质干细胞;血管内皮生长因子类;内皮细胞;组织工程;骨髓干细胞;血管内皮细胞生长因子;骨髓间充质干细胞;血管内皮细胞;内皮型一氧化氮合成酶;内皮素;前列环素

外文关键词：Stem Cells; Mesenchymal Stem Cells; Vascular Endothelial Growth Factors; Endothelial Cells; Tissue Engineering

摘要：背景:已有研究发现血管内皮细胞生长因子可诱导骨髓间充质干细胞分化为内皮细胞,但在机体损伤的低氧环境下,血管内皮细胞生长因子基因是否可促进骨髓间充质干细胞定向诱导分化为血管内皮样细胞?目的:观察低氧环境下,经血管内皮细胞生长因子诱导的人骨髓间充质干细胞是否能够向血管内皮样细胞分化。方法:在含体积分数5%O2环境中,将第3代人骨髓间充质干细胞分组培养,对照组以常规培养基培养,实验组以含血管内皮细胞生长因子载体的腺病毒诱导液培养。培养2周后,进行形态学观察及表面相关分子检测,ELISA检测细胞培养上清液中血管内皮细胞生长因子及内皮型一氧化氮合成酶因子水平,RT-PCR和Western blotting检测内皮素和前列环素蛋白表达。结果与结论:(1)对照组细胞数量较多,呈拥挤状,排列不齐,呈纤维样生长;实验组细胞多呈三角形或多边形,集落状生长,呈铺路石子样;(2)对照组CD31为阴性表达,CD105为阳性表达且阳性率为99.7%,表明细胞仍呈现骨髓间充质干细胞的表型。实验组CD31表达明显上升,阳性率为30.33%;CD105表达下降,阳性率为58.11%,呈现典型的内皮细胞表型;(3)与对照组比较,实验组内皮素、血管内皮细胞生长因子及内皮型一氧化氮合成酶表达增高(P<0.01),前列环素表达降低(P<0.01);(4)结果表明在低氧环境下,血管内皮细胞生长因子具有促使人骨髓间充质干细胞向血管内皮样细胞分化的能力。
BACKGROUND： It has been found that vascular endothelial growth factor can induce the differentiation of bone marrow mesenchymal stem cells into endothelial cells, but can the vascular endothelial growth factor gene promote the differentiation of bone marrow mesenchymal stem cells into vascular endothelial cells in the damaged organ under the hypoxic environment？OBJECTIVE：To observe whether human bone marrow mesenchymal stem cells induced by vascular endothelial growth factor could differentiate into vascular endothelial cells under hypoxia.METHODS：The third passage of human bone marrow mesenchymal stem cells were cultured in vitro.Cells in the control group were cultured with conventional culture medium,while those in experimental group were cultured with adenovirus vector containing vascular endothelial growth factor in 5%O2.After 2 weeks of culture,morphological observation and surface-related molecular detection were performed.The levels of vascular endothelial growth factor and endothelial nitric oxide synthase were detected by ELISA.The expression of endothelin and prostacyclin was detected by RT-PCR and western blot assay.RESULTS AND CONCLUSION：（1）The number of cells in the control group was more than that in the experimental group.The cells in the control group were crowded and arranged irregularly,showing a fiber-like growth,while those in the experimental group were mostly triangular or polygonal,exhibiting a colony-like growth.（2）CD31 was negative in the control group,while CD105 was positive and the positive rate was 99.7%,indicating that the cells still showed the phenotype of bone marrow mesenchymal stem cells.The positive rate of CD31 was significantly increased to 30.33%in the experimental group and the positive rate of CD105 expression was decreased to 58.11%,indicating a typical phenotype of endothelial cells.（3）Compared with the control group,the expression of endothelin,vascular endothelial growth factor and endothelial nitric oxide synthase increased significantly in the experimental group（P〈0.05）,and the expression of prostacyclin decreased significantly（P〈0.05）.All these findings indicate that vascular endothelial growth factor can promote the differentiation of human bone marrow mesenchymal stem cells into vascular endothelial cells under hypoxia.