作　　者： (葛权虎）; (吴向未）; (程文哲）; (万龙飞）; (王小义）; (赵伯文）;

机构地区： 石河子大学医学院第一附属医院,新疆石河子832008

出　　处： 《山东医药》 2017年第28期12-15,共4页

摘　　要： 目的探讨小鼠骨髓间充质干细胞(MSCs)对血管内皮祖细胞(EPCs)向内皮细胞(ECs)分化的影响。方法分离、培养及扩增小鼠骨髓MSCs和EPCs。取5×10~5个EPCs接种于含5%FBS的EBM-2培养液,置于Transwel下室(对照组);取5×10~5个EPCs接种于含5%FBS的EBM-2+20 ng/m L血管内皮细胞生长因子(VEGF)的培养液,置于Transwell下室(VEGF组);取5×10~5个MSCs接种于Transwell insert膜上、5×10~5个EPCs接种于Transwell下室(共培养组),培养液同对照组。培养48 h,收集各组下室EPCs,采用RT-PCR法检测内皮型一氧化氮合成酶(eNOS)和激酶插入结构域受体(KDR)mRNA表达;ELISA法检测各组细胞培养液上清VEGF含量。提取MSC条件培养基(MSCCM)及LG-DMEM基础培养基,ELISA法检测VEGF含量。取传3代EPCs,分别用LGDMEM、MSCCM、MSCCM+IgG、MSCCM+VEGF抗体的培养液培养48 h(分别设为LG-DMEM组、MSCCM组、MSCCM+IgG组、MSCCM+VEGF抗体组),收集细胞,采用RT-PCR法检测各组eNOS、KDR mRNA表达。结果共培养组eNOS、KDR mRNA相对表达量明显高于对照组和VEGF组(P<0.05或<0.01);共培养组、VEGF组培养液上清VEGF含量均明显高于对照组(P均<0.01)。MSCCM中VEGF含量较LG-DMEM基础培养基明显增加(P<0.01)。MSCCM组和MSCCM+IgG组eNOS、KDR mRNA相对表达量较LG-DMEM组、MSCCM+VEGF抗体组均明显增加(P均<0.01)。结论小鼠骨髓MSCs可能通过旁分泌VEGF促进血管EPCs向ECs分化。 Objective To investigate the effect of mesenchymal stem cells（ MSCs） on the differentiation of endothelial progenitor cells（ EPCs） into endothelial cells（ ECs）. Methods MSCs and EPCs were isolated,cultured,and amplified from murine bone marrow in vitro. MSCs and EPCs were cultured in Transwell co-culture system. The experiment was totally divided into three groups,EPCs cultured group（ control group）,co-culture group,and vascular endothelial growth factor group（ VEGF group）. After being cultured for 48 h,EPCs were collected and the expression of the specific markers of eNOS and KDR was measured by RT-PCR; the expression of VEGF in the culture medium of each group was detected by ELISA.MSC conditioned medium（ MSCCM） was collected. EPCs which were at passage 3 were cultured in the medium of LGDMEM,MSCCM,MSCCM ＋ IgG,MSCCM ＋ VEGF neutralizing antibody（ Anti-VEGF）,respectively,which were taken as the following four groups,LG-DMEM group,MSCCM group,MSCCM ＋ IgG group,and MSCCM ＋ Anti-VEGF group. After being cultured for 48 h,EPCs were collected and the expression of eNOS and KDR was measured by RT-PCR. Results The mRNA expression of the specific markers was significantly higher in the co-culture group and VEGF group than that in the control group（ P〈0. 05 or 0. 01）. The expression of VEGF in the co-culture group and VEGF group was significantly increased as compared with that of the control group（ P〈0. 01）. The mRNA expression of the specific markers in the MSCCM group and MSCCM ＋ IgG group was significantly higher than that of the LG-DMEM group and MSCCM ＋ Anti-VEGF group（ P〈0. 01）. Conclusion MSCs can promote the differentiation of EPCs into ECs through paracrine factors such as VEGF.

关 键 词： 骨髓间充质干细胞 血管内皮祖细胞 内皮细胞 细胞分化 血管内皮细胞生长因子