摘要
目的研究血管细胞外基质(ECM)胶促进骨髓CD34^(+)祖细胞分化为内皮细胞的作用。方法将人主动脉组织脱细胞后得到ECM,胃蛋白酶消化后得到ECM胶。HE染色和DAPI染色观察主动脉组织脱细胞效果。将小鼠骨髓CD34^(+)祖细胞分为2组:纤连蛋白(FN)组置于涂有FN的培养板中培养;ECM组置于板底铺有人主动脉ECM胶的培养板中培养。逆转录聚合酶链反应(RT-PCR)检测CD34^(+)祖细胞CD31、内皮型一氧化氮合酶(e NOS)及血管性血友病因子(VWF)mRNA表达;流式细胞术检测CD34^(+)祖细胞表面标志物CD31、CD144、血管内皮生长因子受体2(VEGFR2)、CD133及CD45阳性细胞百分率;荧光显微镜下观察吞噬荧光素标记的乙酰化低密度脂蛋白(DiIacLDL)的细胞数;蛋白芯片检测细胞分泌的促血管生成因子含量。结果DAPI和HE染色均显示人主动脉组织脱细胞前含有大量细胞核,而脱细胞后得到的血管ECM中不含细胞核。与FN组相比,ECM组CD34^(+)祖细胞CD31、eNOS及VWF mRNA表达升高,表达CD31、CD144及VEGFR2的阳性细胞率及吞噬Dil-acLDL的细胞数量增加,而表达CD133和CD45的阳性细胞率减少(P<0.05)。ECM组分泌血管生成素、成纤维细胞生长因子7、粒细胞-巨噬细胞集落刺激因子、瘦素及血小板源性生长因子BB较FN组增多(P<0.05)。结论血管ECM胶可促进骨髓CD34^(+)祖细胞分化为内皮细胞,可能为心血管植入物原位内皮化提供新策略。
Objective To investigate the role of vascular extracellular matrix(ECM)gel in promoting the differentiation of bone marrow CD34^(+)progenitor cells into endothelial cells.Methods ECM was obtained from the decellularized aortic tissue,and vascular ECM gel was prepared by enzyme digestion of vascular ECM.HE staining and DAPI staining were used to observe the decellularization effect of aortic tissue.Mouse bone marrow CD34^(+)progenitor cells were divided into two groups:the fibronectin(FN)group was cultured in plates coated with FN,and the ECM group was cultured in plates coated with human aorta ECM gel at the bottom of the plate.The mRNA expression of CD31,endothelial nitric oxide synthase(eNOS)and Von willebrand factor(VWF)were detected by RT-PCR in both groups.Flow cytometry was used to detect the percentage rates of positive CD31,CD 144,vascular endothelial growth factor receptor 2(VEGFR2),CD 133 and CD45 positive cells.The number of phagocytic fluorescein labeled acetylated low-density lipoprotein(DiI-acLDL)cells was observed under fluorescence microscope.The content of proangiogenic factors secreted by cells was detected by protein microarray.Results Both HE and DAPI staining showed that human aortic tissue contained a large number of nuclei before decellularization,while the vascular ECM obtained after decellularization did not contain nuclei.Compared with the FN group,the expression levels of CD31,eNOS and VWF mRNA in CD34^(+)progenitor cells were increased,the positive rates of CD31,CD 144 and VEGFR2 expressing cells and the number of phagocytosing Dil-acLDL cells were also increased in the ECM group,while the positive rates of CD 133 and CD45 expressing cells decreased(P<0.05).The secretion of angiopoietin,fibroblast growth factor 7,granulocyte macrophage colony stimulating factor,leptin and platelet-derived growth factor BB were significantly higher in the ECM group than those of the FN group(P<0.05).Conclusion Vascular ECM gel promotes the differentiation of bone marrow CD34^(+)progenitor cells into end
作者
左芯萌
王振华
高利平
ZUO Xinmeng;WANG Zhenhua;GAO Liping(Department of Physiology,School of Basic Medicine,Xuzhou Medical University,Xuzhou 221004,China;Department of Cardiovascular Surgery,Ren Ji Hospital,School of Medicine,Shanghai Jiao Tong University)
出处
《天津医药》
CAS
北大核心
2023年第3期225-229,共5页
Tianjin Medical Journal
基金
徐州市科技计划项目(KC20155)。
关键词
细胞外基质
内皮细胞
细胞分化
CD34+祖细胞
内皮化
extracellular matrix
endothelial cells
cell differentiation
CD34+progenitor cells
endothelialization
