血管细胞外基质胶促进骨髓CD34+祖细胞分化为内皮细胞的实验研究

doi:10.11958/20221138

天津医药 ›› 2023, Vol. 51 ›› Issue (3): 225-229.doi: 10.11958/20221138

• 细胞与分子生物学 • 下一篇

血管细胞外基质胶促进骨髓CD34⁺祖细胞分化为内皮细胞的实验研究

左芯萌¹(), 王振华², 高利平^△()

1 徐州医科大学基础医学院生理学教研室（邮编221004）
2 上海交通大学医学院附属仁济医院心外科

收稿日期:2022-07-19 修回日期:2022-09-28 出版日期:2023-03-15 发布日期:2023-03-02
通讯作者: 高利平 E-mail:2444392396@qq.com;pandag2011@sina.com
作者简介:左芯萌（1997），女，硕士在读，主要从事心血管损伤及修复方面研究。E-mail：2444392396@qq.com
基金资助:
徐州市科技计划项目(KC20155)

Experimental study of vascular extracellular matrix gel promoting the differentiation of bone marrow CD34⁺ progenitor cells into endothelial cells

ZUO Xinmeng¹(), WANG Zhenhua², GAO Liping^△()

1 Department of Physiology, School of Basic Medicine, Xuzhou Medical University, Xuzhou 221004, China
2 Department of Cardiovascular Surgery, Ren Ji Hospital, School of Medicine, Shanghai Jiao Tong University

Received:2022-07-19 Revised:2022-09-28 Published:2023-03-15 Online:2023-03-02
Contact: GAO Liping E-mail:2444392396@qq.com;pandag2011@sina.com

左芯萌, 王振华, 高利平. 血管细胞外基质胶促进骨髓CD34⁺祖细胞分化为内皮细胞的实验研究[J]. 天津医药, 2023, 51(3): 225-229.

ZUO Xinmeng, WANG Zhenhua, GAO Liping. Experimental study of vascular extracellular matrix gel promoting the differentiation of bone marrow CD34⁺ progenitor cells into endothelial cells[J]. Tianjin Medical Journal, 2023, 51(3): 225-229.

摘要/Abstract

摘要： 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, CD144, vascular endothelial growth factor receptor 2 (VEGFR2), CD133 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, CD144 and VEGFR2 expressing cells and the number of phagocytosing Dil-acLDL cells were also increased in the ECM group, while the positive rates of CD133 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 endothelial cells, which may provide a new strategy for in situ endothelialization of cardiovascular implants.

关键词: 细胞外基质, 内皮细胞, 细胞分化, CD34⁺祖细胞, 内皮化

Key words: extracellular matrix, endothelial cells, cell differentiation, CD34⁺ progenitor cells, endothelialization

中图分类号:

R331.3

图/表 9

图1 患者主动脉组织（标尺=1 cm）

Fig.1 A patient’s aortic tissue (scale bar=1 cm)

图2 人主动脉组织HE及DAPI染色（×40）

Fig.2 HE and DAPI staining of human aortic tissue (×40)

图3 CD34+祖细胞纯度

Fig.3 Purity of CD34+ progenitor cells

图4 CD34+祖细胞在FN和ECM胶上的生长（×20）

Fig.4 Growth of CD34+ progenitor cells on FN and ECM gel (×20)

图5 CD34+祖细胞CD31、eNOS及VWF的mRNA表达

Fig.5 mRNA expression of CD31, eNOS and VWF in CD34+ progenitor cells in two groups

表1 2组CD31、eNOS、VWF mRNA表达水平比较 (n=3,$\bar{x}$±s)

Tab.1 Comparison of mRNA expression levels of CD31, eNOS and VWF between the two groups

组别	CD31	eNOS	VWF
FN组	0.858±0.044	0.462±0.137	0.634±0.135
ECM组	0.992±0.077	0.630±0.191	0.745±0.148
t	4.522^＊	5.363^＊	14.520^＊＊

图6 2组分化相关标志物阳性的CD34+祖细胞百分率

Fig.6 The percentage of CD34+ progenitor cells with positive differentiation-related markers in two groups

图7 CD34+祖细胞吞噬Dil-acLDL（×40）

Fig.7 Phagocytosis for DiI-ACLDL by CD34+ progenitor cells (×40)

表2 2组分泌促血管生成因子水平比较 (n=3,$\bar{x}$±s)

Tab.2 Comparison of levels of secreted angiogenic factors between the two groups

组别	Angiogenin	FGF-7	GM-CSF	Leptin	PDGF-BB
FN组	184.7±24.2	425.7±34.3	104.3±22.0	548.7±74.2	634.7±85.1
ECM组	473.3±46.1	767.3±45.0	192.0±29.5	848.7±136.5	1 009.2±199.1
t	10.810^＊＊	12.090^＊＊	16.730^＊	8.294^＊	5.662^＊

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血管细胞外基质胶促进骨髓CD34⁺祖细胞分化为内皮细胞的实验研究

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血管细胞外基质胶促进骨髓CD34+祖细胞分化为内皮细胞的实验研究

Experimental study of vascular extracellular matrix gel promoting the differentiation of bone marrow CD34+ progenitor cells into endothelial cells

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血管细胞外基质胶促进骨髓CD34⁺祖细胞分化为内皮细胞的实验研究

Experimental study of vascular extracellular matrix gel promoting the differentiation of bone marrow CD34⁺ progenitor cells into endothelial cells