miR-148a-3p 调控血管生成的体内外表型鉴定

摘要/Abstract

摘要：

目的　明确miR-148a-3p调控血管生成的表型。方法　提取miR-148a基因敲除小鼠的原代组织，结合免疫组织化学染色，探究miR-148a敲除对血管生成的影响；通过miRNA…mimic转染，在人脐静脉血管内皮细胞（hUVECs）中过表达miR-148a-3p，结合Matrigel基质胶管腔形成实验、细胞迁移实验、荧光定量PCR实验等，从敲除和过表达两方面明晰miR-148a-3p对血管生成的作用。结果　与同窝对照相比，miR-148a基因敲除小鼠的骨骼组织存在H型血管分布异常。离体实验得到了相似的趋势，miR-148a敲除胎鼠的跖骨具有明显更强的血管形成能力。通过miR-148a-3p…mimic过表达血管内皮细胞中的miR-148a-3p表达水平，发现miR-148a-3p过表达不影响hUVECs的增殖能力，但可通过抑制其迁移能力干扰血管生成，影响伪足形成、管腔形成等过程。结论　miR-148a-3p可抑制体内外血管生成，包括细胞出芽、迁移、管腔形成等生物学过程。

关键词: miR-148a-3p, 　血管生成, 　基因敲除小鼠, 　表型鉴定, 　H型血管

Abstract:

Objective　To identify the phenotype of miR-148a-3p in regulating angiogenesis. Methods　The primary

tissues of miR-148a knock-out mice were extracted and immunohistochemical staining was used to investigate the effect of

miR-148a knockout on angiogenesis. By means of miRNA Mimic transfection, miR-148a-3p was overexpressed in human

umbilical vein endothelial cells (hUVECs). Thereafter, the influence of miR-148a-3p overexpression on angiogenesis was

studied in aspects of Matrigel-based tube forming assay, cell proliferation, cell migration, qPCR, etc. The effects of miR-148a-

3p on angiogenesis were clarified in terms of knock-out and overexpression. Results　Compared with heterozygous littermates,

the miR-148a knock-out mice had an abnormal distribution of type-H vessels in the bone tissue. The ex-vivo fetal metatarsal

experiments further confirmed the enhanced angiogenesis ability in absence of miR-148a. The expression level of miR-148a-

3p was overexpressed by miR-148a-3p mimic in vascular endothelial cells. It was found that the overexpression of miR-148a-

3p did not affect the proliferation ability of hUVECs, but could interfere with angiogenesis by inhibiting its migration ability, and

affect the formation of lamellipodia and tubes. Conclusion　miR-148a-3p can inhibit angiogenesis both in-vivo and in-vitro,

including sprouting, migration, and tube formation.

Key words:

miR-148a-3p, 　Angiogenesis, 　Gene knock-out mice, 　Phenotype identification, 　Type H vessels

黄瑾, 韩文卿, 李鑫, 等.

miR-148a-3p 调控血管生成的体内外表型鉴定

[J]. 组织工程与重建外科杂志, 2023, 19(6): 525-.

HUANG Jin, HAN Wenqing, LI Xin, et al.

In vivo and vitro phenotype identification of miR-148a-3p regulating angiogenesis

[J]. Journal of Tissue Engineering and Reconstructive Surgery, 2023, 19(6): 525-.

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