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。
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.
作者
黄瑾
韩文卿
李鑫
陈骁俊
HUANG Jin;HAN Wenqing;LI Xin;CHEN Xiaojun(Department of Central Lab,Shanghai Ninth People’s Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China;Department of Plastic and Reconstructive Surgery,Shanghai Ninth People’s Hospital,Shanghai Jiao Tong University School of Medicine,Shanghai 200011,China)
出处
《组织工程与重建外科》
CAS
2023年第6期525-529,540,共6页
Journal of Tissue Engineering and Reconstructive Surgery
基金
上海市科学技术委员会启明星培育(扬帆专项,23YF1421700)
上海市科学技术委员会临床医学研究中心项目(22MC1940300)。
