广东畜牧兽医科技2024,Vol.49Issue(3) :25-32.DOI:10.19978/j.cnki.xmsy.2024.03.04

cfa-miR-21对犬血管内皮细胞增殖、迁移的调控作用及机制

Regulation and Mechanism of cfa-miR-21 Carried by Canine Umbilical Cord Mesenchymal Stem Cell Exosomes on Proliferation and Migration of Canine Vascular Endothelial Cells

张妙齐 赖健仪 詹小舒 王丙云 李东升 阮慧敏
广东畜牧兽医科技2024,Vol.49Issue(3) :25-32.DOI:10.19978/j.cnki.xmsy.2024.03.04

cfa-miR-21对犬血管内皮细胞增殖、迁移的调控作用及机制

Regulation and Mechanism of cfa-miR-21 Carried by Canine Umbilical Cord Mesenchymal Stem Cell Exosomes on Proliferation and Migration of Canine Vascular Endothelial Cells

张妙齐 1赖健仪 1詹小舒 1王丙云 1李东升 2阮慧敏2
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作者信息

  • 1. 佛山科学技术学院生命科学与工程学院,广东佛山 528200
  • 2. 维赛(佛山)生物科技有限公司,广东佛山 528200
  • 折叠

摘要

为研究犬脐带间充质干细胞外泌体(UC-MSC-Exo)中携带的cfa-miR-21促进犬血管内皮细胞(VECs)增殖及迁移的作用机制,该实验以犬VECs为模型,首先通过qRT-PCR筛选出cfa-miR-21最佳转染方案,然后使用EdU法和划痕实验,检测cfa-miR-21对细胞的增殖及迁移的调控作用;通过生物信息学分析软件预测靶基因,并利用双荧光素酶报告基因系统和qRT-PCR进行验证,再通过Western Blot检测下游ERK信号通路蛋白的表达情况.实验结果表明,cfa-miR-21可促进犬VECs的增殖和迁移,其可通过靶向作用于SPRY1,从而激活ERK信号通路,发挥其在血管生成中的生物功能.该研究为miRNAs在血管生成中的作用提供了见解,揭示了间充质干细胞(MSCs)的功能机制,并可能有助于未来的药物开发.

Abstract

In order to study the mechanism of cfa-miR-21 carried by canine umbilical cord mesenchymal stem cell exosomes(UC-MSC-Exo)in promoting the proliferation and migration of canine vascular endothelial cells(VECs),the following studies were carried out in this experiment.In this study,canine VECs was used as a model.The best transfection scheme of cfa-miR-21 transfection reagent was screened by qRT-PCR,and the regulatory effect of cfa-miR-21 on cell proliferation and migration was detected by EdU cell proliferation imaging and scratch test.The target gene was predicted by bioinformatics analysis software,and the target gene was verified by dual luciferase reporter gene system and qRT-PCR.Then the expression of downstream ERK signal pathway protein was detected by Western Blot.The results suggest that cfa-miR-21 can promote the proliferation and migration of canine vascular endothelial cells,and cfa-miR-21 can exert its biological function by targeting SPRY1 and activating ERK signal pathway.This study offers new insights into the role of miRNAs in regulating angiogenesis,uncovering the functional mechanism of MSCs and potentially contributing to future drug development.

关键词

/血管内皮细胞/脐带间充质干细胞外泌体/miR-21/细胞增殖/细胞迁移

Key words

Canine/Vascular endothelial cells/Umbilical cord mesenchymal stem cell exosomes/miR-21/Cells proliferation/Cells migration

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基金项目

广东省普通高等学校动物干细胞工程技术研究中心项目(2021GCZX006)

出版年

2024
广东畜牧兽医科技
广东省畜牧兽医学会 广东省农业科学院畜牧研究所 广东省农业科学院兽医研究所

广东畜牧兽医科技

影响因子:0.29
ISSN:1005-8567
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