miR-222-3p 下调通过 ADIPOR1 表达增加诱导的 AMKP 激活促使内皮祖细胞迁移、侵袭和募集。
MiR-222-3p downregulation prompted the migration, invasion and recruitment of endothelial progenitor cells via ADIPOR1 expression increase-induced AMKP activation.
机构信息
Department of Cardiovascular Medicine, The First Affiliated Hospital of Chongqing Medical University, No. 1, Youyi Road, Yuanjiagang, Yuzhong District, Chongqing 400016, China.
Department of Heart and Vascular Surgery, Cardiovascular Disease Center, The Third Affiliated Hospital of Chongqing Medical University, No. 1 Shuanghu Branch Road, Huixing Street, Yubei District, Chongqing 401120, China.
出版信息
Microvasc Res. 2021 May;135:104134. doi: 10.1016/j.mvr.2021.104134. Epub 2021 Jan 9.
BACKGROUND
Clinical data show that aneurysm rupture causes high mortality in aged men. MicroRNAs (miRNAs) were reported to regulate endothelial progenitor cells (EPCs) which play a vital role in repairing endothelial damage and maintaining vascular integrity. This study identified a novel miRNA regulator for the functions of EPCs in aneurysm repair.
METHODS
Abdominal aortic aneurysm (AAA) model was established on Sprague-Dawley rats which later underwent antagomiR-222 treatment. The histopathological changes of AAA rats were examined by hematoxylin-eosin staining. Flow cytometry was performed to quantify EPCs in peripheral blood and identify EPCs isolated from the rat femur. The potential target of miR-222-3p was predicted by TargetScan v7.2 and validated by Dual-luciferase reporter assay. The effects of miR-222-3p and ADIPOR1 on the migration, invasion and tube formation of EPCs were evaluated by wound healing, Transwell and tube formation assays. The expressions of miR-222-3p and ADIPOR1 in aortic aneurysm tissues and EPCs were assessed by qRT-PCR or Western blot.
RESULTS
AAA exhibited histopathological abnormality, a decreased number of EPCs in the peripheral blood and an increased miR-222-3p expression. AntagomiR-222 injection reversed all these phenomena in AAA rats. Upregulating miR-222-3p expression inhibited the migration, invasion, and tube formation of EPCs, and the expressions of ADIPOR1 and phosphorylated-AMKP, while downregulating miR-222-3p expression exerted opposite effects in EPCs. ADIPOR1 was identified as a target gene of miR-222-3p. Overexpressing ADIPOR1 abrogated the effects of miR-222-3p upregulation on EPCs.
CONCLUSION
Downregulated miR-222-3p prompted the migration, invasion and recruitment of EPCs by targeting ADIPOR1-induced AMKP activation.
背景
临床数据表明，动脉瘤破裂会导致老年男性死亡率升高。有研究报道，微小 RNA（miRNA）可调节内皮祖细胞（EPCs），EPCs 在修复内皮损伤和维持血管完整性方面发挥着重要作用。本研究旨在鉴定一种新型 miRNA 调节剂，用于调控动脉瘤修复过程中 EPCs 的功能。
方法
建立 Sprague-Dawley 大鼠腹主动脉瘤（AAA）模型，随后进行 antagomiR-222 治疗。采用苏木精-伊红（H&E）染色观察 AAA 大鼠的组织病理学变化。采用流式细胞术检测外周血中的 EPCs 数量，并从大鼠股骨中分离鉴定 EPCs。通过 TargetScan v7.2 预测 miR-222-3p 的潜在靶标，并通过双荧光素酶报告实验进行验证。采用划痕愈合实验、Transwell 实验和管形成实验评估 miR-222-3p 和 ADIPOR1 对 EPC 迁移、侵袭和管形成的影响。采用 qRT-PCR 或 Western blot 检测主动脉瘤组织和 EPCs 中 miR-222-3p 和 ADIPOR1 的表达。
结果
AAA 表现出组织病理学异常、外周血中 EPC 数量减少和 miR-222-3p 表达增加。在 AAA 大鼠中，antagomiR-222 注射逆转了所有这些现象。上调 miR-222-3p 表达抑制了 EPC 的迁移、侵袭和管形成，以及 ADIPOR1 和磷酸化-AMKP 的表达，而下调 miR-222-3p 表达则在 EPC 中产生相反的效果。ADIPOR1 被鉴定为 miR-222-3p 的靶基因。过表达 ADIPOR1 可消除 miR-222-3p 上调对 EPC 的作用。
结论
下调的 miR-222-3p 通过靶向 ADIPOR1 诱导的 AMKP 激活促进 EPC 的迁移、侵袭和募集。
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