miR-222-3p 的上调通过靶向 STAT3 缓解主动脉夹层症状。

Upregulation of miR-222-3p alleviates the symptom of aortic dissection through targeting STAT3.

机构信息

Department of Cardiovascular Surgery, Fujian Medical University Union Hospital, Fuzhou 350001, China; Key Laboratory of Cardio-Thoracic Surgery (Fujian Medical University), Fujian Province University, Fuzhou, Fujian 350001, China; Fujian Provincial Special Reserve Talent Laboratory, Fuzhou, Fujian 350108, China.

出版信息

Life Sci. 2022 Dec 1;310:121051. doi: 10.1016/j.lfs.2022.121051. Epub 2022 Oct 8.

DOI:10.1016/j.lfs.2022.121051

PMID:36216078

Abstract

OBJECTIVE

This study sought to investigate the differentially expressed miRNAs in Aortic dissection (AD) and explore the downstream mechanisms in regulating AD.

METHODS

Exosomes of AD patients and healthy people were isolated by differential centrifugation, and the differentially expressed miRNAs were evaluated by RNA sequencing. The downstream target of miR-222-3p was predicted by bioinformatics method and validated by dual-luciferase assay. Angiotensin II and Promethazine were used to establish AD mouse model and platelet-derived growth factor BB (PDGF-BB) was used to induce human vascular smooth muscle cells (HVSMCs) to elucidate the effect of miR-222-3p upregulation on AD in vivo and in vitro. The relative level of miR-222-3p was evaluated by RT-qPCR. The level of several proteins was investigated by Western blot. Immunofluorescence staining was used to detect the stress fiber formation. Cell migration was evaluated by wound healing and Transwell assay. The proliferation, cell cycle and apoptosis of HVSMCs were assessed by CCK-8 and flow cytometry, respectively.

RESULTS

MiR-222-3p was downregulated in AD and PDGF-BB induced HVSMCs. The upregulation of miR-222-3p alleviated the symptom of AD in vivo by targeting STAT3, and inhibited stress fiber formation, abnormal migration, proliferation and apoptosis of HVSMCs induced by PDGF-BB by regulating the expression of α-SMA, SM22α, MMP2, MMP9 and p-Smad2.

CONCLUSION

The upregulation of miR-222-3p attenuates the progression of AD. Our study provides a theoretical basis for exploring new strategies against AD.

摘要

目的

本研究旨在探讨主动脉夹层（AD）中差异表达的 miRNAs，并探索其调节 AD 的下游机制。

方法

通过差速离心法分离 AD 患者和健康人的外泌体，通过 RNA 测序评估差异表达的 miRNAs。通过生物信息学方法预测 miR-222-3p 的下游靶标，并通过双荧光素酶报告基因实验进行验证。使用血管紧张素 II 和奋乃静建立 AD 小鼠模型，使用血小板衍生生长因子 BB（PDGF-BB）诱导人血管平滑肌细胞（HVSMCs），以阐明 miR-222-3p 上调对 AD 体内和体外的影响。通过 RT-qPCR 评估 miR-222-3p 的相对水平。通过 Western blot 检测几种蛋白质的水平。免疫荧光染色检测应激纤维形成。通过划痕愈合和 Transwell 测定评估细胞迁移。通过 CCK-8 和流式细胞术分别评估 HVSMCs 的增殖、细胞周期和凋亡。

结果

miR-222-3p 在 AD 中下调，在 PDGF-BB 诱导的 HVSMCs 中下调。miR-222-3p 的上调通过靶向 STAT3 缓解 AD 症状，通过调节α-SMA、SM22α、MMP2、MMP9 和 p-Smad2 的表达，抑制 PDGF-BB 诱导的 HVSMCs 应激纤维形成、异常迁移、增殖和凋亡。