IL-1β promotes hypoxic vascular endothelial cell proliferation through the miR-24-3p/NKAP/NF-κB axis.

Jiangnan Huang,Yueying Liu,Zhiyuan Jiang,Hui Wang,Lang Li,Yumei Li,Lingjun Wu,Siwen Ma

doi:10.1042/bsr20212062

Abstract

Purpose: Our previous data indicated that miR-24-3p is involved in the regulation of vascular endothelial cell (EC) proliferation and migration/invasion. However, whether IL-1β affects hypoxic HUVECs by miR-24-3p is still unclear. Therefore, the present study aimed to investigate the role and underlying mechanism of interleukin 1β (IL-1β) in hypoxic HUVECs.Methods: We assessed the mRNA expression levels of miR-24-3p, hypoxia-inducible factor-1α (HIF1A) and NF-κB-activating protein (NKAP) by quantitative real-time polymerase chain reaction (RT-qPCR). ELISA measured the expression level of IL-1β. Cell counting kit-8 (CCK-8) assays evaluated the effect of miR-24-3p or si-NKAP+miR-24 on cell proliferation (with or without IL-1β). Transwell migration and invasion assays were used to examine the effects of miR-24-3p or si-NKAP+miR-24-3p on cell migration and invasion (with or without IL-1β). Luciferase reporter assays were used to identify the target of miR-24-3p.Results: We demonstrated that in acute myocardial infarction (AMI) patient blood samples, the expression of miR-24-3p is down-regulated, the expression of IL-1β or NKAP is up-regulated, and IL-1β or NKAP is negatively correlated with miR-24-3p. Furthermore, IL-1β promotes hypoxic HUVECs proliferation by down-regulating miR-24-3p. In addition, IL-1β also significantly promotes the migration and invasion of hypoxic HUVECs; overexpression of miR-24-3p can partially rescue hypoxic HUVECs migration and invasion. Furthermore, we discovered that NKAP is a novel target of miR-24-3p in hypoxic HUVECs. Moreover, both the overexpression of miR-24-3p and the suppression of NKAP can inhibit the NF-κB/pro-IL-1β signaling pathway. However, IL-1β mediates suppression of miR-24-3p activity, leading to activation of the NKAP/NF-κB pathway. In conclusion, our results reveal a new function of IL-1β in suppressing miR-24-3p up-regulation of the NKAP/NF-κB pathway.

Highlights

Acute myocardial infarction (AMI) and heart failure (HF) are often the leading causes of death and disability worldwide, especially in developed countries [1,2,3]
Our previous data showed that miR-24 inhibits the proliferation of HUVECs [22], which led us to hypothesize that miR-24-3p could affect hypoxic HUVECs and play an important role in angiogenesis
After transfection with miR-NC or miR-24-3p, the effect of CoCl2 on hypoxia-inducible factor-1α (HIF1A) mRNA in HUVECs was analyzed by quantitative real-time polymerase chain reaction (RT-qPCR), with GAPDH used as an internal control

Summary

Introduction

Acute myocardial infarction (AMI) and heart failure (HF) are often the leading causes of death and disability worldwide, especially in developed countries [1,2,3]. AMI is an event of myocardial necrosis that may lead to functional loss in the diastolic and systolic regions and cause arrhythmias in patients [4]. AMI and ischemia–reperfusion injury dysfunction in the myocardium induce an acute inflammatory response, which activates the inflammasome and maturation of proinflammatory cytokines (e.g., interleukin 1β (IL-1β)) [5]. IL-1β induces a further loss of viable myocardium, promoting cardiac dilation and dysfunction in the subacute phase of AMI and suppressing cardiac contractility and β-adrenergic receptor responsiveness [7]. Inhibitors of the inflammasome or IL-1β administered during AMI have the potential to prevent adverse cardiac remodeling and HF

Methods

Results

Conclusion