MiR-539-5p inhibits the inflammatory injury in septic H9c2 cells by regulating IRAK3

Xiaochen Hu,Hongjun Miao

doi:10.1007/s11033-021-06849-1

Abstract

BackgroundMicroRNAs (miRNAs) have been confirmed to play a potential role in sepsis, but little is known about their role in sepsis-induced cardiomyopathy (SIC).MethodsThe model of septic cardiomyopathy was constructed with H9c2 cells induced by lipopolysaccharide (LPS), and the expression of miR-539-5p was detected by qRT-PCR assay. ELISA, CCK-8, EdU TUNEL analysis were performed to evaluate the role of miR-539-5p in inflammation response, viability, proliferation and apoptosis of LPS-treated H9c2 cells. Moreover, miRWalk and TargetScan prediction, and dual-luciferase reporter gene assays were carried out to predict and confirm the target of miR-539-5p. Furthermore, the effects of target on inflammation response, proliferation and apoptosis of LPS-induced H9c2 cells mediated by miR-539-5p was further explored.ResultsThe expression of miR-539-5p was obviously down-regulated in LPS-induced H9c2 cells. In addition, over-expression of miR-539-5p significantly inhibited the inflammation response, promoted viability and proliferation, and suppressed apoptosis of LPS-treated H9c2 cells. Moreover, interleukin-1 receptor-associated kinase 3 (IRAK3) was verified as a target of miR-539-5p by dual-luciferase reporter gene assay. Besides, IRAK3 was highly expressed in H9c2 cells transfected with miR-539-5p inhibitor detected with qRT-PCR and western blot assays. Furthermore, over-expression of IRAK3 partially weakened the effects of miR-539-5p mimic on the inflammation response, proliferation and apoptosis of LPS-induced H9c2 cells.ConclusionsMiR-539-5p potentially plays an important role in the pathogenesis of LPS-induced sepsis by targeting IRAK3, suggesting that miR-539-5p may be a potential new target for the treatment of LPS-induced sepsis.

Highlights

Sepsis is defined as systemic inflammatory response syndrome (SIRS) caused by infection, and its essence is host autoimmune damage mediated by inflammatory mediators and cytokines [1, 2]
To investigate the possible role of miR-539-5p in sepsisinduced cardiomyopathy, firstly, H9c2 cells were treated with different concentrations of LPS (0.25, 5 and 10 μg/ mL), and the expression of miR-539-5p was evaluated by qRT-PCR assay
Myocardial inhibition occurs in the early stage of sepsis, and it will lead to cardiac insufficiency with the progress of the disease, which is the main cause of death for patients with non-cardiogenic heart disease in intensive care unit [21]

Summary

Introduction

Sepsis is defined as systemic inflammatory response syndrome (SIRS) caused by infection, and its essence is host autoimmune damage mediated by inflammatory mediators and cytokines [1, 2]. Bacterial infection is the most common cause of sepsis [7] Bacterial products, such as endotoxin, exotoxin and bacterial outer wall component-- lipopolysaccharide (LPS), can start the inflammatory process through directly or indirectly stimulating monocytes, polymorph nuclear neutrophils, endothelial cells and other target cells, leading to a series of cascade reactions, and further resulting in multiple organ failure [8, 9]. The effects of target on inflammation response, proliferation and apoptosis of LPS-induced H9c2 cells mediated by miR-539-5p was further explored. Over-expression of miR-539-5p significantly inhibited the inflammation response, promoted viability and proliferation, and suppressed apoptosis of LPS-treated H9c2 cells. Over-expression of IRAK3 partially weakened the effects of miR-539-5p mimic on the inflammation response, proliferation and apoptosis of LPS-induced H9c2 cells. Conclusions MiR-539-5p potentially plays an important role in the pathogenesis of LPS-induced sepsis by targeting IRAK3, suggesting that miR-539-5p may be a potential new target for the treatment of LPS-induced sepsis

Methods

Results

Conclusion