长链非编码 RNA NEAT1 通过海绵吸附 miR-144-3p 促进脓毒症诱导的心肌细胞损伤的进展。

LncRNA NEAT1 promotes the progression of sepsis-induced myocardial cell injury by sponging miR-144-3p.

机构信息

Department of Hematology, The First Affiliated Hospital of Soochow University, Suzhou, Jiangsu, China.

出版信息

Eur Rev Med Pharmacol Sci. 2020 Jan;24(2):851-861. doi: 10.26355/eurrev_202001_20069.

DOI:10.26355/eurrev_202001_20069

PMID:32016991

Abstract

OBJECTIVE

Sepsis is a systemic inflammatory response that can lead to the dysfunction of many organs, including the cardiac one. Long noncoding RNAs (lncRNAs) have been shown to be involved in multiple organ injuries induced by sepsis. However, the regulatory effect of nuclear enriched abundant transcript 1 (NEAT1) on sepsis-induced myocardial injury remains to be explored.

MATERIALS AND METHODS

The sepsis models of myocardial cell injury were constructed using lipopolysaccharide (LPS). Cell counting kit-8 (CCK-8) assay was used to detect cell viability. Flow cytometry was performed to assess cell apoptosis. Moreover, the levels of apoptosis-related and nuclear factor-kappa B (NF-κB) signaling pathway-related proteins were evaluated by Western blot (WB) analysis. Besides, the contents of inflammatory cytokines were tested by enzyme-linked immunosorbent assay (ELISA). The expression levels of NEAT1 and microRNA-144-3p (miR-144-3p) were determined by quantitative Real Time-Polymerase Chain Reaction (qRT-PCR). In addition, Dual-Luciferase reporter and RNA immunoprecipitation (RIP) assays were used to verify the interaction between NEAT1 and miR-144-3p.

RESULTS

LPS could induce myocardial cell injury to construct sepsis models. NEAT1 was upregulated in LPS-treated myocardial cells, and its knockdown promoted viability, suppressed apoptosis, and relieved inflammatory response in LPS-induced myocardial cell injury. MiR-144-3p was downregulated in LPS-treated myocardial cells, and the effect of its overexpression on LPS-induced myocardial cell injury was similar to the effect of NEAT1 knockdown. Besides, miR-144-3p could be sponged by NEAT1, and its inhibitor could reverse the effect of NEAT1 knockdown on LPS-induced myocardial cell injury. Moreover, NEAT1 and miR-144-3p could regulate the activity of NF-κB signaling pathway.

CONCLUSIONS

LncRNA NEAT1 could interact with miR-144-3p to regulate sepsis-induced myocardial cell injury through the NF-κB signaling pathway, which might provide a new theoretical basis for the study on the effect of sepsis treatment.

摘要

目的

脓毒症是一种全身性炎症反应，可导致包括心脏在内的多个器官功能障碍。长链非编码 RNA（lncRNA）已被证明参与脓毒症引起的多种器官损伤。然而，核富集丰富转录本 1（NEAT1）对脓毒症诱导的心肌损伤的调节作用仍有待探索。

材料和方法

使用脂多糖（LPS）构建心肌细胞损伤的脓毒症模型。使用细胞计数试剂盒-8（CCK-8）测定法检测细胞活力。通过流式细胞术评估细胞凋亡。此外，通过 Western blot（WB）分析评估凋亡相关和核因子-κB（NF-κB）信号通路相关蛋白的水平。此外，通过酶联免疫吸附测定（ELISA）检测炎症细胞因子的含量。通过定量实时聚合酶链反应（qRT-PCR）测定 NEAT1 和 microRNA-144-3p（miR-144-3p）的表达水平。此外，使用双荧光素酶报告和 RNA 免疫沉淀（RIP）测定验证 NEAT1 和 miR-144-3p 之间的相互作用。

结果

LPS 可诱导心肌细胞损伤构建脓毒症模型。LPS 处理的心肌细胞中 NEAT1 上调，其敲低可促进活力，抑制凋亡，并缓解 LPS 诱导的心肌细胞损伤中的炎症反应。miR-144-3p 在 LPS 处理的心肌细胞中下调，其过表达对 LPS 诱导的心肌细胞损伤的作用与 NEAT1 敲低的作用相似。此外，miR-144-3p 可以被 NEAT1 海绵化，其抑制剂可以逆转 NEAT1 敲低对 LPS 诱导的心肌细胞损伤的作用。此外，NEAT1 和 miR-144-3p 可以调节 NF-κB 信号通路的活性。