七氟醚预处理通过调节 NORAD/miR-144-3p 轴减轻缺氧/复氧诱导的心肌细胞损伤。

Sevoflurane preconditioning attenuates myocardial cell damage caused by hypoxia and reoxygenation via regulating the NORAD/miR-144-3p axis.

机构信息

Department of Anesthesiology, Affiliated Hospital of North Sichuan Medical College, Nanchong, China.

Cardiology Department, The Second Affiliated Hospital of Hunan University of Chinese Medicine, Changsha, China.

出版信息

Hum Exp Toxicol. 2024 Jan-Dec;43:9603271241297883. doi: 10.1177/09603271241297883.

DOI:10.1177/09603271241297883

PMID:39586668

Abstract

OBJECTIVE

This study aimed to investigate the function and mechanism of lncRNA NORAD in Sevoflurane (Sev) protection against myocardial hypoxia-reoxygenation (H/R).

METHODS

Preprocess rat cardiomyocytes H9c2 cells with Sev at concentrations of 0.5%, 1.0%, and 1.5%, and subjected them to H/R treatment. qRT-PCR was used to detect levels of NORAD and miR-144-3p. Measure concentrations of the inflammatory cytokines IL-6, TNF-α, and IL-10, as well as cardiac injury markers cTnI, CK-MB, and LDH using ELISA. Assess cell proliferation and apoptosis using CCK-8 and flow cytometry. Perform dual-luciferase reporter assay and RIP assay to validate the targeting relationship between NORAD and miR-144-3p.

RESULTS

H/R induced inhibition of cell proliferation, increase in apoptosis, and production of IL-6, TNF-α, CK-MB, LDH, and cTnI were significantly attenuated by Sev. As hypoxic treatment time lengthened, the NORAD levels in myocardial cells showed an increase, with Sev pretreatment being able to suppress the NORAD levels elevation. The overexpression of NORAD notably weakened the cardioprotective effect of Sev. NORAD targetedly binds to miR-144-3p and negatively regulates miR-144-3p. Increased miR-144-3p levels inhibited the antagonistic effect of NORAD on the cardioprotective effects of Sev.

CONCLUSION

The current study confirmed that sevoflurane attenuated H/R-induced cardiomyocyte injury via the NORAD/miR-144-3p axis.

摘要

目的

本研究旨在探讨长链非编码 RNA（lncRNA）NORAD 在七氟醚（Sev）对心肌缺氧复氧（H/R）保护中的作用和机制。

方法

用不同浓度（0.5%、1.0%和 1.5%）的 Sev 预处理大鼠心肌细胞 H9c2，然后进行 H/R 处理。qRT-PCR 检测 NORAD 和 miR-144-3p 的水平。采用 ELISA 法检测炎症细胞因子 IL-6、TNF-α和 IL-10 的浓度，以及心肌损伤标志物 cTnI、CK-MB 和 LDH。用 CCK-8 和流式细胞术检测细胞增殖和凋亡。通过双荧光素酶报告基因和 RIP 实验验证 NORAD 与 miR-144-3p 的靶向关系。

结果

H/R 诱导的细胞增殖抑制、凋亡增加以及 IL-6、TNF-α、CK-MB、LDH 和 cTnI 的产生均被 Sev 显著减弱。随着缺氧处理时间的延长，心肌细胞中的 NORAD 水平升高，Sev 预处理能够抑制 NORAD 水平的升高。NORAD 的过表达显著减弱了 Sev 的心脏保护作用。NORAD 靶向结合 miR-144-3p，并负调控 miR-144-3p。miR-144-3p 水平的升高抑制了 NORAD 对 Sev 心脏保护作用的拮抗作用。