LncRNA-5657 silencing alleviates sepsis-induced lung injury by suppressing the expression of spinster homology protein 2

Int Immunopharmacol. 2020 Nov:88:106875. doi: 10.1016/j.intimp.2020.106875. Epub 2020 Aug 28.

Abstract

Background: LncRNAs are closely associated with many human major diseases, however, the roles of lncRNAs in sepsis-induced lung injury remain unclear.

Methods: High-throughput sequencing was used to detect the changes in lncRNA expression profile in alveolar macrophages after LPS stimulation. The BALF of patients with sepsis-induced lung injury was collected. Rats with CLP-induced septic lung injury were treated with sh-lncRNA-5657 via intravenous injection. NR8383 cells were transfected with lentiviral vectors expressing lncRNA-5657, lncRNA-5657 smart silencer, or si-spns2. The BALF cells expression levels of lncRNA-5657 and proinflammatory cytokines in BALF of patients and rats as well as in rat macrophages were measured. Changes in histopathologic score, lung wet/dry weight ratio, and spns2 expression in macrophages were examined. The relationship between lncRNA-5657 and its potential target gene spns2 was validated using a dual-luciferase reporter assay.

Results: The lncRNA expression profile of LPS-stimulated macrophages demonstrated that lncRNA-5657 showed the greatest fold-change. The BALF cells of patients with sepsis-induced ARDS and the lung tissue of rats with CLP-induced sepsis had significantly increased lncRNA-5657 levels. LncRNA-5657 silencing alleviated CLP-induced lung inflammation in rats. In NR8383 cells, lncRNA-5657 overexpression enhanced, whereas lncRNA-5657 silencing attenuated the expression of proinflammatory cytokines and spns2. A dual-luciferase reporter assay showed that lncRNA-5657 interacted with the promoter of the spns2 gene. Spns2 silencing alleviated LPS-induced inflammatory response and blocked the proinflammatory function of lncRNA-5657 in alveolar macrophages.

Conclusion: LncRNA-5657 is closely associated with sepsis-induce lung injury. In vitro and in vivo data demonstrated that LncRNA-5657 silencing alleviates sepsis-induced lung injury by inhibiting lung inflammatory response via suppressing spns2 expression.

Keywords: Acute respiratory distress syndrome; Alveolar macrophages; Inflammation; Long noncoding RNA; Sepsis.

MeSH terms

  • Acute Lung Injury / etiology
  • Acute Lung Injury / metabolism*
  • Acute Lung Injury / pathology
  • Animals
  • Anion Transport Proteins / metabolism*
  • Binding Sites
  • Bronchoalveolar Lavage Fluid / chemistry
  • Cecum / injuries
  • Cecum / surgery
  • Cell Line
  • Cytokines / metabolism
  • Disease Models, Animal
  • Fatty Acid Transport Proteins / metabolism*
  • Gene Knockdown Techniques
  • Gene Silencing
  • Humans
  • Inflammation / complications
  • Inflammation / metabolism
  • Lipopolysaccharides / adverse effects
  • Lipopolysaccharides / genetics
  • Macrophages, Alveolar / metabolism
  • RNA, Long Noncoding / biosynthesis*
  • RNA, Long Noncoding / genetics
  • Rats, Sprague-Dawley
  • Respiratory Distress Syndrome / etiology
  • Respiratory Distress Syndrome / genetics*
  • Respiratory Distress Syndrome / metabolism
  • Sepsis / complications
  • Sepsis / physiopathology*
  • Up-Regulation

Substances

  • Anion Transport Proteins
  • Cytokines
  • Fatty Acid Transport Proteins
  • Lipopolysaccharides
  • RNA, Long Noncoding
  • Spns2 protein, human
  • Spns2 protein, rat