MicroRNA-483-3p Inhibitor Ameliorates Sepsis-Induced Intestinal Injury by Attenuating Cell Apoptosis and Cytotoxicity Via Regulating HIPK2

Mol Biotechnol. 2024 Feb;66(2):233-240. doi: 10.1007/s12033-023-00734-x. Epub 2023 Apr 19.

Abstract

Sepsis is a life-threatening syndrome that can result in multi-organ dysfunction. MicroRNA (miR)-483-3p was previously demonstrated to be upregulated in sepsis patients; however, its specific functions in sepsis-triggered intestinal injury remain unclarified. Human intestinal epithelial NCM460 cell line was stimulated with lipopolysaccharide (LPS) to mimic sepsis-induced intestinal injury in vitro. Terminal-deoxynucleotidyl transferase mediated nick end labeling (TUNEL) staining was utilized for examining cell apoptosis. Western blotting and real time quantitative polymerase chain reaction (RT-qPCR) were used for detecting molecular protein and RNA levels. LPS-induced cytotoxicity was determined by measuring concentrations of lactate dehydrogenase (LDH), diamine oxidase (DAO) and fatty acid binding protein 2 (FABP2). Luciferase reporter assay was utilized for verifying the interaction between miR-483-3p and homeodomain interacting protein kinase 2 (HIPK2). Inhibiting miR-483-3p alleviates LPS-triggered NCM460 cell apoptosis and cytotoxicity. miR-483-3p targeted HIPK2 in LPS-stimulated NCM460 cells. Knockdown of HIPK2 reversed the above effects mediated by miR-483-3p inhibitor. Inhibiting miR-483-3p ameliorates LPS-triggered apoptosis and cytotoxicity by targeting HIPK2.

Keywords: HIPK2; Intestinal injury; LPS; Sepsis; miR-483-3p.

MeSH terms

  • Apoptosis
  • Biological Assay
  • Carrier Proteins
  • Humans
  • Lipopolysaccharides / pharmacology
  • MicroRNAs* / genetics
  • Protein Serine-Threonine Kinases
  • Sepsis* / complications
  • Sepsis* / genetics

Substances

  • Lipopolysaccharides
  • MicroRNAs
  • HIPK2 protein, human
  • Carrier Proteins
  • Protein Serine-Threonine Kinases
  • MIRN483 microRNA, human