CircPRKCI relieves lipopolysaccharide-induced HK2 cell injury by upregulating the expression of miR-545 target gene ZEB2

Biofactors. 2020 May;46(3):475-486. doi: 10.1002/biof.1620. Epub 2020 Feb 27.

Abstract

The aim of this study was to investigate the possible influences of circPRKCI abnormal expression on lipopolysaccharide (LPS)-induced HK2 cell injury and its mechanism. The circPRKCI level was identified in serum samples from patients with urosepsis and healthy subjects, as well as LPS-treated HK2 cells by qRT-PCR. Cell viability, apoptosis, expression of proteins associated with apoptosis, and expression of pro-inflammatory cytokines in LPS-treated HK2 cells were measured. Effects of circPRKCI abnormal expression on LPS-induced HK2 cell injury were then evaluated. Afterward, the binding miRNA of circPRKCI and target gene of miRNA were identified, and the involvements of NF-kB pathway signaling pathway with the effects of circPRKCI were finally studied. CircPRKCI was significantly down-regulated in serum samples from patients with urosepsis and LPS-treated HK2 cells. LPS-induced decrease of cell viability, increase of cell apoptosis, as well as elevated productions of tumor necrosis factor (TNF)-α, interleukins (IL)-1β, IL-6, and IL-8 in HK2 cells were attenuated by overexpressed circPRKCI. In addition, circPRKCI negatively regulated the expression of miR-545, and miR-545 up-regulation reversed the inhibiting effects of circPRKCI overexpression on LPS-induced HK2 cell injury. Moreover, zinc finger E-box-binding homeobox 2 (ZEB2) was identified as a target gene of miR-545, and ZEB2 overexpression partly reversed the effects of miR-545 up-regulation on LPS-induced HK2 cell injury. Furthermore, NF-kB pathway was revealed to be associated to the effects of circPRKCI on LPS-induced HK2 cell injury. This research indicated that the highly expressed circPRKCI relieved inflammatory injury induced by LPS in HK2 cells by suppressing miR-545/ZEBs and depressing the briskness of NF-kB pathway.

Keywords: circular RNA; miR-545; sepsis-induced acute kidney injury; zinc finger E-box-binding homeobox 2.

MeSH terms

  • Adolescent
  • Adult
  • Aged
  • Cell Survival / genetics
  • Female
  • Hexokinase / blood
  • Hexokinase / genetics*
  • Humans
  • Inflammation / blood
  • Inflammation / genetics
  • Inflammation / prevention & control*
  • Isoenzymes / blood
  • Isoenzymes / genetics*
  • Lipopolysaccharides
  • Male
  • MicroRNAs / genetics*
  • Middle Aged
  • Protein Kinase C / blood
  • Protein Kinase C / genetics*
  • Signal Transduction / genetics
  • Up-Regulation / genetics
  • Young Adult
  • Zinc Finger E-box Binding Homeobox 2 / metabolism*

Substances

  • Isoenzymes
  • Lipopolysaccharides
  • MIR545 microRNA, human
  • MicroRNAs
  • ZEB2 protein, human
  • Zinc Finger E-box Binding Homeobox 2
  • HK2 protein, human
  • Hexokinase
  • Protein Kinase C
  • protein kinase C lambda