Calcium/calmodulin-dependent protein kinase IV signaling pathway is upregulated in experimental necrotizing enterocolitis

Pediatr Surg Int. 2020 Mar;36(3):271-277. doi: 10.1007/s00383-019-04615-w. Epub 2020 Jan 16.


Purpose: Activation of calcium/calmodulin-dependent protein kinase IV (CaMKIV) has been shown to increase intestinal injury and inhibit epithelial cell proliferation in dextran sulfate sodium (DSS)-induced colitis mice. However, the role of CaMKIV in necrotizing enterocolitis (NEC) is unknown. We aimed to study the expression and activation of CaMKIV in experimental NEC.

Methods: Following ethical approval, NEC (n = 10) was induced in C57BL/6 mouse pups by hypoxia, gavage hyperosmolar formula feeding and lipopolysaccharide from postnatal days P5 to 9. Breastfed pups served as control (n = 10). Mouse pups were sacrificed on P9 and the terminal ileum was harvested. Gene NEC injury was scored blindly by three independent investigators. CaMKIV, CREM and IL17 gene expression, and CaMKIV and pCaMKIV protein expression were assessed. The data were compared using Mann-Whitney U test. P < 0.05 was considered significant.

Results: Intestinal injury was induced in the NEC mice and confirmed by histological scoring and inflammatory cytokine IL6. CaMKIV and its downstream target genes of CREM and IL17 were significantly elevated in NEC mice relative to control. Similarly, phosphorylated-CaMKIV (pCaMKIV), the active form of CaMKIV, was more notably expressed in the NEC ileal tissue relative to control ileal tissue. Elevated pCaMKIV protein expression was also confirmed by western blot.

Conclusion: CaMKIV expression and activation are upregulated in experimental NEC suggesting a potential contributing factor in the pathogenesis of NEC.

Keywords: Calcium/calmodulin-dependent protein kinase IV (CaMKIV); IL17; Necrotizing enterocolitis (NEC); pCaMKIV.

Publication types

  • Randomized Controlled Trial, Veterinary

MeSH terms

  • Animals
  • Animals, Newborn
  • Calcium / metabolism
  • Calcium-Calmodulin-Dependent Protein Kinase Type 4 / biosynthesis
  • Calcium-Calmodulin-Dependent Protein Kinase Type 4 / genetics*
  • Cell Proliferation
  • Cytokines / metabolism
  • Disease Models, Animal
  • Enterocolitis, Necrotizing / metabolism*
  • Enterocolitis, Necrotizing / pathology
  • Gene Expression Regulation*
  • Ileum / metabolism*
  • Mice
  • Mice, Inbred C57BL
  • RNA / genetics*
  • Signal Transduction
  • Up-Regulation*


  • Cytokines
  • RNA
  • Calcium-Calmodulin-Dependent Protein Kinase Type 4
  • Calcium