Dysfunction of mitochondria and deformed gap junctions in the heart of IL-18-deficient mice

Am J Physiol Heart Circ Physiol. 2016 Aug 1;311(2):H313-25. doi: 10.1152/ajpheart.00927.2015. Epub 2016 Jun 10.

Abstract

Interleukin-18 (IL-18) was discovered as an interferon-γ-inducing factor and has been regarded as a proinflammatory cytokine. However, IL-18 is ubiquitously expressed both in immune/inflammatory cells and in nonimmune cells, and its biological roles have not been sufficiently elucidated. Here, we demonstrate that IL-18-deficient [IL-18 knockout (KO)] mice have heart abnormalities that may be related to impaired autophagy. In endurance running tests, IL-18KO mice ran significantly shorter distances compared with wild-type (WT) mice. Echocardiographs indicated disability in the systolic and diastolic functions of the IL-18KO mouse heart. Immunostaining of connexin 43 showed heterogeneous localization of gap junctions in the lateral membranes of the IL-18KO cardiac myocytes. Western blotting analysis revealed decreased phosphorylated connexin 43 in the IL-18KO heart. Electron microscopy revealed unusual localization of intercalated disks, swollen or damaged mitochondria, and broad, indistinct Z-lines in the IL-18KO heart. In accordance with the morphological observation, mitochondrial respiratory function, including that of complexes I and IV, was impaired, and production of reactive oxygen species was augmented in IL-18KO hearts. Notably, levels of LC3-II were markedly lower in the IL-18KO hearts than in WT hearts. In the culture of cardiac myocytes of IL-18KO neonates, exogenous IL-18 upregulated LC3-II and increased the number of intact mitochondria with high mitochondrial membrane potential. These results indicated that IL-18 has roles apart from those as a proinflammatory cytokine in cardiac myocytes and suggested that IL-18 contributes to the homeostatic maintenance of mitochondrial function and gap-junction turnover in cardiac myocytes, possibly by upregulating autophagy.

Keywords: IL-18; cardiac myocytes; connexin 43; gap junction; mitochondrial autophagy.

MeSH terms

  • Animals
  • Autophagy / genetics*
  • Blotting, Western
  • Cell Membrane / metabolism
  • Cell Membrane / ultrastructure
  • Cells, Cultured
  • Connexin 43 / metabolism
  • Echocardiography
  • Electron Transport Complex I / metabolism
  • Electron Transport Complex IV / metabolism
  • Gap Junctions / ultrastructure*
  • Interleukin-18 / genetics*
  • Interleukin-18 / pharmacology
  • Male
  • Mice
  • Mice, Knockout
  • Microscopy, Electron
  • Microtubule-Associated Proteins / metabolism
  • Mitochondria, Heart / metabolism*
  • Myocytes, Cardiac / drug effects
  • Myocytes, Cardiac / metabolism*
  • Myocytes, Cardiac / ultrastructure
  • Phosphoproteins / metabolism
  • Phosphorylation
  • Physical Endurance
  • Reactive Oxygen Species / metabolism
  • Systole
  • Up-Regulation

Substances

  • Connexin 43
  • GJA1 protein, mouse
  • Interleukin-18
  • Map1lc3b protein, mouse
  • Microtubule-Associated Proteins
  • Phosphoproteins
  • Reactive Oxygen Species
  • Electron Transport Complex IV
  • Electron Transport Complex I