Cross-talk between the unfolded protein response and nuclear factor-κB signalling pathways regulates cytokine-mediated beta cell death in MIN6 cells and isolated mouse islets

Diabetologia. 2012 Nov;55(11):2999-3009. doi: 10.1007/s00125-012-2657-3. Epub 2012 Jul 28.

Abstract

Aims/hypothesis: Pancreatic beta cell destruction in type 1 diabetes may be mediated by cytokines such as IL-1β, IFN-γ and TNF-α. Endoplasmic reticulum (ER) stress and nuclear factor-κB (NFκB) signalling are activated by cytokines, but their significance in beta cells remains unclear. Here, we investigated the role of cytokine-induced ER stress and NFκB signalling in beta cell destruction.

Methods: Isolated mouse islets and MIN6 beta cells were incubated with IL-1β, IFN-γ and TNF-α. The chemical chaperone 4-phenylbutyric acid (PBA) was used to inhibit ER stress. Protein production and gene expression were assessed by western blot and real-time RT-PCR.

Results: We found in beta cells that inhibition of cytokine-induced ER stress with PBA unexpectedly potentiated cell death and NFκB-regulated gene expression. These responses were dependent on NFκB activation and were associated with a prolonged decrease in the inhibitor of κB-α (IκBα) protein, resulting from increased IκBα protein degradation. Cytokine-mediated NFκB-regulated gene expression was also potentiated after pre-induction of ER stress with thapsigargin, but not tunicamycin. Both PBA and thapsigargin treatments led to preferential upregulation of ER degradation genes over ER-resident chaperones as part of the adaptive unfolded protein response (UPR). In contrast, tunicamycin activated a balanced adaptive UPR in association with the maintenance of Xbp1 splicing.

Conclusions/interpretation: These data suggest a novel mechanism by which cytokine-mediated ER stress interacts with NFκB signalling in beta cells, by regulating IκBα degradation. The cross-talk between the UPR and NFκB signalling pathways may be important in the regulation of cytokine-mediated beta cell death.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antineoplastic Agents / pharmacology
  • Cell Death / drug effects
  • Cell Death / physiology*
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cell Survival / physiology
  • Cytokines / metabolism*
  • Cytokines / pharmacology
  • Diabetes Mellitus, Type 1 / metabolism*
  • Diabetes Mellitus, Type 1 / pathology
  • Endoplasmic Reticulum Stress / drug effects
  • Endoplasmic Reticulum Stress / physiology
  • Enzyme Inhibitors / pharmacology
  • Insulin-Secreting Cells / drug effects
  • Insulin-Secreting Cells / metabolism*
  • Insulin-Secreting Cells / pathology
  • Interferon-gamma / metabolism
  • Interferon-gamma / pharmacology
  • Interleukin-1beta / metabolism
  • Interleukin-1beta / pharmacology
  • Mice
  • Mice, Inbred C57BL
  • NF-kappa B / metabolism*
  • Pancreatic Neoplasms
  • Phenylbutyrates / pharmacology
  • RNA, Small Interfering / pharmacology
  • Signal Transduction / drug effects
  • Signal Transduction / physiology
  • Thapsigargin / pharmacology
  • Transcription Factor CHOP / genetics
  • Transcription Factor CHOP / metabolism
  • Tumor Necrosis Factor-alpha / metabolism
  • Tumor Necrosis Factor-alpha / pharmacology
  • Tunicamycin / pharmacology
  • Unfolded Protein Response / physiology*

Substances

  • Antineoplastic Agents
  • Cytokines
  • Ddit3 protein, mouse
  • Enzyme Inhibitors
  • Interleukin-1beta
  • NF-kappa B
  • Phenylbutyrates
  • RNA, Small Interfering
  • Tumor Necrosis Factor-alpha
  • Tunicamycin
  • Transcription Factor CHOP
  • Thapsigargin
  • 4-phenylbutyric acid
  • Interferon-gamma