Inhibition of cytokine-mediated JNK signalling by purinergic P2Y11 receptors, a novel protective mechanism in endothelial cells

Pei Yuen Ng, Kathryn A. McIntosh, Gillian Hargrave, Ka H. Ho, Andrew Paul, Robin Plevin

Research output: Contribution to journalArticle

1 Citation (Scopus)

Abstract

Previous research from our laboratory has demonstrated a novel phenomenon whereby GPCRs play a role in inhibiting cytokine-mediated c-Jun N-terminal kinase (JNK) signalling. So far this novel phenomenon seems to have been vastly overlooked, with little research in the area. Therefore, in this study we explored this further; by assessing the potential of P2YRs to mediate inhibition of cytokine-mediated JNK signalling and related functional outcomes in human endothelial cells. We utilised primary endothelial cells, and employed the use of endogenous activators of P2YRs and well characterised pharmacological inhibitors, to assess signalling parameters mediated by P2YRs, Interleukin-1β (IL-1β), TNFα and JNK. Activation of P2YRs with adenosine tri-phosphate (ATP) resulted in a time- and concentration-dependent inhibition of IL-1β-mediated phosphorylation of JNK and associated kinase activity. The effect was specific for cytokine-mediated JNK signalling, as ATP was without effect on JNK induced by other non-specific activators (e.g. sorbitol, anisomycin), nor effective against other MAPK pathways such as p38 and the canonical NFκB cascade. Pharmacological studies demonstrated a role for the P2Y11 receptor in mediating this effect, but not the P2Y1 nor the adenosine receptors (A1, A2A, A2B & A3). The novel Gαq/11 inhibitor YM254890 and a protein kinase A (PKA) inhibitor H89 both partially reversed ATP-mediated inhibition of IL-1β-stimulated JNK indicating involvement of both Gαq/11 and Gαs mediated pathways. ATP also partially reversed IL-1β-mediated induction of cyclo‑oxygenase-2 (COX-2) and E-selectin. Collectively, these studies indicate the potential for activation of purinergic receptors to protect the endothelium from inflammatory driven JNK activation and may be a new target for inflammatory disease therapy.

Original languageEnglish
Pages (from-to)59-71
Number of pages13
JournalCellular Signalling
Volume51
DOIs
Publication statusPublished - 1 Nov 2018

Fingerprint

Purinergic Receptors
JNK Mitogen-Activated Protein Kinases
Endothelial Cells
Cytokines
Adenine Nucleotides
Interleukin-1
Anisomycin
Pharmacology
Adenosine A2A Receptors
Adenosine A1 Receptors
E-Selectin
Sorbitol
Cyclooxygenase 2
Protein Kinase Inhibitors
Cyclic AMP-Dependent Protein Kinases
Research
Endothelium
Phosphotransferases
Phosphorylation

Keywords

  • C-Jun N-terminal kinase
  • G-protein coupled receptors
  • H-89 (PubChem CID: 449241)
  • Inflammation
  • Interleukin-1β
  • MRS2179 (PubChem CID: 24867852)
  • NF340 (PubChem CID: 73755007)
  • Purinergic receptors
  • YM-254890 (PubChem CID: 9919454)
  • ZM241385 (PubChem CID: 176407)

ASJC Scopus subject areas

  • Cell Biology

Cite this

Inhibition of cytokine-mediated JNK signalling by purinergic P2Y11 receptors, a novel protective mechanism in endothelial cells. / Ng, Pei Yuen; McIntosh, Kathryn A.; Hargrave, Gillian; Ho, Ka H.; Paul, Andrew; Plevin, Robin.

In: Cellular Signalling, Vol. 51, 01.11.2018, p. 59-71.

Research output: Contribution to journalArticle

Ng, Pei Yuen ; McIntosh, Kathryn A. ; Hargrave, Gillian ; Ho, Ka H. ; Paul, Andrew ; Plevin, Robin. / Inhibition of cytokine-mediated JNK signalling by purinergic P2Y11 receptors, a novel protective mechanism in endothelial cells. In: Cellular Signalling. 2018 ; Vol. 51. pp. 59-71.
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