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

Pei Y. Ng; Kathryn A. McIntosh; Gillian Hargrave; Ka H. Ho; Andrew Paul; Robin Plevin

doi:10.1016/j.cellsig.2018.07.016

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

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

Strathclyde Institute Of Pharmacy And Biomedical Sciences

Research output: Contribution to journal › Article

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.

Language	English
Pages	59-71
Number of pages	13
Journal	Cellular Signalling
Volume	51
Early online date	2 Aug 2018
DOIs	10.1016/j.cellsig.2018.07.016
Publication status	Published - 30 Nov 2018

Fingerprint

Purinergic Receptors

JNK Mitogen-Activated Protein Kinases

Endothelial Cells

Cytokines

Adenine Nucleotides

Interleukin-1beta

Anisomycin

Pharmacology

Adenosine A2A Receptors

Adenosine A1 Receptors

E-Selectin

Sorbitol

Protein Kinase Inhibitors

Prostaglandin-Endoperoxide Synthases

Cyclic AMP-Dependent Protein Kinases

Research

Endothelium

Phosphotransferases

Phosphorylation

Keywords

G-protein coupled receptors
inflammation
purinergic receptors
C-Jun N-terminal kinase
interleukin-1β

Cite this

Ng, P. Y., McIntosh, K. A., Hargrave, G., Ho, K. H., Paul, A., & Plevin, R. (2018). Inhibition of cytokine-mediated JNK signalling by purinergic P2Y11 receptors, a novel protective mechanism in endothelial cells. Cellular Signalling, 51, 59-71. https://doi.org/10.1016/j.cellsig.2018.07.016

Ng, Pei Y. ; 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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Ng, PY, McIntosh, KA, Hargrave, G, Ho, KH, Paul, A & Plevin, R 2018, 'Inhibition of cytokine-mediated JNK signalling by purinergic P2Y11 receptors, a novel protective mechanism in endothelial cells' Cellular Signalling, vol. 51, pp. 59-71. https://doi.org/10.1016/j.cellsig.2018.07.016

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

In: Cellular Signalling, Vol. 51, 30.11.2018, p. 59-71.

Research output: Contribution to journal › Article

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T1 - Inhibition of cytokine-mediated JNK signalling by purinergic P2Y11 receptors, a novel protective mechanism in endothelial cells

AU - Ng, Pei Y.

AU - McIntosh, Kathryn A.

AU - Hargrave, Gillian

AU - Ho, Ka H.

AU - Paul, Andrew

AU - Plevin, Robin

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N2 - 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.

AB - 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.

KW - G-protein coupled receptors

KW - inflammation

KW - purinergic receptors

KW - C-Jun N-terminal kinase

KW - interleukin-1β

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