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Neurochemical Research

, Volume 44, Issue 12, pp 2755–2764 | Cite as

PKC Mediates LPS-Induced IL-1β Expression and Participates in the Pro-inflammatory Effect of A2AR Under High Glutamate Concentrations in Mouse Microglia

  • Sheng-Yu Fu
  • Ren-Ping Xiong
  • Yan Peng
  • Zhuo-Hang Zhang
  • Xing Chen
  • Yan Zhao
  • Ya-Lei Ning
  • Nan Yang
  • Yuan-Guo Zhou
  • Ping LiEmail author
Original Paper
  • 65 Downloads

Abstract

Pathogens such as bacterial lipopolysaccharide (LPS) play an important role in promoting the production of the inflammatory cytokines interleukin-1 beta (IL-1β) and tumour necrosis factor-α (TNF-α) in response to infection or damage in microglia. However, whether different signalling pathways regulate these two inflammatory factors remains unclear. The protein kinase C (PKC) family is involved in the regulation of inflammation, and our previous research showed that the activation of the PKC pathway played a key role in the LPS-induced transformation of the adenosine A2A receptor (A2AR) from anti-inflammatory activity to pro-inflammatory activity under high glutamate concentrations. Therefore, in the current study, we investigated the role of PKC in the LPS-induced production of these inflammatory cytokines in mouse primary microglia. GF109203X, a specific PKC inhibitor, inhibited the LPS-induced expression of IL-1β messenger ribonucleic acid and intracellular protein in a dose-dependent manner. Moreover, 5 µM GF109203X prevented LPS-induced IL-1β expression but did not significantly affect LPS-induced TNF-α expression. PKC promoted IL-1β expression by regulating the activity of NF-κB but did not significantly impact the activity of ERK1/2. A2AR activation by CGS21680, an A2AR agonist, facilitated LPS-induced IL-1β expression through the PKC pathway at high glutamate concentrations but did not significantly affect LPS-induced TNF-α expression. Taken together, these results suggest a new direction for specific intervention with LPS-induced inflammatory factors in response to specific signalling pathways and provide a mechanism for A2AR targeting, especially after brain injury, to influence inflammation by interfering with A2AR.

Keywords

Inflammation Microglial LPS IL-1β expression TNF-α expression Protein kinase C 

Abbreviations

LPS

Lipopolysaccharide

IL-1β

Interleukin-1 beta

TNF-α

Tumour necrosis factor-α

TBI

Traumatic brain injury

NF-κB

Nuclear factor- kappa-B

A2AR

Adenosine A2A receptor

ERK1/2

Extracellular regulated protein kinases 1/2

MAPK

Mitogen-activated protein kinase

TLR4

Toll-like receptor 4

MyD88

Myeloid differentiation primary response 88

Iba-1

Ionized calcium-binding adaptor molecule 1

DABK

des-Arginine9-bradykinin

Notes

Acknowledgements

This work was supported by the Scientific Research Foundation for the Returned Overseas Chinese Scholars, State Education Ministry.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflicts of interest.

Supplementary material

11064_2019_2895_MOESM1_ESM.tif (7.4 mb)
Supplementary Fig. 1Inhibition of PKC abrogates changes in microglial morphology after LPS stimulation. Morphological changes with LPS stimulation in microglia after PKC inhibition (a). Nuclei are labelled with DAPI (blue), and Iba-1 is indicated by red fluorescence; scale bar = 50 µm. Evaluation of microglial cell sphericity (an index of cell activation) in four different groups.*P < 0.05 and **P < 0.01 compared with the LPS stimulation group; #P < 0.05 and ##P < 0.01 compared with the untreated control group. Supplementary material 1 (TIF 7591.1 kb)

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Copyright information

© Springer Science+Business Media, LLC, part of Springer Nature 2019

Authors and Affiliations

  1. 1.The Molecular Biology Center, State Key Laboratory of Trauma, Burn and Combined Injury, Research Institute of Surgery and Daping HospitalThird Military Medical UniversityChongqingChina

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