TGR5 activation attenuates neuroinflammation via Pellino3 inhibition of caspase-8/NLRP3 after middle cerebral artery occlusion in rats

Hui Liang, Nathanael Matei, Devin W. McBride, Yang Xu, Zhenhua Zhou, Jiping Tang, Benyan Luo, John H. Zhang

Research output: Contribution to journalArticlepeer-review

Abstract

Background: Nucleotide-binding oligomerization domain-like receptor pyrin domain-containing protein 3 (NLRP3) plays an important role in mediating inflammatory responses during ischemic stroke. Bile acid receptor Takeda-G-protein-receptor-5 (TGR5) has been identified as an important component in regulating brain inflammatory responses. In this study, we investigated the mechanism of TGR5 in alleviating neuroinflammation after middle cerebral artery occlusion (MCAO). Methods: Sprague-Dawley rats were subjected to MCAO and TGR5 agonist INT777 was administered intranasally 1 h after MCAO. Small interfering RNAs (siRNA) targeting TGR5 and Pellino3 were administered through intracerebroventricular injection 48 h before MCAO. Infarct volumes and neurologic scores were evaluated, and ELISA, flow cytometry, immunofluorescence staining, immunoblotting, and co-immunoprecipitation were used for the evaluations. Results: Endogenous TGR5 and Pellino3 levels increased after MCAO. TGR5 activation by INT777 significantly decreased pro-inflammatory cytokine, cleaved caspase-8, and NLRP3 levels, thereby reducing brain infarctions; both short- and long-term neurobehavioral assessments showed improvements. Ischemic damage induced the interaction of TGR5 with Pellino3. Knockdown of either TGR5 or Pellino3 increased the accumulation of cleaved caspase-8 and NLRP3, aggravated cerebral impairments, and abolished the anti-inflammatory effects of INT777 after MCAO. Conclusions: TGR5 activation attenuated brain injury by inhibiting neuroinflammation after MCAO, which could be mediated by Pellino3 inhibition of caspase-8/NLRP3.

Original languageEnglish
Article number40
JournalJournal of Neuroinflammation
Volume18
Issue number1
DOIs
StatePublished - Feb 2 2021

ASJC Scopus Subject Areas

  • General Neuroscience
  • Immunology
  • Neurology
  • Cellular and Molecular Neuroscience

Keywords

  • Early brain injury
  • Inflammation
  • Middle cerebral artery occlusion
  • Neuroprotection
  • TGR5
  • RNA, Small Interfering/administration & dosage
  • NLR Family, Pyrin Domain-Containing 3 Protein/antagonists & inhibitors
  • Inflammation Mediators/antagonists & inhibitors
  • Injections, Intraventricular
  • Rats
  • Male
  • Administration, Intranasal
  • Rats, Sprague-Dawley
  • Caspase 8/metabolism
  • Brain/drug effects
  • Receptors, G-Protein-Coupled/agonists
  • Ubiquitin-Protein Ligases/antagonists & inhibitors
  • Animals
  • Cholic Acids/administration & dosage
  • Infarction, Middle Cerebral Artery/metabolism

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