FTY720 Inhibits MPP+-Induced Microglial Activation by Affecting NLRP3 Inflammasome Activation

J Neuroimmune Pharmacol. 2019 Sep;14(3):478-492. doi: 10.1007/s11481-019-09843-4. Epub 2019 May 8.

Abstract

Parkinson's disease (PD) is characterized by the degeneration of dopaminergic neurons and excessive microglial activation in the substantia nigra pars compacta (SNpc). In the present study, we aimed to demonstrate the therapeutic effectiveness of the potent sphingosine-1-phosphate receptor antagonist fingolimod (FTY720) in an animal model of PD induced by 1-methyl-4-phenyl-1,2,3,6-tetrahydropyridine (MPTP), and to identify the potential mechanisms underlying these therapeutic effects. C57BL/6J mice were orally administered FTY720 before subcutaneous injection of MPTP. Open-field and rotarod tests were performed to determine the therapeutic effect of FTY720. The damage to dopaminergic neurons and the production of monoamine neurotransmitters were assessed using immunohistochemistry, high-performance liquid chromatography, and flow cytometry. Immunofluorescence (CD68- positive) and enzyme-linked immunosorbent assay were used to analyze the activation of microglia, and the levels of activated signaling molecules were measured using Western blotting. Our findings indicated that FTY720 significantly attenuated MPTP-induced behavioral deficits, reduced the loss of dopaminergic neurons, and increased dopamine release. FTY720 directly inhibited MPTP-induced microglial activation in the SNpc, suppressed the production of interleukin (IL)-6, IL-1β, and tumor necrosis factor-α in BV-2 microglial cells treated with 1-methyl-4-phenylpyridinium (MPP+), and subsequently decreased apoptosis in SH-SY5Y neuroblastoma cells. Moreover, in MPP+-treated BV-2 cells and primary microglia, FTY720 treatment significantly attenuated the increases in the phosphorylation of PI3K/AKT/GSK-3β, reduced ROS generation and p65 activation, and also inhibited the activation of NLRP3 inflammasome and caspase-1. In conclusion, FTY720 may reduce PD progression by inhibiting NLRP3 inflammasome activation via its effects on ROS generation and p65 activation in microglia. These findings provide novel insights into the mechanisms underlying the therapeutic effects of FTY720, suggesting its potential as a novel therapeutic strategy against PD. Graphical Abstract FTY720 may reduce ROS production by inhibiting the PI3K/AKT/GSK-3β signaling pathway, while at the same time reducing p65 phosphorylation, thus decreasing NLRP3 inflammasome activation through these two pathways, ultimately reducing microglia activation-induced neuronal damage.

Keywords: Fingolimod (FTY720); Microglia; NLRP3 inflammasome; Parkinson’s disease; Reactive oxygen species (ROS).

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Antiparkinson Agents / pharmacology*
  • Apoptosis / drug effects
  • Cell Line
  • Corpus Striatum / chemistry
  • Corpus Striatum / drug effects
  • Cytokines / biosynthesis
  • Dopaminergic Neurons / drug effects
  • Dopaminergic Neurons / pathology
  • Drug Evaluation, Preclinical
  • Exploratory Behavior / drug effects
  • Fingolimod Hydrochloride / pharmacology*
  • Inflammasomes / drug effects*
  • Inflammasomes / metabolism
  • MPTP Poisoning / drug therapy
  • MPTP Poisoning / immunology
  • Male
  • Mice
  • Mice, Inbred C57BL
  • Microglia / drug effects*
  • Microglia / metabolism
  • Mitochondria / drug effects
  • NLR Family, Pyrin Domain-Containing 3 Protein / metabolism
  • Neuroprotective Agents / pharmacology*
  • Parkinsonian Disorders / drug therapy*
  • Parkinsonian Disorders / immunology
  • Pars Compacta / chemistry
  • Pars Compacta / drug effects
  • Pars Compacta / pathology
  • Reactive Oxygen Species
  • Rotarod Performance Test
  • Signal Transduction / drug effects

Substances

  • Antiparkinson Agents
  • Cytokines
  • Inflammasomes
  • NLR Family, Pyrin Domain-Containing 3 Protein
  • Neuroprotective Agents
  • Nlrp3 protein, mouse
  • Reactive Oxygen Species
  • Fingolimod Hydrochloride