Inhibition of neuronal necroptosis mediated by RIP1/RIP3/MLKL provides neuroprotective effects on kaolin-induced hydrocephalus in mice

Cell Prolif. 2021 Sep;54(9):e13108. doi: 10.1111/cpr.13108. Epub 2021 Aug 9.

Abstract

Objectives: Necroptosis is widespread in neurodegenerative diseases. Here, we examined necroptosis in the hippocampus and cortex after hydrocephalus and found that a necroptosis pathway inhibitor alleviates necroptosis and provides neuroprotective effects.

Materials and methods: Hydrocephalus was induced in C57BL/6 mice by kaolin. Haematoxylin and eosin (HE), Nissl, PI and Fluoro-Jade B (FJB) staining were used for general observations. Phosphorylated receptor-interacting protein kinase 3 (p-RIP3) and phosphorylated mixed lineage kinase domain-like (p-MLKL) were measured by Western blotting and immunohistochemistry. Scanning electron microscopy (SEM) was used to observe ependymal cilia. Magnetic resonance imaging (MRI) and the Morris water maze (MWM) test were used to assess neurobehavioral changes. Immunofluorescence was used to detect microglial and astrocyte activation. Inflammatory cytokines were measured by Western blotting and RT-PCR.

Results: Obvious pathological changes appeared in the hippocampus and cortex after hydrocephalus, and expression of the necroptosis markers p-RIP3, p-MLKL and inflammatory cytokines increased. Necrostatin-1 (Nec-1) and GSK872 reduced necrotic cell death, attenuated p-RIP3 and p-MLKL levels, slightly improved neurobehaviours and inhibited microglial and astrocyte activation and inflammation.

Conclusions: RIP1/RIP3/MLKL mediates necroptosis in the cortex and hippocampus in a hydrocephalus mouse model, and Nec-1 and GSK872 have some neuroprotective effects.

Keywords: cortex; hippocampus; hydrocephalus; necroptosis; neuroinflammation.

MeSH terms

  • Animals
  • Cytokines / metabolism
  • Disease Models, Animal
  • GTPase-Activating Proteins / metabolism*
  • Hydrocephalus / chemically induced
  • Hydrocephalus / metabolism*
  • Imidazoles / metabolism
  • Indoles / metabolism
  • Inflammation / metabolism
  • Kaolin / pharmacology
  • Mice
  • Mice, Inbred C57BL
  • Necroptosis / drug effects
  • Necroptosis / physiology*
  • Neuroprotective Agents / metabolism*
  • Phosphorylation / drug effects
  • Protein Kinases / metabolism*
  • Receptor-Interacting Protein Serine-Threonine Kinases / metabolism*
  • Signal Transduction / drug effects

Substances

  • Cytokines
  • GTPase-Activating Proteins
  • Imidazoles
  • Indoles
  • Neuroprotective Agents
  • Ralbp1 protein, mouse
  • necrostatin-1
  • Kaolin
  • MLKL protein, mouse
  • Protein Kinases
  • Receptor-Interacting Protein Serine-Threonine Kinases
  • Ripk3 protein, mouse