RIPK3 activation promotes DAXX-dependent neuronal necroptosis after intracerebral hemorrhage in mice

Background: Necroptosis induced by receptor-interacting protein kinase 3 (RIPK3) is engaged in intracerebral hemorrhage (ICH) pathology. In this study, we explored the impact of RIPK3 activation on neuronal Necroptosis and the mechanism of the death domain-associated protein (DAXX)-mediated nuclear Necroptosis pathway after ICH.

Methods: Potential molecules linked to the progression of ICH were discovered using RNA sequencing. The level of DAXX was assessed by quantitative Real-Time PCR, ELISA, and western blotting. DAXX localization was determined by immunofluorescence and immunoprecipitation assays. The RIPK3 Inhibitor GSK872 and DAXX knockdown with shRNA-DAXX were used to examine the nuclear Necroptosis pathway associated with ICH. Neurobehavioral deficit assessments were performed.

Results: DAXX was increased in patients and mice after ICH. In an ICH mouse model, shRNA-DAXX reduced brain water content and alleviated neurologic impairments. GSK872 administration reduced the expression of DAXX. shRNA-DAXX inhibited the expression of p-MLKL. Immunofluorescence and immunoprecipitation assays showed that RIPK3 and AIF translocated into the nucleus and then bound with nuclear DAXX.

Conclusions: RIPK3 revitalization promoted neuronal Necroptosis in ICH mice, partially through the DAXX signaling pathway. RIPK3 and AIF interacted with nuclear DAXX to aggravate ICH injury.

death domain-associated protein; intracerebral hemorrhage; necroptosis; neurobehavioral deficits; receptor-interacting protein kinase 3.