Inhibition of Neuronal Necroptosis Mediated by RIPK1 Provides Neuroprotective Effects on Hypoxia and Ischemia In Vitro and In Vivo

Abstract

Ischemic brain injury is a widespread pathological condition, the main components of which are a deficiency of oxygen and energy substrates. In recent years, a number of new forms of cell death, including necroptosis, have been described. In necroptosis, a cascade of interactions between the kinases RIPK1 and RIPK3 and the MLKL protein leads to the formation of a specialized death complex called the necrosome, which triggers MLKL-mediated destruction of the cell membrane and necroptotic cell death. Necroptosis probably plays an important role in the development of ischemia/reperfusion injury and can be considered as a potential target for finding methods to correct the disruption of neural networks in ischemic damage. In the present study, we demonstrated that blockade of RIPK1 kinase by Necrostatin-1 preserved the viability of cells in primary hippocampal cultures in an in vitro model of glucose deprivation. The effect of RIPK1 blockade on the bioelectrical and metabolic calcium activity of neuron-glial networks in vitro using calcium imaging and multi-electrode arrays was assessed for the first time. RIPK1 blockade was shown to partially preserve both calcium and bioelectric activity of neuron-glial networks under ischemic factors. However, it should be noted that RIPK1 blockade does not preserve the network parameters of the collective calcium dynamics of neuron-glial networks, despite the maintenance of network bioelectrical activity (the number of bursts and the number of spikes in the bursts). To confirm the data obtained in vitro, we studied the effect of RIPK1 blockade on the resistance of small laboratory animals to in vivo modeling of hypoxia and cerebral ischemia. The use of Necrostatin-1 increases the survival rate of C57BL mice in modeling both acute hypobaric hypoxia and ischemic brain damage.

PERK‐STING‐RIPK3 pathway facilitates cognitive impairment by inducing neuronal necroptosis in sepsis‐associated encephalopathy

“…23 Subsequently, the MLKL binds to the cell membrane, causing a hole in the cell membrane, thereby triggering necroptosis. 24 Accumulating evidence suggests that cognitive impairments are associated with Alzheimer disease (AD), 21,25,26 specifically neuronal loss in the hippocampal CA1 regions. 27 In our study, protein expression levels of p-RIPK3/RIPK3 and p-MLKL/MLKL were meaningfully increased in hippocampal tissue of SAE mice.…”

Unravelling Contributions of Astrocytic Connexin 43 to the Functional Activity of Brain Neuron–Glial Networks under Hypoxic State In Vitro

“…Only in the last decade, the attention of researchers was switched to the study of neural networks as the main functional units of the CNS responsible for the implementation of processing, storage and transmission of information, as well as for the implementation of higher cognitive functions, including memory, consciousness and emotional reactions [ 45 ]. This approach, using modern methods of neurobiology, makes it possible to analyze the tiny mechanisms of restructuring of the functional architectonics of neural networks under hypoxic damage, as well as the contribution of each element of the network to the development of the functional network response to the stress factor [ 46 , 47 ].…”

Necroptosis in CNS diseases: Focus on astrocytes

“…There were studies conducted on the potential therapeutic effect of Nec-1 in many diseases affecting all organ systems. Briefly, the neuroprotective effect of necrostatin in ischemic injuries (Liu et al, 2019a;Mitroshina et al, 2022), hydrocephalus (Liu et al, 2021a), mitochondrial dysfunction (Apaijai et al, 2021), intracranial hypertension (Qian et al, 2022) was revealed. Necrostatin reduces postoperative cognitive dysfunction (Yin et al, 2022), neonatal neurotoxicity (Xu et al, 2021b), and cardiotoxicity (Erdogmus Ozgen et al, 2022).…”