The Neuroprotective Effects of Necrostatin-1 on Subarachnoid Hemorrhage in Rats Are Possibly Mediated by Preventing Blood–Brain Barrier Disruption and RIP3-Mediated Necroptosis

Abstract

Despite the substantial efforts to elucidate the role of early brain injury in subarachnoid hemorrhage (SAH), an effective pharmaceutical therapy for patients with SAH continues to be unavailable. This study aims to reveal the role of necroptosis after SAH, and explore whether the disruption of the blood–brain barrier (BBB) and RIP3-mediated necroptosis following SAH in a rat SAH model are altered by necrostatin-1 via its selective inhibition of receptor-interacting protein kinase 1 (RIP1). Sixty-five rats were used in the experiments. The SAH model was established using endovascular perforation. Necrostatin-1 was intracerebroventricularly injected 1 h before SAH induction. The neuroprotective effects of necrostatin-1 were evaluated with multiple methods such as magnetic resonance imaging (MRI) scanning, immunohistochemistry, propidium iodide (PI) labeling, and western blotting. Pretreatment with necrostatin-1 attenuated brain swelling and reduced the lesion volume on T2 sequence and ventricular volume on MRI 72 h after SAH induction. Albumin leakage and the degradation of tight junction proteins were also ameliorated by necrostatin-1 administration. In addition, necrostatin-1 decreased the number of PI-positive cells in the basal cortex, reduced the levels of the RIP3 and MLKL proteins, and inhibited the production of the pro-inflammatory cytokines IL-1β, IL-6, and TNF-α. Based on the findings from the present study, the selective RIP1 inhibitor necrostatin-1 functioned as a neuroprotective agent after SAH by attenuating brain swelling and BBB disruption. Moreover, the necrostatin-1 pretreatment prevented SAH-induced necroptosis by suppressing the activity of the RIP3/MLKL signaling pathway. These results will provide insights into new drugs and pharmacological targets to manage SAH, which are worth further study.

Bisphenol A induces coronary endothelial cell necroptosis by activating RIP3/CamKII dependent pathway

“…Accordingly, the treatment of mice with BPA and the necroptosis inhibitor NEC-1 reversed cardiotoxic effects of BPA producing fewer hemorrhages and less inflammation. In support of our results, Chen J. et al showed that NEC-1 prevented the disruption of the brain-blood barrier reducing brain hemorrhages 75 . However, NEC-1 only induced a moderate improvement of cardiac function so the possibility of a direct effect of BPA on myocardium by a different pathway cannot be completely ruled out.…”

Necrostatin-1 and necroptosis inhibition: Pathophysiology and therapeutic implications

“…Apart from I/R injury, the protective effect of Nec-1 has been observed in other disease models like brain hemorrhage and traumatic injury, which is related to I/R injury or I/R disease mechanism. In animal models of intracerebral/subarachnoid hemorrhage (ICH/SAH), application of Nec-1 attenuates cell death, reduces hematoma volume and improves neurological outcomes [ 37 , 55 , 136 , 137 ]. Besides, both pre-treatment and post-treatment of Nec-1 exerts protection against brain hemorrhage [ 54 , 55 ], indicating a broad therapeutic window.…”

“…In animal models of intracerebral/subarachnoid hemorrhage (ICH/SAH), application of Nec-1 attenuates cell death, reduces hematoma volume and improves neurological outcomes [ 37 , 55 , 136 , 137 ]. Besides, both pre-treatment and post-treatment of Nec-1 exerts protection against brain hemorrhage [ 54 , 55 ], indicating a broad therapeutic window. The underlying molecular mechanism is still not definite, but it involves oxidative stress and inflammation: for example, Nec-1 alleviates glutathione (GSH) depletion in hemin-induced cell death and inhibits NLRP3 inflammasome activation; the attenuated oxidation and inflammation ameliorates brain swelling and blood-brain barrier disruption, finally inhibits the formation of brain edema [ 54 , 55 , 138 ].…”

“…Besides, both pre-treatment and post-treatment of Nec-1 exerts protection against brain hemorrhage [ 54 , 55 ], indicating a broad therapeutic window. The underlying molecular mechanism is still not definite, but it involves oxidative stress and inflammation: for example, Nec-1 alleviates glutathione (GSH) depletion in hemin-induced cell death and inhibits NLRP3 inflammasome activation; the attenuated oxidation and inflammation ameliorates brain swelling and blood-brain barrier disruption, finally inhibits the formation of brain edema [ 54 , 55 , 138 ]. In researches of traumatic brain injury (TBI) and spinal cord injury (SCI), necroptosis plays a significant role in neural cell death and participates in both primary and secondary injury [ 47 , 48 ].…”

Dose- and Time-Dependent Cytotoxicity of Carteolol in Corneal Endothelial Cells and the Underlying Mechanisms

“…Necrostatin-1 is a small-molecule inhibitor of necroptosis, which is characterized by its selective inhibitory impact on RIPK1 (Degterev et al, 2005(Degterev et al, , 2008. Numerous studies have recently discovered the neuroprotective effects of Nec-1, whereas the addition of Nec-1 significantly decreased the related RIPK/MLKL protein levels in necroptosis (Chen et al, 2019). Our results showed that Nec-1 administration significantly dampened the increased expression of RIPK1, RIPK3, MLKL, and pMLKL.…”