Table 2.

Animal studies on the treatment of stroke by targeting necroptosis.

Authors		Interventions		Animal Models		Results
Zille et al. 2017		Chemical inhibitors implicated in all known cell death pathways		Cultured neurons exposed to hemoglobin or hemin		Experimental intracerebral hemorrhage shares features of ferroptotic and necroptotic, but not caspase-dependent apoptosis or autophagy
Shen et al. 2017		Nec-1 Mutation of serine kinase phosphorylation site of RIP1		ICH in rats		Inhibition of necroptosis
Qu et al. 2017		Inhibition of the RIP3-MLKL or RIP3-CaMKIIdelta interaction		OGD/zVAD in oligodendrocytes HI in postnatal day 6 (P6) rats		Disrupted development of myelin was attenuated
Xu et al. 2017		Necroptosis, autophagy, and apoptosis inhibitor		Global cerebral I/R injury model in rats		Mitochondria are involved in the execution of programmed necrosis, and AIF is the mediating molecule
Qu et al. 2016		Inhibition of MLKL		OGD/zVAD in cortical neurons HI in rats		Attenuated neuronal death induced by OGD/zVAD and brain damage induced by HI
LaRocca et al. 2016		nec-1s Treatment with high levels of glucose		HI in mice		Resulted in increased infarct size, which can be prevented by nec-1s
Yin et al. 2015		Nec-1 pretreatment		Global cerebral I/R injury		Nec-1 pretreatment prevented hippocampal CA1 neuronal death and I/R induced changes in RIP3
Xuan et al. 2015		A Water-Soluble Extract from the Culture Medium of Ganoderma lucidum Mycelia		HI in type 2 diabetic KKAy mice		Reduced H/I-induced neurological deficits and brain infarction volume and suppressed superoxide production, neuronal cell death, and vacuolation in the ischemic penumbra
Wang et al. 2015		extracellular protons		MCAO in mice		Acid stimulation recruits RIP1 to the ASIC1a C-terminus, causing RIP1 phosphorylation and subsequent neuronal death
Su et al. 2015		Nec-1 pretreatment		ICH in mice		Nec-1 pretreatment improved neurological function, attenuated brain edema, reduced RIP1-RIP3 interaction and PI positive cell death and inhibited microglia activation
Askalan et al. 2015		For 90min in the mild-moderate HI or 180min in the severe HI.		HI in rats		Necroptosis was significantly higher in the peri-infarct of the severe HI lesion compared to the moderate HI lesion. In males, but not in females, apoptosis was higher in moderate compared to severe HI
Vieira et al. 2014		RIP3 KD or overexpression		OGD in primary cultures of hippocampal neurons		RIP3 KD abrogated the component of OGD-induced necrotic neuronal death while RIP3 overexpression exacerbated neuronal death following OGD
Liu et al. 2014		RIP3 deficiency in mice KD of CYLD or RIP1 or RIP3 or MLKL in HT-22 cells		Intracerebroventricular injection of TNF-alpha in mice TNF-alpha-induced toxicity of hippocampal neurons in HT-22 cells		RIP3 deficiency attenuates TNF-alpha-initiated loss of hippocampal neurons The cell death is suppressed by KD of CYLD or RIP1 or RIP3 or MLKL
King et al. 2014		Nec-1		ICH in mice		Nec-1 significantly reduced hematoma volume and BBB opening, attenuated edema development, and improved neurobehavioral outcomes
Chang et al. 2014		Nec-1		ICH in mice		Nec-1 suppressed apoptosis, autophagy, and necroptosis to exert these neuroprotective effects after ICH
Dai et al. 2013		Curcumin		Iron induced neurotoxicity in primary cortical neurons		Curcumin attenuated necroptosis in a dose-dependent manner and decreased expression of receptor interacting protein 1 in a dose- and time-dependent manner
Zhu et al. 2012		RIP3 deficiency		ICH in mice		Mice deficient in RIP3 had 50% less PI+ cells at 24 h. Permeable cells remained in the brain for at least 24 h with <10% spontaneous resealing
Chen et al. 2012		GA		OGD/zVAD in cultured primary neurons		GA protected against neuronal injury and decreased RIP1 protein level in a time- and concentration-dependent manner
Authors	Interventions		Animal Models		Results
Chavez-Valdez et al. 2012	Nec-1 immediately after HI		Neonatal HI in mice		Nec-1 immediately after HI, is strongly mitoprotective and prevents secondary energy failure by blocking early NO* accumulation, glutathione oxidation and attenuating mitochondrial dysfunction
Northington et al. 2011	Nec-1		Neonatal HI in mice		Necrostatin treatment attenuated necrotic cell death, HI-induced oxidative damage and markers of inflammation
Xu et al. 2010	Nec-1 HNG		OGD in cultured mouse primary cortical neurons MCAO in mice		Nec-1 or HNG alone had protective effects on OGD-induced cell death. Combined treatment with Nec-1 and HNG resulted in more neuroprotection than Nec-1 or HNG alone
Xu et al. 2010	PJ34 Nec-1		Glutamate-induced necroptosis in HT-22 cells		Nec-1 is not a direct PARP inhibitor and that its signaling target is located upstream of PARP
Li et al. 2008	Nec-1		NMDA-induced excitotoxicity in rat's cultured cortical neurons		Nec-1 inhibited NMDA-induced decrease of cell viability, attenuated NMDA-induced leakage of LDH and suppressed NMDA-induced elevation of intracellular Ca2+
Degterev et al. 2005	Nec-1		Delayed mouse ischemic brain injury		A specific and potent small-molecule inhibitor of necroptosis, necrostatin-1, blocks a critical step in necroptosis

Abbreviations: RIP1: Receptor-interacting protein 1; RIP3: Receptor-interacting protein 3; MLKL: Mixed lineage kinase domain-like; Nec-1: Necrostatin-1, RIP1 inhibitor; Nec-1s: Nec-1 derivatives; KD: knock-down; I/R: Ischemia/reperfusion; HNG: Gly(14)-humanin, apoptosis inhibitor; GA: Geldanamycin; PJ34: a potent and specific inhibitor of poly(ADP-ribose)-polymerase (PARP); PI: Propidium iodide; BBB: Blood-brain barrier; ICH: Collagenase-induced intracerebral hemorrhage; OGD/zVAD: Oxygen-glucose deprivation plus caspase inhibitor zVAD treatment; OGD: Oxygen-glucose deprivation; HI: Hypoxia-ischemia; MCAO: Middle cerebral artery occlusion; ASIC1a: Acid-sensing ion channel 1; ICH: Intracerebral hemorrhage; LDH: Lactate dehydrogenase; NMDA: N-methyl-D-aspartic acid.