Necrostatin-1 (Nec-1) serves as a potent inhibitor of necroptosis, capable of crossing the blood-brain barrier, with an effective concentration (EC50) of 490 nM in Jurkat cells. It specifically inhibits RIP1 kinase activity (EC50=182 nM) and also functions as an inhibitor of IDO[1].
体内研究
Additionally, Nec-1 influences tubular dilation and the dynamics of peritubular capillary dilation following RCM exposure. Pre-treatment with a single intraperitoneal dose of Necrostatin-1 (1.65 mg/kg body weight) 15 minutes prior to RCM exposure obstructs the normalization of conditions within the observation timeframe[2].
体外研究
Nec-1 effectively protects L929 cells from TNFα-triggered necrotic death without the need for additional caspase inhibition[1].
Moreover, Nec-1 has been shown to block the expansion of peritubular capillaries induced by radiocontrast media (RCM), highlighting a potentially new function for the RIP1 kinase domain beyond cell death in managing microvascular blood flow and the pathophysiology of contrast-induced acute kidney injury (CIAKI)[2].
At a concentration of 30 μM, Nec-1 increases survival rates in cardiomyocyte progenitor cells (CMPCs) by preventing necrotic cell death[4].
作用机制
Necrostatin 1 is a small molecular that specifically and potently inhibits cell death caused by the stimulation of death-domain receptors in the presence of caspase inhibition. The inhibition of Necrostatin 1 on RIP1 activity was through the T-loop segment.
Necrostatin-1 细胞实验
Cell Line
Concentration
Treated Time
Description
References
HEK293T cells
10 μM
12 hours
Evaluation of the role of Cys191 in GSDMD-NT pore formation
Nat Immunol. 2020 Jul;21(7):736-745
BMDMs
50 µM
3 h
Inhibition of necroptosis, reducing hypoxia-induced cell death
J Exp Med. 2021 Sep 6;218(9):e20200667.
THP-1 cells
30 µM
1 hour
To evaluate the inhibitory effect of Necrostatin-1 on necroptosis in THP-1 cells. The results showed that Necrostatin-1 completely inhibited HMGB1 release, supporting the major contribution of necrotic cell death in VEXAS.
Nat Commun. 2024 Jan 30;15(1):910.
astrocytes
30 μM
4 h
To evaluate the effect of RIPK1 kinase activity on inflammatory gene expression in astrocytes. Results showed that Nec-1s inhibited RIPK1 kinase activity and reduced inflammatory gene expression.
Cell Rep. 2021 May 11;35(6):109112.
microglia
30 μM
2 h
To evaluate the effect of RIPK1 kinase activity on inflammation and cell death in microglia. Results showed that Nec-1s inhibited RIPK1 kinase activity and reduced cell death.
Cell Rep. 2021 May 11;35(6):109112.
Necrostatin-1 动物实验
Species
Animal Model
Administration
Dosage
Frequency
Description
References
Rats
Middle cerebral artery occlusion (MCAO) model
Intracerebroventricular injection
20 mM
Single injection, observed for 72 hours
Nec-1 treatment significantly reduced ischemia-induced brain damage and neurological deficits, inhibited RIPK1 phosphorylation and its downstream signaling pathways.
Aging Dis. 2019 Aug 1;10(4):807-817
Mice
Experimental autoimmune encephalomyelitis (EAE) model
Oral
30 mg/kg and 60 mg/kg
Twice daily, continuous treatment
To evaluate the therapeutic effect of RIPK1 kinase inhibitor in the EAE model. Results showed that RIPK1 kinase inhibitor significantly alleviated disease symptoms.
Cell Rep. 2021 May 11;35(6):109112.
Mice
Myocardial infarction model
Subcutaneous injection
5 mg/kg
Once daily, continued until the end of the experiment
Inhibition of necroptosis, improving survival and cardiac function after myocardial infarction
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