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The neuroprotective effect of ONO-1714 on NMDA-mediated cytotoxicity in vitro
Satoshi Shibuta1, Sriranganathan Varathan, Takashi Mashimo
1Department of Anesthesiology and Acute Critical Medicine, Graduate School of Medicine, Osaka University (D7), 2-2 Yamadaoka, Osaka 565-0871, Suita, Japan. shibuta@anes.med.osaka-u.ac.jp
Abstract:
We report the effects of a newly developed NOS inhibitor on the neurotoxicity induced by NMDA on cultured fetal rat cortical neurons. To date, three different isoforms of NOS have been characterized. It has been considered that both neuronal NOS and inducible NOS activities are detrimental to the ischemic brain, whereas endothelial NOS plays a prominent role in maintaining cerebral blood flow and prevents neuronal injury during ischemia. ONO-1714 is a newly developed competitive NOS inhibitor that has selective inhibitory potency for iNOS than eNOS. However, its effect on nNOS has not been investigated yet. In this study, we investigated the neuroprotective effect of ONO-1714 on NMDA-induced neurotoxicity in our established model of primary cultured cortical neurons of rat foetus. Cortical neurons (prepared from E16 rat foetuses) were used after 13-14 days in culture. The cells were exposed to 30 muM NMDA for 24 h in the culture. To evaluate the neuroprotective effects of NOS inhibitors, ONO-1714 and L-NAME, neurons were exposed to various concentrations of an NOS inhibitor with 30 muM NMDA. The NMDA induced neurotoxicity was significantly attenuated by ONO-1714 in all concentrations, but not in low to moderate concentrations of L-NAME. These findings demonstrate that the neuroprotective effect of ONO-1714 was more potent than L-NAME. Moreover, ONO-1714 has a strong inhibitory effect on nNOS and would be a powerful tool for the protection of neurons against cerebral ischemia.
Insights
A new nitric oxide synthase (NOS) inhibitor, ONO-1714, effectively protected cultured rat neurons from NMDA-induced neurotoxicity. This finding suggests ONO-1714 is a potent neuroprotective agent against conditions like cerebral ischemia.
Area of Science:
- Neuroscience
- Pharmacology
- Cell Biology
Background:
- Nitric oxide synthase (NOS) has three isoforms: neuronal NOS (nNOS), inducible NOS (iNOS), and endothelial NOS (eNOS).
- nNOS and iNOS are implicated in brain injury during ischemia, while eNOS is protective.
- The novel NOS inhibitor ONO-1714 selectively targets iNOS but its effect on nNOS is unknown.
Purpose of the Study:
- To investigate the neuroprotective effects of ONO-1714 against N-methyl-D-aspartate (NMDA)-induced neurotoxicity in primary cultured rat cortical neurons.
- To compare the efficacy of ONO-1714 with L-NAME, another NOS inhibitor, in this neurotoxicity model.
Main Methods:
- Primary cortical neurons were cultured from E16 rat fetuses.
- Neurons were exposed to 30 μM NMDA for 24 hours to induce toxicity.
- Neuroprotection was evaluated by treating neurons with varying concentrations of ONO-1714 or L-NAME alongside NMDA.
Main Results:
- ONO-1714 significantly attenuated NMDA-induced neurotoxicity across all tested concentrations.
- L-NAME showed neuroprotective effects only at low to moderate concentrations.
- ONO-1714 demonstrated more potent neuroprotection than L-NAME.
Conclusions:
- ONO-1714 exhibits potent neuroprotective effects against NMDA-induced neurotoxicity in cultured cortical neurons.
- The study indicates ONO-1714 has a strong inhibitory effect on nNOS.
- ONO-1714 represents a promising therapeutic tool for protecting neurons against cerebral ischemia.
