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The antiapoptotic effect of remifentanil on the immature mouse brain: an ex vivo study
Fabien Tourrel1, Pamela Kwetieu de Lendeu, Lénaïg Abily-Donval
1From the *Laboratory of Microvascular Endothelium and Neonate Brain Lesions, Rouen Institute for Biomedical Research (IRIB), University of Rouen, Cedex, France; †Laboratory of Pharmaceutical Organic Chemistry, ULB, Brussels, Belgium; Departments of ‡Pharmacology, and §Anesthetics and Intensive Care, Rouen University Hospital, Rouen, France.
Background:
The use of remifentanil in a context of potential prematurity led us to explore ex vivo the opioid effects on the immature mouse brain. Remifentanil enhances medullary glutamatergic N-methyl-D-aspartate (NMDA) receptor activity. Furthermore, in neonatal mouse cortex, NMDA was previously shown to exert either excitotoxic or antiapoptotic effects depending on the cortical layers. With the use of a model of acute cultured brain slices, we evaluated the potential necrotic and apoptotic effects of remifentanil, alone or associated with its glycine vehicle (commercial preparation of remifentanil, C.P. remifentanil), on the immature brain.
Methods:
Cerebral slices from postnatal day 2 mice were treated up to 5 hours with the different compounds, incubated alone or in the presence of NMDA. The necrotic effect was studied by measuring lactate dehydrogenase activity and 7-Aminoactinomycin D labeling. Apoptotic death was evaluated by measurement of caspase-3 activity and cleaved caspase-3 protein levels, using Western blot and immunohistochemistry. Extrinsic and intrinsic apoptotic pathways were investigated by measuring caspase-8, caspase-9 activities, Bax protein levels, and mitochondrial integrity.
Results:
C.P. remifentanil was ineffective on necrotic death, whereas it significantly reduced caspase-3 activity and cortical cleaved caspase-3 levels. C.P. remifentanil inhibited cortical Bax protein expression, caspase-9 activity, and preserved mitochondrial integrity, whereas it had no effect on caspase-8 activity. Its action targeted the neocortex superficial layers, and it was reversed by the opioid receptors antagonist naloxone and the NMDA antagonist MK801. Remifentanil and glycine acted synergistically to inhibit apoptotic death. In addition, C.P. remifentanil enhanced the antiapoptotic effect of NMDA, whereas it did not improve NMDA excitotoxicity in brain slices.
Conclusion:
The present data indicate that at a supraclinical concentration C.P. remifentanil had no pronecrotic effect but exerted ex vivo antiapoptotic action on the immature mouse brain, involving the opioid and NMDA receptors, and the mitochondrial-dependent apoptotic pathway. Assessment of the impact of the antiapoptotic effect of remifentanil in in vivo neonatal mouse models of brain injury will also be essential to measure its consequences on the developing brain.
Insights
Commercial preparation of remifentanil (C.P. remifentanil) showed no harmful necrotic effects on immature mouse brains. Instead, it demonstrated an anti-apoptotic action, protecting against programmed cell death via opioid and NMDA receptors.
Area of Science:
- Neuroscience
- Pharmacology
- Developmental Biology
Background:
- Remifentanil use in premature infants prompted investigation into its effects on the immature brain.
- Remifentanil modulates N-methyl-D-aspartate (NMDA) receptor activity, which has dual effects (excitotoxic or anti-apoptotic) in the neonatal mouse cortex.
- The study aimed to assess the necrotic and apoptotic potential of remifentanil and its vehicle on immature brain slices.
Purpose of the Study:
- To evaluate the ex vivo effects of remifentanil, alone and with its glycine vehicle, on immature mouse brain slices.
- To determine if remifentanil induces necrotic or apoptotic cell death.
- To investigate the involvement of opioid and NMDA receptors in remifentanil's effects.
Main Methods:
- Cerebral slices from postnatal day 2 mice were cultured and treated with remifentanil, glycine, and NMDA.
- Necrotic effects were measured by lactate dehydrogenase activity and 7-Aminoactinomycin D labeling.
- Apoptotic effects were assessed via caspase-3 activity, Western blot, and immunohistochemistry, examining both extrinsic and intrinsic pathways.
Main Results:
- Commercial preparation of remifentanil (C.P. remifentanil) did not induce necrotic cell death.
- C.P. remifentanil significantly reduced apoptosis by inhibiting caspase-3 and caspase-9 activity and preserving mitochondrial integrity.
- Remifentanil's anti-apoptotic effects were mediated by opioid and NMDA receptors, acting synergistically with glycine and enhancing NMDA's anti-apoptotic properties without affecting excitotoxicity.
Conclusions:
- Ex vivo, C.P. remifentanil exhibits anti-apoptotic effects on the immature mouse brain at supraclinical concentrations, involving opioid, NMDA receptors, and the mitochondrial pathway.
- The study highlights the need for in vivo assessment of remifentanil's anti-apoptotic impact in neonatal brain injury models.
- Further research is crucial to understand the full consequences of remifentanil's effects on brain development.
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