Bumetanide enhances phenobarbital efficacy in a neonatal seizure model

Abstract

Objectives: High levels of expression of the Na+‐K+‐2Cl− (NKCC1) cotransporter in immature neurons cause the accumulation of intracellular chloride and, therefore, a depolarized Cl− equilibrium potential (E Cl). This results in the outward flux of Cl− through GABA A channels, the opposite direction compared with mature neurons, in which GABA A receptor activation is inhibitory because Cl− flows into the cell. This outward flow of Cl− in neonatal neurons is excitatory and contributes to a greater seizure propensity and poor electroencephalographic response to GABAergic anticonvulsants such as phenobarbital and benzodiazepines. Blocking the NKCC1 transporter with bumetanide prevents outward Cl− flux and causes a more negative GABA equilibrium potential (E GABA) in immature neurons. We therefore tested whether bumetanide enhances the anticonvulsant action of phenobarbital in the neonatal brain Methods: Recurrent seizures were induced in the intact hippocampal preparation in vitro by continuous 5‐hour exposure to low‐Mg 2+ solution. The anticonvulsant efficacy of phenobarbital, bumetanide, and the combination of these drugs was studied Results: Phenobarbital failed to abolish or depress recurrent seizures in 70% of hippocampi. In contrast, phenobarbital in combination with bumetanide abolished seizures in 70% of hippocampi and significantly reduced the frequency, duration, and power of seizures in the remaining 30% Interpretation: Thus, alteration of Cl− transport by bumetanide enables the anticonvulsant action of phenobarbital in immature brain. This is a mechanistic demonstration of rational anticonvulsant polypharmacy. The combination of these agents may comprise an effective therapy for early‐life seizures. Ann Neurol 2007