Research Article| Volume 142, 109189, May 2023

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Bumetanide potentiates the anti-seizure and disease-modifying effects of midazolam in a noninvasive rat model of term birth asphyxia

  • Author Footnotes
    1 Present address: MSD Animal Health Innovation GmbH, Schwabenheim an der Selz, Germany.
    Björn Welzel
    1 Present address: MSD Animal Health Innovation GmbH, Schwabenheim an der Selz, Germany.
    Department of Pharmacology, Toxicology, and Pharmacy, University of Veterinary Medicine Hannover, Germany

    Center for Systems Neuroscience Hannover, Germany
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  • Author Footnotes
    2 Present address: Department of Neurosurgery, Hannover Medical School, Germany.
    Marie Johne
    2 Present address: Department of Neurosurgery, Hannover Medical School, Germany.
    Department of Pharmacology, Toxicology, and Pharmacy, University of Veterinary Medicine Hannover, Germany

    Center for Systems Neuroscience Hannover, Germany
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  • Wolfgang Löscher
    Corresponding author at: Department of Pharmacology, Toxicology and Pharmacy, University of Veterinary Medicine, Bünteweg 17, D-30559 Hannover, Germany.
    Department of Pharmacology, Toxicology, and Pharmacy, University of Veterinary Medicine Hannover, Germany

    Center for Systems Neuroscience Hannover, Germany
    Search for articles by this author
  • Author Footnotes
    1 Present address: MSD Animal Health Innovation GmbH, Schwabenheim an der Selz, Germany.
    2 Present address: Department of Neurosurgery, Hannover Medical School, Germany.


      • Birth asphyxia often causes hypoxic-ischemic encephalopathy, and neonatal seizures, and later-life adverse outcome.
      • In most cases, current therapies cannot prevent this outcome.
      • A rat model of birth asphyxia, neonatal seizures, and later-life adverse outcomes can be used to explore novel therapies.
      • The loop diuretic bumetanide enhances the anti-seizure and disease-modifying effects of midazolam in this model.
      • The mechanisms to explain that bumetanide potentiates midazolam but not phenobarbital remain to be determined.


      Birth asphyxia and the resulting hypoxic-ischemic encephalopathy (HIE) are highly associated with perinatal and neonatal death, neonatal seizures, and an adverse later-life outcome. Currently used drugs, including phenobarbital and midazolam, have limited efficacy to suppress neonatal seizures. There is a medical need to develop new therapies that not only suppress neonatal seizures but also prevent later-life consequences. We have previously shown that the loop diuretic bumetanide does not potentiate the effects of phenobarbital in a rat model of birth asphyxia. Here we compared the effects of bumetanide (0.3 or 10 mg/kg i.p.), midazolam (1 mg/kg i.p.), and a combination of bumetanide and midazolam on neonatal seizures and later-life outcomes in this model. While bumetanide at either dose was ineffective when administered alone, the higher dose of bumetanide markedly potentiated midazolam’s effect on neonatal seizures. Median bumetanide brain levels (0.47–0.53 µM) obtained with the higher dose were in the range known to inhibit the Na-K-Cl-cotransporter NKCC1 but it remains to be determined whether brain NKCC1 inhibition was underlying the potentiation of midazolam. When behavioral and cognitive alterations were examined over three months after asphyxia, treatment with the bumetanide/midazolam combination, but not with bumetanide or midazolam alone, prevented impairment of learning and memory. Furthermore, the combination prevented the loss of neurons in the dentate hilus and aberrant mossy fiber sprouting in the CA3a area of the hippocampus. The molecular mechanisms that explain that bumetanide potentiates midazolam but not phenobarbital in the rat model of birth asphyxia remain to be determined.



      CAI (carbonic anhydrase inhibitor), HIE (hypoxic-ischemic encephalopathy), NKCC (Na-K-Cl-cotransporter), P11 (postnatal day 11), RAWM (radial-arm water maze)
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