title: Methazolamide and melatonin inhibit mitochondrial cytochrome C release and are neuroprotective in experimental models of ischemic injury.
abstract: OBJECTIVE
  The identification of a neuroprotective drug for stroke remains elusive. Given that mitochondria play a key role both in maintaining cellular energetic homeostasis and in triggering the activation of cell death pathways, we evaluated the efficacy of newly identified inhibitors of cytochrome c release in hypoxia/ischemia induced cell death. We demonstrate that methazolamide and melatonin are protective in cellular and in vivo models of neuronal hypoxia.
  
  
  METHODS
  The effects of methazolamide and melatonin were tested in oxygen/glucose deprivation-induced death of primary cerebrocortical neurons. Mitochondrial membrane potential, release of apoptogenic mitochondrial factors, pro-IL-1beta processing, and activation of caspase -1 and -3 were evaluated. Methazolamide and melatonin were also studied in a middle cerebral artery occlusion mouse model. Infarct volume, neurological function, and biochemical events were examined in the absence or presence of the 2 drugs.
  
  
  RESULTS
  Methazolamide and melatonin inhibit oxygen/glucose deprivation-induced cell death, loss of mitochondrial membrane potential, release of mitochondrial factors, pro-IL-1beta processing, and activation of caspase-1 and -3 in primary cerebrocortical neurons. Furthermore, they decrease infarct size and improve neurological scores after middle cerebral artery occlusion in mice.
  
  
  CONCLUSIONS
  We demonstrate that methazolamide and melatonin are neuroprotective against cerebral ischemia and provide evidence of the effectiveness of a mitochondrial-based drug screen in identifying neuroprotective drugs. Given the proven human safety of melatonin and methazolamide, and their ability to cross the blood-brain-barrier, these drugs are attractive as potential novel therapies for ischemic injury.
authors.affiliation: Department of Neurosurgery, Neuroapoptosis Laboratory, Brigham and Women's Hospital, Harvard Medical School, Boston, Massachusetts 02115, USA.
authors.forename: Xin
authors.initials: X
authors.lastname: Wang
authors.forename: Bryan E
authors.initials: BE
authors.lastname: Figueroa
authors.forename: Irina G
authors.initials: IG
authors.lastname: Stavrovskaya
authors.forename: Yi
authors.initials: Y
authors.lastname: Zhang
authors.forename: Ana C
authors.initials: AC
authors.lastname: Sirianni
authors.forename: Shan
authors.initials: S
authors.lastname: Zhu
authors.forename: Arthur L
authors.initials: AL
authors.lastname: Day
authors.forename: Bruce S
authors.initials: BS
authors.lastname: Kristal
authors.forename: Robert M
authors.initials: RM
authors.lastname: Friedlander
mesh_terms: D000818:Animals; D000975:Antioxidants / Q000494:pharmacology*; D015153:Blotting, Western; D002545:Brain Ischemia / Q000473:pathology* / Q000517:prevention & control*; D002257:Carbonic Anhydrase Inhibitors / Q000494:pharmacology*; D020170:Caspase 1 / Q000378:metabolism; D053148:Caspase 3 / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D045304:Cytochromes c / Q000378:metabolism*; D004789:Enzyme Activation / Q000187:drug effects; D020287:In Situ Nick-End Labeling; D053583:Interleukin-1beta / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008550:Melatonin / Q000494:pharmacology*; D008564:Membrane Potentials / Q000187:drug effects; D008704:Methazolamide / Q000494:pharmacology*; D051379:Mice; D008810:Mice, Inbred C57BL; D008928:Mitochondria / Q000187:drug effects / Q000201:enzymology*; D051336:Mitochondrial Membranes / Q000187:drug effects; D019636:Neurodegenerative Diseases / Q000473:pathology; D009474:Neurons / Q000473:pathology; D018696:Neuroprotective Agents
chemical_list: D000975:Antioxidants; D002257:Carbonic Anhydrase Inhibitors; D053583:Interleukin-1beta; D018696:Neuroprotective Agents; D045304:Cytochromes c; D007770:L-Lactate Dehydrogenase; D053148:Caspase 3; D020170:Caspase 1; D008550:Melatonin; D008704:Methazolamide