Neurosteroid inhibition of cell death.
Diverse gamma-aminobutyric acid (GABAA) receptor modulators exhibited novel cytoprotective effects and mechanisms of action in rabbit renal proximal tubules subjected to mitochondrial inhibition (antimycin A) or hypoxia. Cytoprotective potencies (50% effective concentration, EC50) were 0.3 nM allopregnanolone (AP) > 0.4 nM 17 alpha-OH-allopregnanolone (17 alpha-OH-AP) > 30 nM dehydroepiandrosterone sulfate (DHEAS) = 30 nM pregnenolone sulfate (PS) > 500 nM pregnenolone (PREG) > 30 microM muscimol > 10 mM GABA following antimycin A exposure. Maximal protection with AP and 17 alpha-OH-AP was 70%, whereas DHEAS, PS, PREG, and muscimol produced 100% cytoprotection. Experiments with AP, PS, and muscimol revealed the return of mitochondrial function and active Na+ transport following hypoxia/reoxygenation. Muscimol inhibited the antimycin A-induced influx of both extracellular Ca2+ and Cl- that occurs during the late phase of cell injury, whereas the neurosteroids only inhibited influx of Cl-. Radioligand binding studies with AP and PS did not reveal a specific binding site; however, structural requirements were observed for cytoprotective potency and efficacy. In conclusion, we suggest that the GABAA receptor modulators muscimol and neurosteroids are cytoprotective at different cellular sites in the late phase of cell injury; muscimol inhibits Ca2+ and subsequent Cl- influx, whereas the neurosteroids inhibit Cl- influx.
Waters
S L
SL
Department of Pharmacology and Toxicology, University of Arkansas for Medical Sciences, Little Rock 72205, USA.
Miller
G W
GW
Aleo
M D
MD
Schnellmann
R G
RG
D000470:Alkaloids / Q000494:pharmacology; D000818:Animals; D000968:Antimycin A / Q000494:pharmacology; D044182:Benzylisoquinolines; D002118:Calcium / Q000378:metabolism; D016923:Cell Death / Q000187:drug effects; D015687:Cell Hypoxia; D002478:Cells, Cultured; D019314:Dehydroepiandrosterone Sulfate / Q000494:pharmacology; D005260:Female; D007555:Isoxazoles / Q000494:pharmacology; D007687:Kidney Tubules, Proximal / Q000166:cytology* / Q000187:drug effects; D007700:Kinetics; D007770:L-Lactate Dehydrogenase; D008928:Mitochondria / Q000187:drug effects / Q000378:metabolism; D009118:Muscimol / Q000737:chemistry / Q000494:pharmacology; D010101:Oxygen Consumption / Q000187:drug effects; D011278:Pregnanes / Q000737:chemistry / Q000494:pharmacology*; D011817:Rabbits; D011963:Receptors, GABA-A / Q000187:drug effects / Q000502:physiology*; D005680:gamma-Aminobutyric Acid / Q000494:pharmacology*
D000470:Alkaloids; D044182:Benzylisoquinolines; D007555:Isoxazoles; D011278:Pregnanes; D011963:Receptors, GABA-A; D009118:Muscimol; D005680:gamma-Aminobutyric Acid; D019314:Dehydroepiandrosterone Sulfate; D000968:Antimycin A; C013024:hydrastine; D007770:L-Lactate Dehydrogenase; C015542:gaboxadol; D002118:Calcium