Potentiation of hypoxic injury in cultured rabbit hepatocytes by the quinoxalinone anxiolytic, panadiplon.
The quinoxalinone anxiolytic, panadiplon, produces hepatic metabolic inhibition (mitochondrial impairment), microvesicular steatosis and centrilobular necrosis in rabbits. Metabolic inhibition occurs in cultured hepatocytes without cytotoxicity, suggesting that hepatic injury is influenced by additional factors. The present experiments were conducted to determine if metabolic inhibition by panadiplon predisposed hepatocytes to hypoxic injury. Injury (cell death) was evaluated by lactate dehydrogenase (LDH) release from cells; ATP and glycogen levels were also evaluated. Under hypoxic conditions, control cultures showed a 6.5-fold increase in LDH release compared to normoxic controls, with a coincident 80% decrease in ATP and 50% decrease in glycogen levels. Under normoxic conditions 10 microgram/ml panadiplon treatment for 48 h reduced ATP and glycogen levels by 40% but did not cause an increase in LDH leakage. Cells treated with panadiplon, then exposed to hypoxia conditions, showed a significant level of injury compared to normoxic control cultures, and a further reduction in ATP. No additional decrease in glycogen ws observed. In an attempt to prevent panadiplon-mediated injury, glycolytic substrates (dihydroxyacetone or pyruvate) were included during normoxic and hypoxic incubations. Both cotreatments reduced the level of LDH leakage produced by panadiplon during hypoxia. Cotreatment did not generally increase ATP or glycogen levels (compared to panadiplon treatment groups) during hypoxia, though individual experiments showed a slight increase in ATP levels. During normoxia both cotreatments with panadiplon resulted in significantly higher glycogen levels than in panadiplon cultures alone. These results suggest that cellular glycogen and subsequently ATP levels are reduced during panadiplon exposure, metabolically predisposing hepatocytes to hypoxic injury.
Bacon
J A
JA
Investigative Toxicology, Upjohn Laboratories, Kalamazoo, MI 49007, USA.
Cramer
C T
CT
Petrella
D K
DK
Sun
E L
EL
Ulrich
R G
RG
D000255:Adenosine Triphosphate / Q000378:metabolism; D000818:Animals; D014151:Anti-Anxiety Agents / Q000633:toxicity*; D016923:Cell Death; D015687:Cell Hypoxia; D002478:Cells, Cultured; D004098:Dihydroxyacetone / Q000494:pharmacology; D004357:Drug Synergism; D005260:Female; D006003:Glycogen / Q000378:metabolism; D007770:L-Lactate Dehydrogenase / Q000378:metabolism; D008099:Liver / Q000166:cytology / Q000187:drug effects* / Q000378:metabolism; D010069:Oxadiazoles / Q000633:toxicity*; D010100:Oxygen / Q000378:metabolism; D011773:Pyruvates / Q000494:pharmacology; D019289:Pyruvic Acid; D011810:Quinoxalines / Q000633:toxicity*; D011817:Rabbits
D014151:Anti-Anxiety Agents; D010069:Oxadiazoles; D011773:Pyruvates; D011810:Quinoxalines; D019289:Pyruvic Acid; D000255:Adenosine Triphosphate; D006003:Glycogen; D007770:L-Lactate Dehydrogenase; D004098:Dihydroxyacetone; D010100:Oxygen; C069443:panadiplon