Validation of an in vitro model for assessment of androstenedione hepatotoxicity using the rat liver cell line clone-9.
Androstenedione, a naturally occurring steroid hormone, has been used to enhance athletic performance. Little is known, however, about its hepatotoxicity. Clone-9 cells, a non-transformed epithelial cell line that was originally isolated from normal liver of a 4-week old Sprague-Dawley rat, were used as an in vitro model to assess the hepatotoxic potential of androstenedione. The cultures were treated with androstenedione for 24 h at 37 degrees C in 5% CO(2) at concentrations of 0-100 microg ml(-1). After the treatment period, the cells and the culture supernatants were assayed for markers of cytotoxicity which included: release of liver enzymes, cell viability, cellular double-stranded DNA content, oxidative stress, steatosis, cellular ATP content, caspase-3 activity, the mitochondrial permeability transition and induction of cytochrome P450 activity. Significant concentration-dependent differences from control were observed in some endpoints at medium concentrations of 10 microg ml(-1) and above. These in vitro findings were compared with comparable endpoints obtained from an in vivo study of androstenedione toxicity in female Sprague-Dawley rats. Of the eight endpoints that could be compared between the two studies, only three (lipid accumulation, ATP depletion and P450 activity) appeared to be concordant. This suggests that, under the experimental conditions used, the clone-9 cells were not a good model for androstenedione hepatotoxicity.
Division of Toxicology, Office of Applied Research and Safety Assessment, U. S. Food and Drug Administration, Laurel, MD 20708, USA. saura.sahu@fda.hhs.gov
Saura C
SC
Sahu
Paddy L
PL
Wiesenfeld
Chung S
CS
Kim
Ivan A
IA
Ross
Philip P
PP
Sapienza
Richard
R
Newell
Michael W
MW
O'Donnell
Thomas J
TJ
Flynn
D000255:Adenosine Triphosphate / Q000378:metabolism; D000735:Androstenedione / Q000633:toxicity*; D000818:Animals; D015415:Biomarkers; D053148:Caspase 3 / Q000378:metabolism; D002460:Cell Line; D056486:Chemical and Drug Induced Liver Injury / Q000473:pathology*; D002999:Clone Cells; D004247:DNA / Q000096:biosynthesis / Q000235:genetics; D004798:Enzymes / Q000097:blood / Q000378:metabolism; D005260:Female; D022781:Hepatocytes / Q000187:drug effects / Q000201:enzymology; D008099:Liver / Q000166:cytology / Q000187:drug effects / Q000473:pathology; D008954:Models, Biological; D018384:Oxidative Stress / Q000187:drug effects; D051381:Rats; D017207:Rats, Sprague-Dawley; D015203:Reproducibility of Results
D015415:Biomarkers; D004798:Enzymes; D000735:Androstenedione; D000255:Adenosine Triphosphate; D004247:DNA; D053148:Caspase 3