Diethanolamine induces hepatic choline deficiency in mice.
The purpose of the present experiments was to test the hypothesis that diethanolamine (DEA), an alkanolamine shown to be hepatocarcinogenic in mice, induces hepatic choline deficiency and to determine whether altered choline homeostasis was causally related to the carcinogenic outcome. To examine this hypothesis, the biochemical and histopathological changes in male B6C3F1 mice made choline deficient by dietary deprivation were first determined. Phosphocholine (PCho), the intracellular storage form of choline was severely depleted, decreasing to about 20% of control values with 2 weeks of dietary choline deficiency. Other metabolites, including choline, glycerophosphocholine (GPC), and phosphatidylcholine (PC) also decreased. Hepatic concentrations of S-adenosylmethionine (SAM) decreased, whereas levels of S-adenosylhomocysteine (SAH) increased. Despite these biochemical changes, fatty liver, which is often associated with choline deficiency, was not observed in the mice. The dose response, reversibility, and strain-dependence of the effects of DEA on choline metabolites were studied. B6C3F1 mice were dosed dermally with DEA (0, 10, 20, 40, 80, and 160 mg/kg) for 4 weeks (5 days/week). Control animals received either no treatment or dermal application of 95% ethanol (1.8 ml/kg). PCho was most sensitive to DEA treatment, decreasing at dosages of 20 mg/kg and higher and reaching a maximum 50% depletion at 160 mg/kg/day. GPC, choline, and PC also decreased in a dose-dependent manner. At 80 and 160 mg/kg/day, SAM levels decreased while SAH levels increased in liver. A no-observed effect level (NOEL) for DEA-induced changes in choline homeostasis was 10 mg/kg/day. Choline metabolites, SAM and SAH returned to control levels in mice dosed at 160 mg/kg for 4 weeks and allowed a 2-week recovery period prior to necropsy. In a manner similar to dietary choline deficiency, no fatty change was observed in the liver of DEA-treated mice. In C57BL/6 mice, DEA treatment (160 mg/kg) also decreased PCho concentrations, without affecting hepatic SAM levels, suggesting that strain-specific differences in intracellular methyl group regulation may influence carcinogenic outcome with DEA treatment. Finally, in addition to the direct effects of DEA on choline homeostasis, dermal application of 95% ethanol for 4 weeks decreased hepatic betaine levels, suggesting that the use of ethanol as a vehicle for dermal application of DEA may exacerbate or confound the biochemical actions of DEA alone. Collectively, the results demonstrate that DEA treatment causes a spectrum of biochemical changes consistent with choline deficiency in mice and demonstrate a clear dose concordance between DEA-induced choline deficiency and hepatocarcinogenic outcome.
Lehman-McKeeman
Lois D
LD
Central Product Safety Division, Miami Valley Laboratories, Procter and Gamble Co., Cincinnati, Ohio 45253, USA. lois.lehman-mckeeman@bms.com
Gamsky
Elizabeth A
EA
Hicks
Sarah M
SM
Vassallo
Jeffrey D
JD
Mar
Mei-Heng
MH
Zeisel
Steven H
SH
D000279:Administration, Cutaneous; D000818:Animals; D001622:Betaine / Q000378:metabolism; D002273:Carcinogens / Q000008:administration & dosage / Q000633:toxicity*; D002796:Choline Deficiency / Q000139:chemically induced* / Q000378:metabolism / Q000473:pathology; D004305:Dose-Response Relationship, Drug; D004357:Drug Synergism; D000431:Ethanol / Q000633:toxicity; D004983:Ethanolamines / Q000008:administration & dosage / Q000633:toxicity*; D005997:Glycerylphosphorylcholine / Q000378:metabolism; D008099:Liver / Q000187:drug effects* / Q000378:metabolism / Q000473:pathology; D008297:Male; D051379:Mice; D008810:Mice, Inbred C57BL; D019055:No-Observed-Adverse-Effect Level; D010713:Phosphatidylcholines / Q000378:metabolism; D010767:Phosphorylcholine / Q000378:metabolism; D012435:S-Adenosylhomocysteine / Q000378:metabolism; D012436:S-Adenosylmethionine / Q000378:metabolism; D013045:Species Specificity
D002273:Carcinogens; D004983:Ethanolamines; D010713:Phosphatidylcholines; D010767:Phosphorylcholine; D000431:Ethanol; D001622:Betaine; D005997:Glycerylphosphorylcholine; D012436:S-Adenosylmethionine; D012435:S-Adenosylhomocysteine; C020283:diethanolamine