Impact of Repeated Nicotine and Alcohol Coexposure on in Vitro and in Vivo Chlorpyrifos Dosimetry and Cholinesterase Inhibition

Abstract

Chlorpyrifos (CPF) is an organophosphorus insecticide, and neurotoxicity results from inhibition of acetylcholinesterase (AChE) by its metabolite, chlorpyrifos-oxon. Routine consumption of alcohol and tobacco modifies metabolic and physiological processes impacting the metabolism and pharmacokinetics of other xenobiotics, including pesticides. This study evaluated the influence of repeated ethanol and nicotine coexposure on in vivo CPF dosimetry and cholinesterase (ChE) response (ChE- includes AChE and/or butyrylcholinesterase (BuChE)). Hepatic microsomes were prepared from groups of naive, ethanol-only (1 g/kg/d, 7 d, po), and ethanol + nicotine (1 mg/kg/d 7 d, sc)-treated rats, and the in vitro metabolism of CPF was evaluated. For in vivo studies, rats were treated with saline or ethanol (1 g/kg/d, po) + nicotine (1 mg/kg/d, sc) in addition to CPF (1 or 5 mg/kg/d, po) for 7 d. The major CPF metabolite, 3,5,6-trichloro-2-pyridinol (TCPy), in blood and urine and the plasma ChE and brain acetylcholinesterase (AChE) activities were measured in rats. There were differences in pharmacokinetics, with higher TCPy peak concentrations and increased blood TCPy AUC in ethanol + nicotine groups compared to CPF only (approximately 1.8- and 3.8-fold at 1 and 5 mg CPF doses, respectively). Brain AChE activities after ethanol + nicotine treatments showed significantly less inhibition following repeated 5 mg CPF/kg dosing compared to CPF only (96 ± 13 and 66 ± 7% of naive at 4 h post last CPF dosing, respectively). Although brain AChE activity was minimal inhibited for the 1-mg CPF/kg/d groups, the ethanol + nicotine pretreatment resulted in a similar trend (i.e., slightly less inhibition). No marked differences were observed in plasma ChE activities due to the alcohol + nicotine treatments. In vitro, CPF metabolism was not markedly affected by repeated ethanol or both ethanol + nicotine exposures. Compared with a previous study of nicotine and CPF exposure, there were no apparent additional exacerbating effects due to ethanol coexposure.

Acknowledgments

Although Drs. Timchalk and Poet have received funding from the Dow Chemical Company, the manufacturer of chlorpyrifos, to conduct research, the Dow Chemical Company did not have any involvement in the study design, analysis, and interpretation of data, in the writing of the report, or in the decision to submit this paper for publication. We gratefully acknowledge Dr. J. Campbell for valuable discussions. Although the research described in this article has been funded by the Centers for Disease Control and Prevention (CDC)/National Institute for Occupational Safety and Health (NIOSH) [CDC/NIOSH grant (R01-OH003629) to Dr. Timchalk], it has not been subject to any CDC/NIOSH review and therefore does not necessarily reflect the official views of the agencies; its contents are solely the responsibility of the authors and no public endorsement should be inferred.