Hepatotoxicity of piperazine designer drugs: Comparison of different in vitro models

Toxicol In Vitro. 2015 Aug;29(5):987-96. doi: 10.1016/j.tiv.2015.04.001. Epub 2015 Apr 9.

Abstract

Piperazine derived drugs emerged on the drug market in the last decade. The aim of this study was to investigate in vitro the potential hepatotoxicity of the designer drugs N-benzylpiperazine (BZP), 1-(3-trifluoromethylphenyl)piperazine (TFMPP), 1-(4-methoxyphenyl)piperazine (MeOPP) and 1-(3,4-methylenedioxybenzyl)piperazine (MDBP) in two human hepatic cell lines (HepaRG and HepG2) and in primary rat hepatocytes. Cell death was evaluated by the MTT assay, after 24 h-incubations. Among the tested drugs, TFMPP was the most cytotoxic. HepaRG cells and primary hepatocytes revealed to be the most and the least resistant cellular models, respectively. To ascertain whether the CYP450 metabolism could explain their higher susceptibility, primary hepatocytes were co-incubated with the piperazines and the CYP450 inhibitors metyrapone and quinidine, showing that CYP450-mediated metabolism contributes to the detoxification of these drugs. Additionally, the intracellular contents of reactive species, ATP, reduced (GSH) and oxidized (GSSG) glutathione, changes in mitochondrial membrane potential (Δψm) and caspase-3 activation were further evaluated in primary cells. Overall, an increase in reactive species formation, followed by intracellular GSH and ATP depletion, loss of Δψm and caspase-3 activation was observed for all piperazines, in a concentration-dependent manner. In conclusion, piperazine designer drugs produce hepatic detrimental effects that can vary in magnitude among the different analogues.

Keywords: HepG2 cells; HepaRG cells; Hepatotoxicity; Piperazine designer drugs; Primary rat hepatocytes.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Adenosine Triphosphate / metabolism
  • Animals
  • Caspase 3 / metabolism
  • Cell Line, Tumor
  • Cell Survival / drug effects
  • Cells, Cultured
  • Cytochrome P-450 Enzyme Inhibitors / pharmacology
  • Designer Drugs / toxicity*
  • Glutathione / metabolism
  • Glutathione Disulfide / metabolism
  • Hepatocytes / drug effects*
  • Hepatocytes / metabolism
  • Humans
  • Male
  • Membrane Potential, Mitochondrial / drug effects
  • Metyrapone / pharmacology
  • Piperazines / toxicity*
  • Quinidine / pharmacology
  • Rats, Wistar
  • Reactive Oxygen Species / metabolism

Substances

  • Cytochrome P-450 Enzyme Inhibitors
  • Designer Drugs
  • Piperazines
  • Reactive Oxygen Species
  • Adenosine Triphosphate
  • Caspase 3
  • Glutathione
  • Quinidine
  • Glutathione Disulfide
  • Metyrapone