The effects of sulfhydryl blockers and metal ions on nickel accumulation by rat primary hepatocyte cultures.
Previously, we found that nickel (Ni) accumulation by rat hepatocytes involves Ca channel transport processes. However, other mechanisms responsible for Ni accumulation are still unclear. Therefore, in the present study we examined the effects of sulfhydryl (SH) blockers on Ni accumulation by hepatocytes. Hepatocytes were exposed to various concentrations of N-ethylmaleimide (NEM) (0.5, 1 or 2 mM) or monoiodoacetic acid (MIA) (0.1, 0.25 or 0.5 mM), potent blockers of SH ligands, for 30 min and subsequently exposed to 10 microM NiCl(2) for an additional 60 min. Pretreatment with NEM and MIA enhanced the Ni accumulation by hepatocytes to maximum of 156 and 73%, respectively. The effects of essential and nonessential metal ions on Ni accumulation were also investigated. Pretreatment with 10 microM of Cu, Zn, Co, Cd and Mn, decreased Ni accumulation by 46, 30, 20, 18 and 11%, respectively. In contrast, pretreatment with Hg (10 and 20 microM) enhanced the Ni accumulation by almost 81 and 140%, respectively. Furthermore, significant decrease in SH concentration in the hepatocyte membrane was observed by the treatment with NEM, MIA and Hg, but not with Cu, Zn and Cd. These results suggest that Ni accumulation by hepatocytes does not appear to be dependent on the SH carrier-mediated transport processes, and that to block the SH ligands in the plasma membrane may facilitate the Ni crossing of the cell membrane.
Shimada
H
H
Department of Hygienic Chemistry, Faculty of Pharmaceutical Sciences, Kumamoto University, 5-1 Oe-honmachi, Kumamoto 862-0973, Japan. hshimada@gpo.kumamoto-u.ac.jp
Funakoshi
T
T
Inoue
T
T
Kojima
S
S
D000818:Animals; D002413:Cations, Divalent; D002462:Cell Membrane / Q000187:drug effects / Q000378:metabolism; D004305:Dose-Response Relationship, Drug; D004347:Drug Interactions; D005033:Ethylmaleimide / Q000494:pharmacology*; D022781:Hepatocytes / Q000187:drug effects* / Q000378:metabolism; D019807:Iodoacetic Acid / Q000494:pharmacology*; D008297:Male; D008670:Metals / Q000494:pharmacology*; D009532:Nickel / Q000493:pharmacokinetics*; D051381:Rats; D017208:Rats, Wistar; D013439:Sulfhydryl Reagents / Q000494:pharmacology*
D002413:Cations, Divalent; D008670:Metals; D013439:Sulfhydryl Reagents; C022838:nickel chloride; D009532:Nickel; D005033:Ethylmaleimide; D019807:Iodoacetic Acid