Effect of Long-term In Vitro Lithium Exposure on mRNA Levels of Claudin-3, CYP1A1, ABCG2 and GSTM3 Genes in the hCMEC/D3 Human Brain Endothelial Cell Line

Eur J Drug Metab Pharmacokinet. 2017 Dec;42(6):1013-1017. doi: 10.1007/s13318-017-0412-3.

Abstract

Background and objectives: Lithium chloride (LiCl) has been shown to improve the tightness of brain endothelial cell monolayers in vitro by inhibition of the GSK-3β enzyme, activation of the Wnt/beta-catenin pathway and regulation of tight junction (TJ) protein expression. However, the effect of LiCl on the drug transporters and drug-metabolizing enzymes has not been addressed so far. The hCMEC/D3 cell line is a validated in vitro BBB model expressing transporters and drug-metabolizing enzymes (phase 1 and 2). The present study was conducted to compare the mRNAs levels corresponding to several BBB endothelial markers in hCMEC/D3 cells with and without incubation in LiCl.

Methods: We used quantitative real-time polymerase chain reaction (qRT-PCR) to quantify the mRNA expression of 5 tight junction (TJ) proteins, 4 adhesion proteins, 5 solute carriers, 7 ATP-binding cassette (ABC) transporters, 8 cytochrome P450 (CYP) and 17 phase 2 conjugation enzymes in hCMEC/D3 cells with and without incubation in LiCl.

Results: Our study showed that LiCl treatment for 6 days at a concentration of 10 mM induced the TJ protein claudin-3, the ABC transporter BCRP/ABCG2, the cytochrome P-450 CYP1A1 and the glutathione-S-transferase GSTM3, while the other selected markers were not significantly affected.

Conclusions: Our findings provide new insights into the effects of lithium on some drug transporters and drug-metabolizing enzymes in the BBB that may have consequences in pharmacotherapy.

MeSH terms

  • ATP Binding Cassette Transporter, Subfamily G, Member 2 / genetics*
  • Biomarkers / metabolism
  • Brain / cytology*
  • Cell Line
  • Claudin-3 / genetics*
  • Cytochrome P-450 CYP1A1 / genetics*
  • Endothelial Cells / metabolism*
  • Gene Expression / drug effects
  • Glutathione Transferase / genetics*
  • Humans
  • Lithium Chloride / administration & dosage
  • Lithium Chloride / pharmacology*
  • Neoplasm Proteins / genetics*
  • RNA, Messenger / genetics
  • RNA, Messenger / metabolism*

Substances

  • ABCG2 protein, human
  • ATP Binding Cassette Transporter, Subfamily G, Member 2
  • Biomarkers
  • CLDN3 protein, human
  • Claudin-3
  • Neoplasm Proteins
  • RNA, Messenger
  • CYP1A1 protein, human
  • Cytochrome P-450 CYP1A1
  • GSTM3 protein, human
  • Glutathione Transferase
  • Lithium Chloride