Memantine Is Protective against Cytotoxicity Caused by Lead and Quinolinic Acid in Cultured Rat Embryonic Hippocampal Cells

Chem Res Toxicol. 2019 Jun 17;32(6):1134-1143. doi: 10.1021/acs.chemrestox.8b00421. Epub 2019 Apr 15.

Abstract

Quinolinic acid (QA) is an excitotoxic metabolite of the kynurenine pathway of tryptophan metabolism produced in response to inflammation and oxidative stress. Lead (Pb) causes oxidative stress and thus may produce neurotoxicity by increasing QA production. We investigated the in vitro cytotoxic effects of Pb and QA and the protective effects of the NMDA receptor antagonist memantine. Primary cultures of embryonic hippocampal cells from Wistar rats were treated with different concentrations of Pb, QA, and Pb + QA with and without memantine. Cell viability was determined by 3-(4,5-dimethylthiazol-2-yl)-2,5-diphenyltetrazolium bromide (MTT). Apoptosis was analyzed by flow cytometry after Annexin-V/propidium iodide staining. The numbers of immunostained neurons (with β3-Tubulin; Tuj1) and astrocytes (with glial fibrillary acidic protein) were counted. Pb at 20 μg/dL (0.97 μM) and QA at 500 nM concentrations showed significant cytotoxic effects, as evidenced by decreased cell viability, increased apoptosis, and a decrease in the number of both astrocytes and neurons. The combination of Pb and QA showed significant synergistic apoptotic effects at lower doses. Memantine (500 nM) was largely protective against the cytotoxic effects of both Pb and QA, suggesting that Pb's and QA's cytotoxicity involves NMDA receptor activation. Whereas the neuroprotection by memantine from QA-induced toxicity has been previously reported, this is the first study reporting the protection by memantine against Pb-induced cytotoxicity in cultured hippocampal cells. Protection by memantine against these neurotoxicants in vivo needs to be investigated.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Animals
  • Apoptosis / drug effects
  • Astrocytes / drug effects*
  • Astrocytes / pathology
  • Cell Survival / drug effects
  • Cells, Cultured
  • Dose-Response Relationship, Drug
  • Female
  • Hippocampus / cytology*
  • Hippocampus / drug effects*
  • Hippocampus / pathology
  • Lead / toxicity*
  • Male
  • Memantine / pharmacology*
  • Neurons / drug effects*
  • Neurons / pathology
  • Protective Agents / pharmacology*
  • Quinolinic Acid / toxicity*
  • Rats
  • Rats, Wistar
  • Receptors, N-Methyl-D-Aspartate / agonists
  • Structure-Activity Relationship

Substances

  • Protective Agents
  • Receptors, N-Methyl-D-Aspartate
  • Lead
  • Quinolinic Acid
  • Memantine