High-concentration homocysteine inhibits mitochondrial respiration function and production of reactive oxygen species in neuron cells

J Stroke Cerebrovasc Dis. 2020 Oct;29(10):105109. doi: 10.1016/j.jstrokecerebrovasdis.2020.105109. Epub 2020 Jul 28.


Objective: Homocysteine plays critical roles in cellular redox homeostasis, and hyperhomocysteinemia has been associated with multiple diseases, including neurological disorders involving reactive oxygen species-inducing and pro-inflammatory effects of homocysteine that are related to mitochondria. This study investigated the role of homocysteine in regulating mitochondria of neuron cell lines.

Methods: Neuron cells were pre-treated with homocysteine, and then flow cytometry was used to detect reactive oxygen species production and mitochondrial membrane potential, while Seahorse XFp Mito stress assay was used to comprehensively analyze mitochondrial function.

Results: The experimental results showed that high-concentration homocysteine diminished carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone-stimulated oxygen consumption rate and mitochondrial spare respiration capacity in a time- and concentration-dependent manner, and homocysteine also reduced reactive oxygen species in cultured neuron cell lines while no changes in mitochondrial membrane potential were observed.

Conclusion: These results indicate that homocysteine diminished mitochondrial respiration function in neuron cell lines mediated by its reactive oxygen species-reducing effects, which may underlie the association between hyperhomocysteinemia and human diseases.

Keywords: Carbonyl cyanide-4 (trifluoromethoxy) phenylhydrazone (FCCP); Homocysteine (Hcy); Mitochondrial respiration function; Neuron cell lines; Oxygen consumption rate (OCR); reactive oxygen species (ROS).

MeSH terms

  • Animals
  • Cell Line
  • Cell Respiration / drug effects
  • Dose-Response Relationship, Drug
  • Homocysteine / toxicity*
  • Mice
  • Mitochondria / drug effects*
  • Mitochondria / metabolism
  • Mitochondria / pathology
  • Neurons / drug effects*
  • Neurons / metabolism
  • Neurons / pathology
  • Rats
  • Reactive Oxygen Species / metabolism*
  • Time Factors


  • Reactive Oxygen Species
  • Homocysteine