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Updated: Dec 12, 2025
Measurement of Heme Synthesis Levels in Mammalian Cells
Published on: July 9, 2015
Homocysteine-induced decrease in HUVEC cells' resistance to oxidative stress is mediated by Akt-dependent changes in
Andzelika Borkowska1, Wieslaw Ziolkowski2, Katarzyna Kaczor1
1Department of Bioenergetics and Physiology of Exercise, Medical University of Gdansk, 1 Debinki St., 80-210, Gdańsk, Poland.
Insights
Homocysteine (Hcy) increases cellular ferritin levels in HUVEC cells by affecting the Akt-FOXO3a pathway, impacting iron metabolism and increasing oxidative stress. This finding is crucial for understanding cardiovascular and neurodegenerative diseases.
Area of Science:
- Biochemistry
- Cell Biology
- Toxicology
Background:
- Hyperhomocysteinemia is a risk factor for cardiovascular and neurodegenerative diseases.
- Iron may mediate homocysteine (Hcy) toxicity.
Purpose of the Study:
- Investigate the effect of Hcy on iron metabolism in HUVEC and SH-SY5Y cells.
- Elucidate the molecular mechanisms underlying Hcy-induced changes in iron metabolism.
Main Methods:
- HUVEC and SH-SY5Y cells were treated with 3 mM Hcy.
- Analyzed ferritin expression, Akt kinase activity, and FOXO3a signaling.
- Utilized siRNA to investigate the roles of Akt and FOXO3a.
Main Results:
- Hcy upregulated ferritins L and H in HUVEC cells in a time-dependent manner.
- Hcy decreased active Akt kinase in HUVEC cells, preceding ferritin changes.
- Akt activation increased ferritin levels, while FOXO3a inhibition decreased them.
- Hcy-treated HUVEC cells showed increased hydrogen peroxide cytotoxicity.
Conclusions:
- Hcy increases cellular ferritin levels via the Akt-FOXO3a signaling pathway.
- This pathway modulation affects iron metabolism and cellular response to oxidative stress.
Purpose:
Hyperhomocysteinemia is an independent risk factor for cardiovascular diseases and also promotes neuronal death in various neurodegenerative diseases. There is evidence that iron can mediate homocysteine (Hcy) toxicity. Thus, the aim of this study was to investigate the effect of Hcy on iron metabolism in HUVEC and SH-SY5Y cells.
Methods:
HUVEC and SH-SY5Y cells were treated with 3 mM Hcy for a defined time.
Results:
We demonstrate that Hcy induced the upregulation of ferritins type L and H in HUVEC cells in a time-dependent manner and had no effect on the ferritins in SH-SY5Y cells. The change in ferritin expression was preceded by a significant decrease in the cellular level of the active form of Akt kinase in HUVEC but not in SH-SY5Y cells. An increase in ferritin L and H protein levels was observed in the Akt1, Akt2, Akt3 siRNA transfected cells, while in the cells transfected with FOXO3a siRNA, a decrease in both ferritins levels was noticed. Moreover, in the HUVEC cells treated with Hcy for 6 days, the active form of kinase Akt returned to the control level and it was accompanied by a drop in ferritin L and H protein levels. Cytotoxicity of hydrogen peroxide significantly increased in HUVEC cells pre-treated with Hcy for 24 h.
Conclusions:
These data indicate that Hcy induces an increase in cellular ferritin level, and the process is mediated by alterations in Akt-FOXO3a signaling pathway.
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