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Induction of Eryptosis in Red Blood Cells Using a Calcium Ionophore
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Triggering of programmed erythrocyte death by alantolactone
Kousi Alzoubi1, Salvatrice Calabrò2, Jasmin Egler3
1Department of Physiology, University of Tübingen, Gmelinstr. 5, 72076 Tuebingen, Germany. kossai.z@gmail.com.
Toxins
|December 24, 2014
Summary
Alantolactone, a compound known to induce cancer cell death, also triggers suicidal erythrocyte death, or eryptosis. This process involves cell shrinkage, phosphatidylserine exposure, increased intracellular calcium, and oxidative stress.
Area of Science:
- Biochemistry
- Cell Biology
- Toxicology
Background:
- Alantolactone exhibits anti-cancer properties by inducing apoptosis in tumor cells.
- Eryptosis, or suicidal erythrocyte death, is characterized by cell shrinkage and phosphatidylserine exposure.
- Eryptosis is triggered by increased cytosolic calcium and oxidative stress.
Purpose of the Study:
- To investigate whether alantolactone induces eryptosis in human erythrocytes.
- To determine the cellular mechanisms involved in alantolactone-induced eryptosis.
Main Methods:
- Erythrocyte volume assessed by forward scatter.
- Phosphatidylserine exposure measured by FITC-annexin-V binding.
- Intracellular calcium levels ([Ca2+]i) quantified using Fluo3 fluorescence.
- Ceramide abundance detected via fluorescent antibody binding.
- Oxidative stress evaluated using DCFDA fluorescence.
Main Results:
- Alantolactone (≥20 μM) reduced erythrocyte forward scatter and increased annexin-V binding after 48 hours.
- Alantolactone (60 μM) significantly elevated Fluo3 fluorescence and ceramide abundance.
- Alantolactone (≥40 μM) increased DCFDA fluorescence, indicating oxidative stress.
- Extracellular calcium removal did not significantly alter alantolactone's effect on annexin-V binding.
Conclusions:
- Alantolactone stimulates eryptosis in human erythrocytes.
- Alantolactone-induced eryptosis is associated with increased intracellular calcium, ceramide abundance, and oxidative stress.
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