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Ceranib-2-induced suicidal erythrocyte death
Elena Signoretto1,2, Jens Zierle1, Abdulla Al Mamun Bhuyan1
1Departments of Physiology and Cardiology & Cardiovascular Medicine, Eberhard-Karls-University of Tuebingen, Tuebingen, Germany.
Cell Biochemistry and Function
|June 14, 2016
Summary
The ceramidase inhibitor Ceranib-2 triggers eryptosis, a form of programmed cell death in erythrocytes. This occurs due to increased ceramide levels and oxidative stress, leading to cell membrane damage.
Area of Science:
- Cell Biology
- Hematology
- Biochemistry
Background:
- Ceramide induces apoptosis in nucleated cells and eryptosis in erythrocytes.
- Eryptosis involves cell shrinkage, membrane scrambling, and phosphatidylserine translocation.
- Increased cytosolic calcium and oxidative stress are known eryptosis stimulators.
Purpose of the Study:
- To investigate whether the ceramidase inhibitor Ceranib-2 induces eryptosis in human erythrocytes.
- To elucidate the mechanisms underlying Ceranib-2-induced eryptosis.
Main Methods:
- Flow cytometry was used to assess phosphatidylserine exposure (Annexin-V binding), cell volume (forward scatter), cytosolic calcium ([Ca(2+)]i), and reactive oxygen species (ROS).
- Ceramide abundance was quantified using specific antibodies.
- Hemolysis was measured by hemoglobin concentration in the supernatant.
Main Results:
- Ceranib-2 exposure (48 hours) significantly increased Annexin-V binding and hemolysis in a dose-dependent manner.
- Ceranib-2 elevated cytosolic calcium, ROS levels, and ceramide abundance.
- The eryptosis-inducing effect of Ceranib-2 was independent of extracellular calcium influx.
Conclusions:
- Ceranib-2 induces eryptosis by triggering phospholipid scrambling of the erythrocyte membrane.
- Increased ceramide abundance and oxidative stress are key mechanisms in Ceranib-2-induced eryptosis.
- Calcium entry is not required for Ceranib-2-mediated eryptosis.