The
sesquiterpene alantolactone counteracts
malignancy, an effect at least in part due to stimulation of suicidal death or apoptosis of
tumor cells. Signaling of
alantolactone induced apoptosis involves altered gene expression and mitochondrial depolarization. Erythrocytes lack mitochondria and nuclei but may enter suicidal death or eryptosis, which is characterized by cell shrinkage and cell membrane scrambling with
phosphatidylserine exposure at the erythrocyte surface. Cellular mechanisms involved in triggering of eryptosis include increase of cytosolic Ca2+-activity ([Ca2+]i) and oxidative stress. The present study explored, whether
alantolactone stimulates eryptosis. To this end, erythrocyte volume was estimated from forward scatter,
phosphatidylserine-exposure at the erythrocyte surface from
FITC-
annexin-V-binding, [Ca2+]i from Fluo3-fluorescence,
ceramide abundance from binding of fluorescent
antibodies, and oxidative stress from 2',7'-dichlorodihydrofluorescein-diacetate (
DCFDA) fluorescence. As a result, a 48 h exposure of human erythrocytes to
alantolactone (≥20 μM) significantly decreased erythrocyte forward scatter and increased the percentage of
annexin-V-binding cells.
Alantolactone significantly increased Fluo3 fluorescence (60 μM),
ceramide abundance (60 μM) and
DCFDA fluorescence (≥40 μM). The effect of
alantolactone (60 μM) on
annexin-V-binding was not significantly modified by removal of extracellular Ca2+. In conclusion,
alantolactone stimulates suicidal erythrocyte death or eryptosis, an effect paralleled by increase of [Ca2+]i,
ceramide abundance and oxidative stress.