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Piperlongumine-induced phosphatidylserine translocation in the erythrocyte membrane.

Bissinger R, Malik A, Warsi J, Jilani K, Lang F - Toxins (Basel) (2014)

Bottom Line: Piperlongumine did not significantly modify [Ca²⁺]i and the effect was not dependent on presence of extracellular Ca²⁺.Piperlongumine significantly increased ROS formation and ceramide abundance.Piperlongumine triggers cell membrane scrambling, an effect independent from entry of extracellular Ca²⁺ but at least partially due to ROS and ceramide formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Eberhard-Karls-University of Tuebingen, Gmelinstr. 5, 72076 Tuebingen, Germany. ro.bissinger@gmx.de.

ABSTRACT

Background: Piperlongumine, a component of Piper longum fruit, is considered as a treatment for malignancy. It is effective by inducing apoptosis. Mechanisms involved in the apoptotic action of piperlongumine include oxidative stress and activation of p38 kinase. In analogy to apoptosis of nucleated cells, erythrocytes may undergo eryptosis, the suicidal death of erythrocytes characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine-exposure at the erythrocyte surface. Signaling involved in eryptosis include increase of cytosolic Ca²⁺-activity ([Ca²⁺]i), formation of ceramide, oxidative stress and activation of p38 kinase.

Methods: Cell volume was estimated from forward scatter, phosphatidylserine-exposure from annexin V binding, [Ca²⁺]i from Fluo3 fluorescence, reactive oxygen species from 2',7'-dichlorodihydrofluorescein-diacetate fluorescence, and ceramide abundance from binding of fluorescent antibodies in flow cytometry.

Results: A 48 h exposure to piperlongumine (30 µM) was followed by significant decrease of forward scatter and increase of annexin-V-binding. Piperlongumine did not significantly modify [Ca²⁺]i and the effect was not dependent on presence of extracellular Ca²⁺. Piperlongumine significantly increased ROS formation and ceramide abundance.

Conclusions: Piperlongumine triggers cell membrane scrambling, an effect independent from entry of extracellular Ca²⁺ but at least partially due to ROS and ceramide formation.

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Related in: MedlinePlus

Effect of piperlongumine on phosphatidylserine exposure (A) Original histogram of annexin V binding of erythrocytes following exposure for 48 h to Ringer solution without (grey shadow) and with (black line) presence of 30 µM piperlongumine; (B) Arithmetic means ± SEM (n = 15) of erythrocyte annexin-V-binding following incubation for 48 h to Ringer solution without (white bar) or with (black bars) presence of piperlongumine (5–30 µM) or, for comparison, DMSO (0.1%) alone. * (p < 0.05), *** (p < 0.001) indicate significant differences from the absence of piperlongumine (ANOVA).
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toxins-06-02975-f001: Effect of piperlongumine on phosphatidylserine exposure (A) Original histogram of annexin V binding of erythrocytes following exposure for 48 h to Ringer solution without (grey shadow) and with (black line) presence of 30 µM piperlongumine; (B) Arithmetic means ± SEM (n = 15) of erythrocyte annexin-V-binding following incubation for 48 h to Ringer solution without (white bar) or with (black bars) presence of piperlongumine (5–30 µM) or, for comparison, DMSO (0.1%) alone. * (p < 0.05), *** (p < 0.001) indicate significant differences from the absence of piperlongumine (ANOVA).

Mentions: The present study explored the putative effect of piperlongumine on eryptosis, the suicidal erythrocyte death. Hallmarks of eryptosis include breakdown of phosphatidylserine asymmetry of the erythrocyte cell membrane with subsequent translocation of phosphatidylserine to the cell surface. Phosphatidylserine exposing erythrocytes were identified by annexin-V-binding in flow cytometry. As illustrated in Figure 1, a 48 h exposure to piperlongumine increased the percentage of annexin-V-binding erythrocytes, an effect reaching statistical significance at 15 µM piperlongumine concentration.


Piperlongumine-induced phosphatidylserine translocation in the erythrocyte membrane.

Bissinger R, Malik A, Warsi J, Jilani K, Lang F - Toxins (Basel) (2014)

Effect of piperlongumine on phosphatidylserine exposure (A) Original histogram of annexin V binding of erythrocytes following exposure for 48 h to Ringer solution without (grey shadow) and with (black line) presence of 30 µM piperlongumine; (B) Arithmetic means ± SEM (n = 15) of erythrocyte annexin-V-binding following incubation for 48 h to Ringer solution without (white bar) or with (black bars) presence of piperlongumine (5–30 µM) or, for comparison, DMSO (0.1%) alone. * (p < 0.05), *** (p < 0.001) indicate significant differences from the absence of piperlongumine (ANOVA).
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4210880&req=5

toxins-06-02975-f001: Effect of piperlongumine on phosphatidylserine exposure (A) Original histogram of annexin V binding of erythrocytes following exposure for 48 h to Ringer solution without (grey shadow) and with (black line) presence of 30 µM piperlongumine; (B) Arithmetic means ± SEM (n = 15) of erythrocyte annexin-V-binding following incubation for 48 h to Ringer solution without (white bar) or with (black bars) presence of piperlongumine (5–30 µM) or, for comparison, DMSO (0.1%) alone. * (p < 0.05), *** (p < 0.001) indicate significant differences from the absence of piperlongumine (ANOVA).
Mentions: The present study explored the putative effect of piperlongumine on eryptosis, the suicidal erythrocyte death. Hallmarks of eryptosis include breakdown of phosphatidylserine asymmetry of the erythrocyte cell membrane with subsequent translocation of phosphatidylserine to the cell surface. Phosphatidylserine exposing erythrocytes were identified by annexin-V-binding in flow cytometry. As illustrated in Figure 1, a 48 h exposure to piperlongumine increased the percentage of annexin-V-binding erythrocytes, an effect reaching statistical significance at 15 µM piperlongumine concentration.

Bottom Line: Piperlongumine did not significantly modify [Ca²⁺]i and the effect was not dependent on presence of extracellular Ca²⁺.Piperlongumine significantly increased ROS formation and ceramide abundance.Piperlongumine triggers cell membrane scrambling, an effect independent from entry of extracellular Ca²⁺ but at least partially due to ROS and ceramide formation.

View Article: PubMed Central - PubMed

Affiliation: Department of Physiology, Eberhard-Karls-University of Tuebingen, Gmelinstr. 5, 72076 Tuebingen, Germany. ro.bissinger@gmx.de.

ABSTRACT

Background: Piperlongumine, a component of Piper longum fruit, is considered as a treatment for malignancy. It is effective by inducing apoptosis. Mechanisms involved in the apoptotic action of piperlongumine include oxidative stress and activation of p38 kinase. In analogy to apoptosis of nucleated cells, erythrocytes may undergo eryptosis, the suicidal death of erythrocytes characterized by cell shrinkage and cell membrane scrambling with phosphatidylserine-exposure at the erythrocyte surface. Signaling involved in eryptosis include increase of cytosolic Ca²⁺-activity ([Ca²⁺]i), formation of ceramide, oxidative stress and activation of p38 kinase.

Methods: Cell volume was estimated from forward scatter, phosphatidylserine-exposure from annexin V binding, [Ca²⁺]i from Fluo3 fluorescence, reactive oxygen species from 2',7'-dichlorodihydrofluorescein-diacetate fluorescence, and ceramide abundance from binding of fluorescent antibodies in flow cytometry.

Results: A 48 h exposure to piperlongumine (30 µM) was followed by significant decrease of forward scatter and increase of annexin-V-binding. Piperlongumine did not significantly modify [Ca²⁺]i and the effect was not dependent on presence of extracellular Ca²⁺. Piperlongumine significantly increased ROS formation and ceramide abundance.

Conclusions: Piperlongumine triggers cell membrane scrambling, an effect independent from entry of extracellular Ca²⁺ but at least partially due to ROS and ceramide formation.

Show MeSH
Related in: MedlinePlus