Archives of Toxicology

, Volume 86, Issue 5, pp 741–752

Polyphyllin D induces apoptosis in human erythrocytes through Ca2+ rise and membrane permeabilization

  • Minghui Gao
  • K. L. Cheung
  • Irene P. Lau
  • W. S. Yu
  • K. P. Fung
  • Biao Yu
  • J. F. Loo
  • S. K. Kong
In vitro systems

DOI: 10.1007/s00204-012-0808-4

Cite this article as:
Gao, M., Cheung, K.L., Lau, I.P. et al. Arch Toxicol (2012) 86: 741. doi:10.1007/s00204-012-0808-4

Abstract

Polyphyllin D (PD) is a potent anticancer agent isolated from a traditional medicinal herb Paris polyphylla that has been used in China for many years to treat cancer. PD is not a substrate of p-glycoprotein, and it can bypass the multi-drug resistance in cancer cell line R-HepG2. However, the effect of PD on the induction of cell death in human erythrocytes remains unknown. Given that PD is a small molecule that can depolarize the mitochondrial membrane potential and release apoptosis-inducing factor (AIF) in isolated mitochondria, we hypothesized that the apoptogenic effect of PD in human erythrocytes devoid of mitochondria would be minimal. This study therefore tried to evaluate the in vitro effect of PD on hemolysis and apoptosis in human erythrocytes. Apoptosis in human red blood cells (RBCs), also known as eryptosis or erythroptosis, after PD treatment was determined by flow cytometry and confocal microscopy for the phosphatidyl-serine externalization and other apoptosis feature events. False to our prediction, PD caused hemolysis and eryptosis/erythroptosis in human RBCs. Mechanistically, elevation in the cytosolic Ca2+ ion level seems to be a key but not the only mediator in the PD-mediated eryptosis/erythroptosis because depletion of the external Ca2+ could not eliminate the PD effect. Also, PD was able to permeabilize the membrane of RBC ghosts in a way similar to digitonin. Taken together, we report here for the first time the toxicity of PD in human RBCs as well as its underlying mechanism for the hemolysis and eryptosis/erythroptosis.

Keywords

Polyphyllin D Eryptosis Erythroptosis Erythrocytes Caspase-3 Ca2+ 

Abbreviations

AIF

Apoptosis-inducing factor

[Ca2+]i

Intracellular free calcium ion concentration

FSC

Forward scatter

Hb

Hemoglobin

MDR

Multi-drug resistance

PD

Polyphyllin D

Pgp

P-glycoprotein

PS

Phosphatidyl-serine

RBCs

Red blood cells

STS

Staurosporine

Supplementary material

204_2012_808_MOESM1_ESM.pdf (569 kb)
Supplementary material 1 (PDF 569 kb)

Copyright information

© Springer-Verlag 2012

Authors and Affiliations

  • Minghui Gao
    • 1
  • K. L. Cheung
    • 1
  • Irene P. Lau
    • 1
  • W. S. Yu
    • 1
  • K. P. Fung
    • 2
  • Biao Yu
    • 3
  • J. F. Loo
    • 1
  • S. K. Kong
    • 1
  1. 1.Programme of Biochemistry, School of Life SciencesThe Chinese University of Hong KongShatin, NTChina
  2. 2.School of Biomedical SciencesThe Chinese University of Hong KongShatinHong Kong
  3. 3.State Key Laboratory of Bio-organic and Natural Products Chemistry, Shanghai Institute of Organic ChemistryChinese Academy of SciencesShanghaiChina

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