Tumor necrosis factor-α induces caspase-independent cell death in human neutrophils via reactive oxidants and associated with calpain activity

Hao Cheng Chen, Chih Jan Wang, Chun Liang Chou, Shu Min Lin, Chien Da Huang, Ting Yu Lin, Chun Hua Wang, Horng Chyuan Lin, Chih Teng Yu, Han Pin Kuo, Chien Ying Liu

Research output: Contribution to journalArticle

10 Citations (Scopus)

Abstract

Apoptosis mediated by caspase activation is important in the neutrophil homeostasis and resolution of tissue inflammation. Paradoxically, our previous study demonstrated that broad-spectrum caspase inhibition augmented tumor necrosis factor (TNF)-α-induced cell death in the human neutrophils. Therefore, we further explored the mechanisms related to the caspase-independent cell death in the neutrophils. The cell apoptosis/necrosis was determined by annexin V and propidium iodide dual staining in flow cytometry. Their morphological changes were observed under light microscopy. Fluorogenic substrates were used to measure the intracellular oxidative reactions and the activities of proteinases, calpains. Calpain inhibitors and antioxidants were used to elucidate the relationship of calpains and oxidants with the neutrophil cell death. Our results verified the caspase-independent cell death pathway in the zVAD-sensitized, TNF-α-stimulated neutrophils. Furthermore, the cell death was accompanied with increased calpain and oxidative activities in the cells. Calpain inhibitors, zLLY, as well as anti-oxidants, catalase and DMSO, were able to attenuate the cell death in the zVAD-sensitized, TNF-α-induced neutrophils. Pretreating the neutrophils with G-CSF or GM-CSF was not able to reduce the cell death. These results demonstrate that, in human neutrophils, TNF-α-induces a caspase-independent cell death signal, which is related to calpain and oxidative activities and cannot be rescued by the growth factor-related signaling mechanism.

Original languageEnglish
Pages (from-to)261-273
Number of pages13
JournalJournal of Biomedical Science
Volume13
Issue number2
DOIs
Publication statusPublished - Mar 1 2006
Externally publishedYes

Fingerprint

Calpain
Cell death
Caspases
Oxidants
Neutrophils
Cell Death
Tumor Necrosis Factor-alpha
Granulocyte-Macrophage Colony-Stimulating Factor
Apoptosis
Flow cytometry
Propidium
Annexin A5
Granulocyte Colony-Stimulating Factor
Dimethyl Sulfoxide
Fluorescent Dyes
Catalase
Optical microscopy
Microscopy
Intercellular Signaling Peptides and Proteins
Flow Cytometry

Keywords

  • Apoptosis
  • Calpain
  • Caspase
  • Neutrophil
  • Oxidant
  • Tumor necrosis factor

ASJC Scopus subject areas

  • Endocrinology, Diabetes and Metabolism
  • Molecular Biology
  • Clinical Biochemistry
  • Cell Biology
  • Biochemistry, medical
  • Pharmacology (medical)

Cite this

Tumor necrosis factor-α induces caspase-independent cell death in human neutrophils via reactive oxidants and associated with calpain activity. / Chen, Hao Cheng; Wang, Chih Jan; Chou, Chun Liang; Lin, Shu Min; Huang, Chien Da; Lin, Ting Yu; Wang, Chun Hua; Lin, Horng Chyuan; Yu, Chih Teng; Kuo, Han Pin; Liu, Chien Ying.

In: Journal of Biomedical Science, Vol. 13, No. 2, 01.03.2006, p. 261-273.

Research output: Contribution to journalArticle

Chen, Hao Cheng ; Wang, Chih Jan ; Chou, Chun Liang ; Lin, Shu Min ; Huang, Chien Da ; Lin, Ting Yu ; Wang, Chun Hua ; Lin, Horng Chyuan ; Yu, Chih Teng ; Kuo, Han Pin ; Liu, Chien Ying. / Tumor necrosis factor-α induces caspase-independent cell death in human neutrophils via reactive oxidants and associated with calpain activity. In: Journal of Biomedical Science. 2006 ; Vol. 13, No. 2. pp. 261-273.
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AU - Lin, Shu Min

AU - Huang, Chien Da

AU - Lin, Ting Yu

AU - Wang, Chun Hua

AU - Lin, Horng Chyuan

AU - Yu, Chih Teng

AU - Kuo, Han Pin

AU - Liu, Chien Ying

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N2 - Apoptosis mediated by caspase activation is important in the neutrophil homeostasis and resolution of tissue inflammation. Paradoxically, our previous study demonstrated that broad-spectrum caspase inhibition augmented tumor necrosis factor (TNF)-α-induced cell death in the human neutrophils. Therefore, we further explored the mechanisms related to the caspase-independent cell death in the neutrophils. The cell apoptosis/necrosis was determined by annexin V and propidium iodide dual staining in flow cytometry. Their morphological changes were observed under light microscopy. Fluorogenic substrates were used to measure the intracellular oxidative reactions and the activities of proteinases, calpains. Calpain inhibitors and antioxidants were used to elucidate the relationship of calpains and oxidants with the neutrophil cell death. Our results verified the caspase-independent cell death pathway in the zVAD-sensitized, TNF-α-stimulated neutrophils. Furthermore, the cell death was accompanied with increased calpain and oxidative activities in the cells. Calpain inhibitors, zLLY, as well as anti-oxidants, catalase and DMSO, were able to attenuate the cell death in the zVAD-sensitized, TNF-α-induced neutrophils. Pretreating the neutrophils with G-CSF or GM-CSF was not able to reduce the cell death. These results demonstrate that, in human neutrophils, TNF-α-induces a caspase-independent cell death signal, which is related to calpain and oxidative activities and cannot be rescued by the growth factor-related signaling mechanism.

AB - Apoptosis mediated by caspase activation is important in the neutrophil homeostasis and resolution of tissue inflammation. Paradoxically, our previous study demonstrated that broad-spectrum caspase inhibition augmented tumor necrosis factor (TNF)-α-induced cell death in the human neutrophils. Therefore, we further explored the mechanisms related to the caspase-independent cell death in the neutrophils. The cell apoptosis/necrosis was determined by annexin V and propidium iodide dual staining in flow cytometry. Their morphological changes were observed under light microscopy. Fluorogenic substrates were used to measure the intracellular oxidative reactions and the activities of proteinases, calpains. Calpain inhibitors and antioxidants were used to elucidate the relationship of calpains and oxidants with the neutrophil cell death. Our results verified the caspase-independent cell death pathway in the zVAD-sensitized, TNF-α-stimulated neutrophils. Furthermore, the cell death was accompanied with increased calpain and oxidative activities in the cells. Calpain inhibitors, zLLY, as well as anti-oxidants, catalase and DMSO, were able to attenuate the cell death in the zVAD-sensitized, TNF-α-induced neutrophils. Pretreating the neutrophils with G-CSF or GM-CSF was not able to reduce the cell death. These results demonstrate that, in human neutrophils, TNF-α-induces a caspase-independent cell death signal, which is related to calpain and oxidative activities and cannot be rescued by the growth factor-related signaling mechanism.

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