The pro-inflammatory oxidant hypochlorous acid induces Bax-dependent mitochondrial permeabilisation and cell death through AIF-/EndoG-dependent pathways.

Abstract

At sites of chronic inflammation, such as in the inflamed rheumatoid joint, activated neutrophils release hydrogen peroxide (H(2)O(2)) and the enzyme myeloperoxidase to catalyse the formation of hypochlorous acid (HOCl). 3-chlorotyrosine, a marker of HOCl in vivo, has been observed in synovial fluid proteins from rheumatoid arthritis patients. However the mechanisms of HOCl-induced cytotxicity are unknown. We determined the molecular mechanisms by which HOCl induced cell death in human mesenchymal progenitor cells (MPCs) differentiated into a chondrocytic phenotype as a model of human cartilage cells and show that HOCl induced rapid Bax conformational change, mitochondrial permeability and release of intra-mitochondrial pro-apoptotic proteins which resulted in nuclear translocation of AIF and EndoG. siRNA-mediated knockdown of Bax substantially prevented mitochondrial permeability, release of intra-mitochondrial pro-apoptotic proteins. Cell death was inhibited by siRNA-mediated knockdown of Bax, AIF or EndoG. Although we observed several biochemical markers of apoptosis, caspase activation was not detected either by western blotting, fluorescence activity assays or by using caspase inhibitors to inhibit cell death. This was further supported by findings that (1) in vitro exposure of recombinant human caspases to HOCl caused significant inhibition of caspase activity and (2) the addition of HOCl to staurosporine-treated MPCs inhibited the activity of cellular caspases. Our results show for the first time that HOCl induced Bax-dependent mitochondrial permeability which led to cell death without caspase activity by processes involving AIF/EndoG-dependent pathways. Our study provides a novel insight into the potential mechanisms of cell death in the inflamed human joint.

• Authors: Matthew Whiteman, Siew Hwa Chu, Jia Ling Siau, Peter Rose, Kanaga Sabapathy, Jan-Thorsten Schantz, Nam Sang Cheung, Jeremy P E Spencer, Jeffrey S Armstrong
• Journal: Cellular signalling (Cell Signal) Vol. 19 Issue 4 Pg. 705-14 (Apr 2007) ISSN: 0898-6568 [Print] England
• PMID: 17107772 (Publication Type: Journal Article)

Chemical References

• Apoptosis Inducing Factor
• Inflammation Mediators
• Oxidants
• bcl-2-Associated X Protein
• Hypochlorous Acid
• Endodeoxyribonucleases
• endonuclease G
• Caspases

Topics

• Apoptosis Inducing Factor (metabolism)
• Caspases (metabolism)
• Catalysis (drug effects)
• Cell Death (drug effects)
• Chondrocytes (cytology, drug effects)
• Endodeoxyribonucleases (metabolism)
• Enzyme Activation (drug effects)
• Humans
• Hypochlorous Acid (pharmacology)
• Inflammation Mediators (pharmacology)
• Membrane Potential, Mitochondrial (drug effects)
• Mitochondrial Membranes (drug effects)
• Models, Biological
• Oxidants (pharmacology)
• Permeability (drug effects)
• Protein Conformation (drug effects)
• Protein Transport (drug effects)
• bcl-2-Associated X Protein (chemistry, metabolism)