HOCl causes necrotic cell death in human monocyte derived macrophages through calcium dependent calpain activation.
The abundance of dead macrophages in close proximity to HOCl-modified proteins in advanced atherosclerotic plaques implicates HOCl in the killing of macrophages and the formation of the necrotic core region. The mechanism of HOCl mediated death of macrophages was unknown, so using human monocyte derived macrophages (HMDM) we here have shown that HOCl causes a rapid necrotic cell death characterized by loss of MTT reduction, cellular ATP and cell lysis without caspase-3 activation in HMDM cells. The HOCl causes a rise in cytosolic calcium level via the plasma membrane L- and T-type calcium channels and endoplasmic reticulum RyR channel. Blocking of the calcium channels or the addition of calpain inhibitors prevents the HOCl mediated loss of mitochondrial potential, lysosome failure and HMDM cell death. Blocking MPT-pore formation with cyclosporin A also prevents the loss of mitochondrial membrane potential, lysosomal destabilization and HMDM cell death. Blocking the calcium mitochondrial uniporter with ruthenium red also blocks the loss of mitochondrial potential but only at high concentrations. HOCl appears to cause HMDM cell death through destabilization of cytosolic calcium control resulting in the failure of both the mitochondria and lysosomes.
Free Radical Biochemistry Laboratory, School of Biological Sciences, University of Canterbury, Private Bag 4800, Christchurch, New Zealand.
Ya-ting Tina
YT
Yang
Matthew
M
Whiteman
Steven P
SP
Gieseg
D000001:Calcimycin / Q000494:pharmacology; D002118:Calcium / Q000378:metabolism*; D002121:Calcium Channel Blockers / Q000494:pharmacology; D061207:Calcium Ionophores / Q000494:pharmacology*; D002154:Calpain / Q000378:metabolism*; D053148:Caspase 3 / Q000378:metabolism; D016572:Cyclosporine / Q000494:pharmacology; D003620:Dantrolene / Q000494:pharmacology; D004791:Enzyme Inhibitors / Q000494:pharmacology; D005444:Flunarizine / Q000494:pharmacology; D006801:Humans; D006997:Hypochlorous Acid / Q000494:pharmacology*; D008247:Lysosomes / Q000187:drug effects; D008264:Macrophages / Q000187:drug effects* / Q000473:pathology* / Q000502:physiology; D053078:Membrane Potential, Mitochondrial / Q000187:drug effects; D009125:Muscle Relaxants, Central / Q000494:pharmacology; D009336:Necrosis; D009543:Nifedipine / Q000494:pharmacology; D014700:Verapamil / Q000494:pharmacology
D002121:Calcium Channel Blockers; D061207:Calcium Ionophores; D004791:Enzyme Inhibitors; D009125:Muscle Relaxants, Central; D000001:Calcimycin; D006997:Hypochlorous Acid; D016572:Cyclosporine; D014700:Verapamil; D002154:Calpain; D053148:Caspase 3; D003620:Dantrolene; D009543:Nifedipine; D005444:Flunarizine; D002118:Calcium