TY - JOUR

T1 - Hypothiocyanous acid is a more potent inducer of apoptosis and protein thiol depletion in murine macrophage cells than hypochlorous acid or hypobromous acid

AU - Lloyd, Mitchell M

AU - van Reyk, David M

AU - Davies, Michael Jonathan

AU - Hawkins, Clare Louise

PY - 2008/9/1

Y1 - 2008/9/1

N2 - Hypohalous acids are generated by activated leucocytes, via the formation of H(2)O(2) and the release of peroxidase enzymes (myeloperoxidase and eosinophil peroxidase). These species are important bactericidal agents, but HOCl (hypochlorous acid) and HOBr (hypobromous acid) have also been implicated in tissue damage in a number of inflammatory diseases. HOSCN (hypothiocyanous acid; cyanosulfenic acid) is a milder, more thiol-specific, oxidant than HOCl or HOBr and as such may be a more potent inducer of cellular dysfunction due to selective targeting of critical thiol residues on proteins. In the present study, HOCl and HOBr are shown to react rapidly with macrophage (J774A.1) cells, resulting in a greater extent of cell lysis compared with HOSCN. However, HOSCN induces apoptosis and necrosis with greater efficacy, and at lower concentrations, than HOCl or HOBr. Apoptosis occurs in conjunction with an increased release of cytochrome c into the cytosol, but no associated increase in caspase activity. Similarly, apoptosis is observed on treating the cells in the presence of a caspase inhibitor, suggesting that it is mediated by a caspase-independent pathway. HOSCN oxidized protein thiols more efficiently than either HOCl or HOBr. The greater efficacy of HOSCN in inducing apoptosis is attributed to selective damage to critical mitochondrial membrane protein thiol groups, resulting in increased permeability and subsequent leakage of cytochrome c into the cytosol. This induction of damage by HOSCN may be of critical importance in people with elevated levels of SCN(-) (thiocyanate ions) arising from cigarette smoking, and plays a role in the pathologies associated with this biological insult.

AB - Hypohalous acids are generated by activated leucocytes, via the formation of H(2)O(2) and the release of peroxidase enzymes (myeloperoxidase and eosinophil peroxidase). These species are important bactericidal agents, but HOCl (hypochlorous acid) and HOBr (hypobromous acid) have also been implicated in tissue damage in a number of inflammatory diseases. HOSCN (hypothiocyanous acid; cyanosulfenic acid) is a milder, more thiol-specific, oxidant than HOCl or HOBr and as such may be a more potent inducer of cellular dysfunction due to selective targeting of critical thiol residues on proteins. In the present study, HOCl and HOBr are shown to react rapidly with macrophage (J774A.1) cells, resulting in a greater extent of cell lysis compared with HOSCN. However, HOSCN induces apoptosis and necrosis with greater efficacy, and at lower concentrations, than HOCl or HOBr. Apoptosis occurs in conjunction with an increased release of cytochrome c into the cytosol, but no associated increase in caspase activity. Similarly, apoptosis is observed on treating the cells in the presence of a caspase inhibitor, suggesting that it is mediated by a caspase-independent pathway. HOSCN oxidized protein thiols more efficiently than either HOCl or HOBr. The greater efficacy of HOSCN in inducing apoptosis is attributed to selective damage to critical mitochondrial membrane protein thiol groups, resulting in increased permeability and subsequent leakage of cytochrome c into the cytosol. This induction of damage by HOSCN may be of critical importance in people with elevated levels of SCN(-) (thiocyanate ions) arising from cigarette smoking, and plays a role in the pathologies associated with this biological insult.

KW - Animals

KW - Apoptosis

KW - Bromates

KW - Caspases

KW - Cell Line

KW - Cytochromes c

KW - Glutathione

KW - Glutathione Disulfide

KW - Hypochlorous Acid

KW - Macrophages

KW - Mice

KW - Necrosis

KW - Oxidation-Reduction

KW - Sulfhydryl Compounds

KW - Thiocyanates

U2 - 10.1042/BJ20080468

DO - 10.1042/BJ20080468

M3 - Journal article

C2 - 18459943

VL - 414

SP - 271

EP - 280

JO - Biochemical Journal

JF - Biochemical Journal

SN - 0264-6021

IS - 2

ER -