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2018 ; 9
(ä): 359
Nephropedia Template TP
Front Immunol
2018[]; 9
(ä): 359
PMID29593709
show ga
Tonicity of saline (NaCl) is important in regulating cellular functions and
homeostasis. Hypertonic saline is administered to treat many inflammatory
diseases, including cystic fibrosis. Excess neutrophil extracellular trap (NET)
formation, or NETosis, is associated with many pathological conditions including
chronic inflammation. Despite the known therapeutic benefits of hypertonic
saline, its underlying mechanisms are not clearly understood. Therefore, we aimed
to elucidate the effects of hypertonic saline in modulating NETosis. For this
purpose, we purified human neutrophils and induced NETosis using agonists such as
diacylglycerol mimetic phorbol myristate acetate (PMA), Gram-negative bacterial
cell wall component lipopolysaccharide (LPS), calcium ionophores (A23187 and
ionomycin from Streptomyces conglobatus), and bacteria (Pseudomonas aeruginosa
and Staphylococcus aureus). We then analyzed neutrophils and NETs using Sytox
green assay, immunostaining of NET components and apoptosis markers, confocal
microscopy, and pH sensing reagents. This study found that hypertonic NaCl
suppresses nicotinamide adenine dinucleotide phosphate oxidase (NADPH2 or
NOX2)-dependent NETosis induced by agonists PMA, Escherichia coli LPS (0111:B4
and O128:B12), and P. aeruginosa. Hypertonic saline also suppresses LPS- and PMA-
induced reactive oxygen species production. It was determined that supplementing
H(2)O(2) reverses the suppressive effect of hypertonic saline on NOX2-dependent
NETosis. Many of the aforementioned suppressive effects were observed in the
presence of equimolar concentrations of choline chloride and osmolytes
(d-mannitol and d-sorbitol). This suggests that the mechanism by which hypertonic
saline suppresses NOX2-dependent NETosis is via neutrophil dehydration.
Hypertonic NaCl does not significantly alter the intracellular pH of neutrophils.
We found that hypertonic NaCl induces apoptosis while suppressing NOX2-dependent
NETosis. In contrast, hypertonic solutions do not suppress NOX2-independent
NETosis. Although hypertonic saline partially suppresses ionomycin-induced
NETosis, it enhances A23187-induced NETosis, and it does not alter S.
aureus-induced NETosis. Overall, this study determined that hypertonic saline
suppresses NOX2-dependent NETosis induced by several agonists; in contrast, it
has variable effects on neutrophil death induced by NOX2-independent NETosis
agonists. These findings are important in understanding the regulation of NETosis
and apoptosis in neutrophils.