Suppressive effects of sivelestat on interleukin 8 and TNF-α production from LPS-stimulated granulocytes in whole blood culture.
OBJECTIVE
The goal of the study was to examine the effects of sivelestat sodium hydrate (sivelestat), a neutrophil elastase inhibitor, on production of cytokines in granulocytes and monocytes, using flow cytometry after cytokine staining in whole blood culture.


METHODS
Blood samples were collected from healthy volunteers. Vehicle (control group), lipopolysaccharide (LPS) (LPS group), or LPS + sivelestat (sivelestat group) were added to the whole blood, followed by addition of a protein transport inhibitor in each group. After incubation, staining for cytokines retained in the cells was performed by addition of an anti-interleukin 8 (IL-8) or anti-tumor necrosis factor-α (TNF-α) antibody. The cells were then analyzed using flow cytometry.


RESULTS
Granulocytic production of IL-8 induced by 1 ng/ml LPS was significantly (P < 0.05) inhibited by treatment with 1 μg/ml sivelestat, and upregulation of IL-8 by 10 ng/ml LPS was also significantly (P < 0.05) suppressed by 1 and 10 μg/ml sivelestat. Addition of 10 or 100 μg/ml sivelestat significantly (P < 0.05) inhibited the production of TNF-α from granulocytes induced by 10 ng/ml LPS. Sivelestat did not significantly inhibit LPS-induced monocytic production of TNF-α and IL-8.


CONCLUSIONS
Suppression of granulocytic production of IL-8 and TNF-α by sivelestat suggests that this drug may be useful for treatment of morbid conditions involving IL-8 and TNF-α at onset.
Department of Critical Care Medicine, Iwate Medical University, 19-1 Uchimaru, Morioka 020-8505, Japan. ac100277@akita-city-hp.jp
Shigehiro
S
Shibata
Gaku
G
Takahashi
Nobuki
N
Shioya
Katsuya
K
Inada
Shigeatsu
S
Endo
D055371:Acute Lung Injury / Q000188:drug therapy / Q000503:physiopathology; D005434:Flow Cytometry; D005998:Glycine / Q000031:analogs & derivatives* / Q000494:pharmacology; D006098:Granulocytes / Q000187:drug effects / Q000378:metabolism*; D006801:Humans; D066298:In Vitro Techniques; D016209:Interleukin-8 / Q000096:biosynthesis*; D008070:Lipopolysaccharides / Q000494:pharmacology*; D009000:Monocytes / Q000187:drug effects / Q000378:metabolism; D053491:Proteinase Inhibitory Proteins, Secretory / Q000494:pharmacology*; D013449:Sulfonamides / Q000494:pharmacology*; D014409:Tumor Necrosis Factor-alpha / Q000096:biosynthesis*
D016209:Interleukin-8; D008070:Lipopolysaccharides; D053491:Proteinase Inhibitory Proteins, Secretory; D013449:Sulfonamides; D014409:Tumor Necrosis Factor-alpha; C069195:sivelestat; D005998:Glycine