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