Author/Authors :
Kawata، Jin نويسنده Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan , , Yamaguchi، Rui نويسنده Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan , , Yamamoto، Takatoshi نويسنده Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan , , Ishimaru، Yasuji نويسنده Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan , , Sakamoto، Arisa نويسنده Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan * , , Aoki، Manabu نويسنده Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan , , Kitano، Masafumi نويسنده Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan , , Umehashi، Misako نويسنده Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan , , Hirose، Eiji نويسنده Graduate School of Medical Science, Kumamoto Health Science University, Kumamoto, Japan , , Yamaguchi، Yasuo نويسنده ,
Abstract :
Objective: Neutrophils have an important role in the rapid innate immune response,
and the release or active secretion of elastase from neutrophils is linked to various
inflammatory responses. Purpose of this study was to determine how the human
neutrophil elastase affects the interleukin-10 (IL-10) response in peripheral blood
mononuclear cells (PBMC).
Materials and Methods: In this prospective study, changes in IL-10 messenger RNA
(mRNA) and protein expression levels in monocytes derived from human PBMCs
were investigated after stimulation with human neutrophil elastase (HNE). A set of
inhibitors was used for examining the pathways for IL-10 production induced by HNE.
Results: Reverse transcription polymerase chain reaction (RT-PCR) showed that
stimulation with HNE upregulated IL-10 mRNA expression by monocytes, while the
enzyme-linked immunosorbent assay (ELISA) revealed an increase of IL-10 protein
level in the culture medium. A phospholipase C inhibitor (U73122) partially blunted
the induction of IL-10 mRNA expression by HNE, while IL-10 mRNA expression
was significantly reduced by a protein kinase C (PKC) inhibitor (Rottlerin). A calcium
chelator (3,4,5-trimethoxybenzoic acid 8-(diethylamino)octyl ester: TMB-8) inhibited
the response of IL-10 mRNA to stimulation by HNE. In addition, pretreatment with
a broad-spectrum PKC inhibitor (Ro-318425) partly blocked the response to HNE.
Finally, an inhibitor of PKC theta/delta abolished the increased level of IL-10 mRNA
expression.
Conclusion: These results indicate that HNE mainly upregulates IL-10 mRNA expression
and protein production in moncytes via a novel PKC theta/delta, although
partially via the conventional PKC pathway.
