J Korean Acad Periodontol.  2005 Jun;35(2):427-436. 10.5051/jkape.2005.35.2.427.

Effect of CD14, Toll-like receptors, cytoskeletal inhibitors and NF-kappaB inhibitor on MMP-8 release from human neutrophils induced by E. coli lipopolysaccharides

• Yang SM
• Kim TI
• Seol YJ
• Lee YM
• Ku Y
• Chung CP
• Han SB
• Rhyu IC

• KMID: 1999769
• DOI: http://doi.org/10.5051/jkape.2005.35.2.427

Abstract

OBJECTIVE
MMP-8 is a neutrophil enzyme and its level increases in some inflammatory diseases, including periodontal disease. We knew that the lipopolysaccharide of E.coli(E-LPS) induced MMP-8 release from human neutrophils. E-LPS is known to induce the production and release of inflammatory cytokines through CD14, Toll-like receptor(TLR). In the present study, we investigated whether MMP-8 release by E-LPS is induced via CD14-TLR pathway and the cellular mechanism of MMP-8 release in human neutrophils. MATERIAL AND METHODS: Human neutrophils were isolated from the peripheral blood of healthy donors and pre-incubated in medium containing antibodies against CD14, anti-TLR2 and anti-TLR4 or several inhibitors of microtubules and microfilaments and then incubated with E-LPS. The cells were treated TPCK and E-LPS simultaneously. The MMP-8amount in the culture medium was determined using ELISA.
RESULTS
E-LPS increased MMP-8release from neutrophils and its induction was inhibited by anti- CD14 and anti-TLR4 but not by anti-TLR2 antibodies. The inhibitors of micro- tubule and microfilament polymerization significantly decreased E-LPS-induced MMP- 8release. TPCK inhibited E-LPS-induced MMP-8 release.
CONCLUSION
These results suggest that MMP-8 release is induced by E-LPS via the CD14-TLR4 signal pathway in human neutrophils and may be depedent on microtubule and microfilament systems and NF-kappaB pathway.

Keyword

Human Neutrophil; CD14; TLRs; Cytoskeletal Inhibitors; NF-B inhibitor; MMP-8

MeSH Terms

Actin Cytoskeleton
Antibodies
Cytokines
Enzyme-Linked Immunosorbent Assay
Humans*
Lipopolysaccharides*
Microtubules
Neutrophils*
NF-kappa B*
Periodontal Diseases
Polymerization
Polymers
Signal Transduction
Tissue Donors
Toll-Like Receptors*
Tosylphenylalanyl Chloromethyl Ketone
Antibodies
Cytokines
Lipopolysaccharides
NF-kappa B
Polymers
Toll-Like Receptors
Tosylphenylalanyl Chloromethyl Ketone