FSL-1 Induces MMP-9 Production through TLR-2 and NF-κB /AP-1 Signaling Pathways in Monocytic THP-1 Cells

Abstract

Background: Matrix metalloproteinase-9 (MMP-9) is known to be implicated in the pathogenesis of many inflammatory disorders. FSL-1 (fibroblast-stimulating lipopeptide-1) induces cytokine production by monocytes/macrophages. However, it is unclear whether FSL-1 is also able to induce MMP-9 production. Herein, we determined whether FSL-1 could induce MMP-9 production, and if so, which signal transduction pathway(s) were involved. Methods: MMP-9 expression was assessed with real-time qPCR and ELISA. Signaling pathways were studied by using THP1-XBlue™ cells, THP1-XBlue™-defMyD cells, anti-TLR2 mAb and pharmacological inhibitors. Phospho and total proteins were determined by Western blotting. Results: FSL-1 induces MMP-9 expression (P<0.001) at both mRNA and protein levels in human monocytic THP-1 cells. Elevated activity (P<0.001) of NF-κB/AP-1 was also observed in FSL-1-treated THP-1 cells. FSL-1-induced MMP-9 secretion was markedly suppressed either by neutralizing anti-TLR-2 antibody or by inhibiting clathrin-dependent endocytosis. Furthermore, MyD88^-/- THP-1 cells did not express MMP-9 in response to FSL-1 treatment. By small interfering RNA-mediated knockdown, we also show that FSL-1-induced up-regulation of MMP-9 requires MyD88. Pre-treatment of THP-1 cells with inhibitors of JNK (SP600125), MEK/ERK (U0126; PD98056; XMD 8-92), p38 MAPK (SB203580) and NF-κB (BAY11-7085, Triptolide, Resveratrol) significantly suppressed (P<0.05) MMP-9 gene expression and NF-κB/AP-1 transcription factors activity. Conclusion: These findings provide the first evidence that FSL-1 induces TLR-2-dependent MMP-9 gene expression which requires the recruitment of MyD88 and leads to activation of MEK1/2 /ERK 1/2, MEK5/ERK5, JNK, p38 MAPK and NF-κB/AP-1.

MMP2 and MMP9 are Associated with Apical Periodontitis Progression and Might be Modulated by TLR2 and MyD88

“…MMP9 is also essential for initiating the osteoclastic resorption process by removing the collagenous layer from the bone surface before demineralization can start (14). An interaction between TLR2 and MyD88 with MMP9 has also been demonstrated in monocytic cells (25). In our study, we noted that the expression of MMP9 in both KO mice were different from the WT.…”

Pam3CSK4 Induces MMP-9 Expression in Human Monocytic THP-1 Cells

“…Bacterial components such as LPS, lipopeptides, lipoteichoic acids (LTAs), and peptidoglycan stimulate the production of cytokines and other inflammatory mediators [35,36]. FSL-1 and HKLM were reported to induce MMP-9 production [23,24,37]. Herein, we show that Pam3CSK4 induces MMP-9 production in monocytic cells.…”

“…Pam3CSK4-induced, TLR2-mediated MMP-9 expression in THP-1 cells involves clathrin-dependent endocytosis. THP-1 cell cultures were treated with pharmacological endocytosis inhibitors; chlorpromazine (an inhibitor of clathrin-dependent endocytosis; 10uM) and Methyl-β-cyclodextrin (MBCD; an inhibitor clathrin-independent endocytosis; 2mM) for 1hr prior to treatment with Pam3CSK4 for 24hr [23]. Cells and conditioned media were collected and used for MMP-9 expression.…”

Acetylcholine Inhibits LPS-Induced MMP-9 Production and Cell Migration via the a7 nAChR-JAK2/STAT3 Pathway in RAW264.7 Cells

“…MMP-9 is tightly regulated at several levels, including transcription, translation, proenzyme activation, and the tissue inhibitor of metalloproteinases (TIMPs); the transcriptional regulation is particularly important [6,7,10]. Several transcription factors have been shown to bind to the promoter region of the MMP-9 gene, including nuclear factor-κB (NF-κB) and activating protein 1 (AP-1), which leads to MMP-9 transcription [11,12]. In contrast, signal transducer and activator of transcription 3 (STAT3), a potential anti-inflammatory mediator, can inhibit MMP-9 expression [13].…”

“…In contrast, signal transducer and activator of transcription 3 (STAT3), a potential anti-inflammatory mediator, can inhibit MMP-9 expression [13]. Other studies have demonstrated that activation of p38 mitogen-activated protein kinase (MAPK) and extracellular regulated protein kinases 1/2 (ERK1/2) up-regulated MMP-9 expression [12,14,15], while JNK (c-Jun N-Terminal kinase) phosphorylation reduced MMP-9 expression [15,16]. Thus, upstream regulators of MMP-9, such as STAT3 and NF-κB, may be utilized as therapeutic targets and can provide new insights into the treatment of many inflammatory diseases.…”