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The Stimulation of CD147 Induces MMP-9 Expression through ERK and NF-kappaB in Macrophages: Implication for Atherosclerosis.

Kim JY, Kim WJ, Kim H, Suk K, Lee WH - Immune Netw (2009)

Bottom Line: Recent observations showing the expression of CD147 in leukocytes indicate that this molecule may have roles in inflammation.Staining of both CD147 and CypA was detected in endothelial cell layers facing the lumen and macrophage-rich areas.These results suggest that CD147 mediates the inflammatory activation of macrophages that leads to the induction of MMP-9 expression, which could play a role in the pathogenesis of inflammatory diseases such as atherosclerosis.

View Article: PubMed Central - PubMed

Affiliation: The School of Life Sciences and Biotechnology, Kyungpook National University, Daegu 702-701, Korea.

ABSTRACT

Background: CD147, as a cellular receptor for cyclophilin A (CypA), is a multifunctional protein involved in tumor invasion, inflammation, tissue remodeling, neural function, and reproduction. Recent observations showing the expression of CD147 in leukocytes indicate that this molecule may have roles in inflammation.

Methods: In order to investigate the role of CD147 and its ligand in the pathogenesis of atherosclerosis, human atherosclerotic plaques were analyzed for the expression pattern of CD147 and CypA. The cellular responses and signaling molecules activated by the stimulation of CD147 were then investigated in the human macrophage cell line, THP-1, which expresses high basal level of CD147 on the cell surface.

Results: Staining of both CD147 and CypA was detected in endothelial cell layers facing the lumen and macrophage-rich areas. Stimulation of CD147 with its specific monoclonal antibody induced the expression of matrix metalloproteinase (MMP)-9 in THP-1 cells and it was suppressed by inhibitors of both ERK and NF-kappaB. Accordingly, the stimulation of CD147 was observed to induce phosphorylation of ERK, phosphorylation-associated degradation of IkappaB, and nuclear translocation of NF-kappaB p65 and p50 subunits.

Conclusion: These results suggest that CD147 mediates the inflammatory activation of macrophages that leads to the induction of MMP-9 expression, which could play a role in the pathogenesis of inflammatory diseases such as atherosclerosis.

No MeSH data available.


Related in: MedlinePlus

The stimulation of CD147 induces nuclear translocation of NF-κB p65 and p50 subunits. (A) THP-1 cells were stimulated with 1µg/ml of LPS or anti-CD147 mAb that had been immobilized at a concentration of 10µg/ml. Nuclear extracts were collected after indicated times and the Western blot analysis was performed using p65- or TFIIB-specific antibodies. TFIIB was used as a loading control for nuclear extracts. (B, C) THP-1 cells were stimulated with LPS, anti-CD147 mAb, or isotype-matching mouse IgG (mIgG) for 2 hr and analyzed with immunofluorescence analysis using an antibody specific for NF-κB p50 subunit. (B) shows representative pictures of cells in each samples and (C) shows the percentage of cells that had NF-κB p50 subunit at their nucleus.
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Figure 4: The stimulation of CD147 induces nuclear translocation of NF-κB p65 and p50 subunits. (A) THP-1 cells were stimulated with 1µg/ml of LPS or anti-CD147 mAb that had been immobilized at a concentration of 10µg/ml. Nuclear extracts were collected after indicated times and the Western blot analysis was performed using p65- or TFIIB-specific antibodies. TFIIB was used as a loading control for nuclear extracts. (B, C) THP-1 cells were stimulated with LPS, anti-CD147 mAb, or isotype-matching mouse IgG (mIgG) for 2 hr and analyzed with immunofluorescence analysis using an antibody specific for NF-κB p50 subunit. (B) shows representative pictures of cells in each samples and (C) shows the percentage of cells that had NF-κB p50 subunit at their nucleus.

Mentions: The expression of MMP-9 in macrophage requires the activation of NF-κB in macrophages. NF-κB, a heterodimer of p65 and p50, stays in cytoplasm in its inactive status in association with IκB. When the activation signal(s) is transmitted, IκB become phosphorylated and, as a result, degraded by proteasome. The free NF-κB heterodimer then translocates into the nucleus. In order to analyze the requirement of NF-κB activation in the CD147-induced expression of MMP-9, immunohistochemistry and Western blot analysis was performed using p65 or p50 specific antibodies. As shown in Fig. 4A, the level of nuclear p65 was increased 30 to 60 min after activation with anti-CD147 mAb. In accordance with this data, nuclear translocation of NF-κB p50 subunit was also detected in cells stimulated with anti-CD147 mAb (Fig. 4B and 4C).


The Stimulation of CD147 Induces MMP-9 Expression through ERK and NF-kappaB in Macrophages: Implication for Atherosclerosis.

Kim JY, Kim WJ, Kim H, Suk K, Lee WH - Immune Netw (2009)

The stimulation of CD147 induces nuclear translocation of NF-κB p65 and p50 subunits. (A) THP-1 cells were stimulated with 1µg/ml of LPS or anti-CD147 mAb that had been immobilized at a concentration of 10µg/ml. Nuclear extracts were collected after indicated times and the Western blot analysis was performed using p65- or TFIIB-specific antibodies. TFIIB was used as a loading control for nuclear extracts. (B, C) THP-1 cells were stimulated with LPS, anti-CD147 mAb, or isotype-matching mouse IgG (mIgG) for 2 hr and analyzed with immunofluorescence analysis using an antibody specific for NF-κB p50 subunit. (B) shows representative pictures of cells in each samples and (C) shows the percentage of cells that had NF-κB p50 subunit at their nucleus.
© Copyright Policy - open-access
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC2803300&req=5

Figure 4: The stimulation of CD147 induces nuclear translocation of NF-κB p65 and p50 subunits. (A) THP-1 cells were stimulated with 1µg/ml of LPS or anti-CD147 mAb that had been immobilized at a concentration of 10µg/ml. Nuclear extracts were collected after indicated times and the Western blot analysis was performed using p65- or TFIIB-specific antibodies. TFIIB was used as a loading control for nuclear extracts. (B, C) THP-1 cells were stimulated with LPS, anti-CD147 mAb, or isotype-matching mouse IgG (mIgG) for 2 hr and analyzed with immunofluorescence analysis using an antibody specific for NF-κB p50 subunit. (B) shows representative pictures of cells in each samples and (C) shows the percentage of cells that had NF-κB p50 subunit at their nucleus.
Mentions: The expression of MMP-9 in macrophage requires the activation of NF-κB in macrophages. NF-κB, a heterodimer of p65 and p50, stays in cytoplasm in its inactive status in association with IκB. When the activation signal(s) is transmitted, IκB become phosphorylated and, as a result, degraded by proteasome. The free NF-κB heterodimer then translocates into the nucleus. In order to analyze the requirement of NF-κB activation in the CD147-induced expression of MMP-9, immunohistochemistry and Western blot analysis was performed using p65 or p50 specific antibodies. As shown in Fig. 4A, the level of nuclear p65 was increased 30 to 60 min after activation with anti-CD147 mAb. In accordance with this data, nuclear translocation of NF-κB p50 subunit was also detected in cells stimulated with anti-CD147 mAb (Fig. 4B and 4C).

Bottom Line: Recent observations showing the expression of CD147 in leukocytes indicate that this molecule may have roles in inflammation.Staining of both CD147 and CypA was detected in endothelial cell layers facing the lumen and macrophage-rich areas.These results suggest that CD147 mediates the inflammatory activation of macrophages that leads to the induction of MMP-9 expression, which could play a role in the pathogenesis of inflammatory diseases such as atherosclerosis.

View Article: PubMed Central - PubMed

Affiliation: The School of Life Sciences and Biotechnology, Kyungpook National University, Daegu 702-701, Korea.

ABSTRACT

Background: CD147, as a cellular receptor for cyclophilin A (CypA), is a multifunctional protein involved in tumor invasion, inflammation, tissue remodeling, neural function, and reproduction. Recent observations showing the expression of CD147 in leukocytes indicate that this molecule may have roles in inflammation.

Methods: In order to investigate the role of CD147 and its ligand in the pathogenesis of atherosclerosis, human atherosclerotic plaques were analyzed for the expression pattern of CD147 and CypA. The cellular responses and signaling molecules activated by the stimulation of CD147 were then investigated in the human macrophage cell line, THP-1, which expresses high basal level of CD147 on the cell surface.

Results: Staining of both CD147 and CypA was detected in endothelial cell layers facing the lumen and macrophage-rich areas. Stimulation of CD147 with its specific monoclonal antibody induced the expression of matrix metalloproteinase (MMP)-9 in THP-1 cells and it was suppressed by inhibitors of both ERK and NF-kappaB. Accordingly, the stimulation of CD147 was observed to induce phosphorylation of ERK, phosphorylation-associated degradation of IkappaB, and nuclear translocation of NF-kappaB p65 and p50 subunits.

Conclusion: These results suggest that CD147 mediates the inflammatory activation of macrophages that leads to the induction of MMP-9 expression, which could play a role in the pathogenesis of inflammatory diseases such as atherosclerosis.

No MeSH data available.


Related in: MedlinePlus