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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 activation of ERK is involved in the CD147-induced expression of MMP-9 and suppression of p38 and JNK activity augments the MMP-9 expression. (A) THP-1 cells were stimulated with 1µg/ml LPS or immobilized anti-CD147 mAb (10µg/ml) in the presence or absence or indicated amounts of PD98059 (PD), U0126 (U), SB203580 (SB), JNK inhibitor (JNK-I), or 0.2% DMSO as a vehicle control (VC). Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. Numbers below each lane represent the band intensity which was normalized with band intensity of sample treated with only anti-CD147 in each gel. (B) THP-1 cells were stimulated with immobilized anti-CD147 mAb (10µg/ml). Cell lysates were collected at indicated times and the levels of phospho-ERK or ERK were analyzed using Western blot analysis. (C) THP-1 cells were stimulated with 0.1µM of CypA in the presence or absence of indicated amounts of PD98059 (PD), U0126 (U), SB203580 (SB), JNK inhibitor (JNK-I), negative control for JNK-I [J(-)], or 0.2% DMSO as a vehicle control (VC). Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. (D) The bar graph shows the MMP-9 band intensities of each lane in panel (C) that was normalized with band intensity of sample treated with only CypA in each gel.
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Figure 6: The activation of ERK is involved in the CD147-induced expression of MMP-9 and suppression of p38 and JNK activity augments the MMP-9 expression. (A) THP-1 cells were stimulated with 1µg/ml LPS or immobilized anti-CD147 mAb (10µg/ml) in the presence or absence or indicated amounts of PD98059 (PD), U0126 (U), SB203580 (SB), JNK inhibitor (JNK-I), or 0.2% DMSO as a vehicle control (VC). Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. Numbers below each lane represent the band intensity which was normalized with band intensity of sample treated with only anti-CD147 in each gel. (B) THP-1 cells were stimulated with immobilized anti-CD147 mAb (10µg/ml). Cell lysates were collected at indicated times and the levels of phospho-ERK or ERK were analyzed using Western blot analysis. (C) THP-1 cells were stimulated with 0.1µM of CypA in the presence or absence of indicated amounts of PD98059 (PD), U0126 (U), SB203580 (SB), JNK inhibitor (JNK-I), negative control for JNK-I [J(-)], or 0.2% DMSO as a vehicle control (VC). Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. (D) The bar graph shows the MMP-9 band intensities of each lane in panel (C) that was normalized with band intensity of sample treated with only CypA in each gel.

Mentions: CypA has been reported to induce the activation of ERK1/2 in various cell types such as cancer cells, neurons, and leukocytes (21,33-35). In order to verify the involvement of MAPKs for the expression of MMP-9 in cells stimulated with CD147, the assay was performed in the presence of MAPK inhibitors. Inhibitors of ERK MAPK (U0126 and PD98059) blocked the secretion of MMP-9 in a dose-dependent manner (Fig. 6A, note the numbers below each lane). The involvement of ERK in CD147-mediated signaling was further confirmed by detecting the phosphorylation of ERK using Western blot analysis in cells stimulated with anti-CD147 mAb (Fig. 6B). Interestingly, treatment with inhibitors of p38 and JNK MAPK slightly induced MMP-9 expression (Fig. 6A, note the members below each lane which represent the relative band intensity). The involvement of MAPKs was further confirmed in THP-1 cells stimulated with CypA. As shown in Fig. 6C, ERK inhibitors suppressed MMP-9 expression in a dose dependent manner, while the inhibitors of p38 and JNK enhanced the secretion of MMP-9 (Fig. 6C and 6D). The enhancement of ERK signaling by the suppression of p38 and/or JNK has been previously reported: the presence of p38 inhibitor caused an increase in basal phosphorylation level of ERK, which resulted in the enhanced ERK-mediated signaling and cellular responses in THP-1 cells (36,37). Similar enhancement of ERK phosphorylation in the presence of p38 and JNK inhibitors is likely the cause of the observed phenomenon. The molecular mechanism underlying this suppression of ERK activity by p38 and JNK is not known.


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 activation of ERK is involved in the CD147-induced expression of MMP-9 and suppression of p38 and JNK activity augments the MMP-9 expression. (A) THP-1 cells were stimulated with 1µg/ml LPS or immobilized anti-CD147 mAb (10µg/ml) in the presence or absence or indicated amounts of PD98059 (PD), U0126 (U), SB203580 (SB), JNK inhibitor (JNK-I), or 0.2% DMSO as a vehicle control (VC). Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. Numbers below each lane represent the band intensity which was normalized with band intensity of sample treated with only anti-CD147 in each gel. (B) THP-1 cells were stimulated with immobilized anti-CD147 mAb (10µg/ml). Cell lysates were collected at indicated times and the levels of phospho-ERK or ERK were analyzed using Western blot analysis. (C) THP-1 cells were stimulated with 0.1µM of CypA in the presence or absence of indicated amounts of PD98059 (PD), U0126 (U), SB203580 (SB), JNK inhibitor (JNK-I), negative control for JNK-I [J(-)], or 0.2% DMSO as a vehicle control (VC). Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. (D) The bar graph shows the MMP-9 band intensities of each lane in panel (C) that was normalized with band intensity of sample treated with only CypA in each gel.
© Copyright Policy - open-access
Related In: Results  -  Collection

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Show All Figures
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Figure 6: The activation of ERK is involved in the CD147-induced expression of MMP-9 and suppression of p38 and JNK activity augments the MMP-9 expression. (A) THP-1 cells were stimulated with 1µg/ml LPS or immobilized anti-CD147 mAb (10µg/ml) in the presence or absence or indicated amounts of PD98059 (PD), U0126 (U), SB203580 (SB), JNK inhibitor (JNK-I), or 0.2% DMSO as a vehicle control (VC). Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. Numbers below each lane represent the band intensity which was normalized with band intensity of sample treated with only anti-CD147 in each gel. (B) THP-1 cells were stimulated with immobilized anti-CD147 mAb (10µg/ml). Cell lysates were collected at indicated times and the levels of phospho-ERK or ERK were analyzed using Western blot analysis. (C) THP-1 cells were stimulated with 0.1µM of CypA in the presence or absence of indicated amounts of PD98059 (PD), U0126 (U), SB203580 (SB), JNK inhibitor (JNK-I), negative control for JNK-I [J(-)], or 0.2% DMSO as a vehicle control (VC). Culture supernatants were collected 24 hr after activation and subjected to gelatin zymogram. (D) The bar graph shows the MMP-9 band intensities of each lane in panel (C) that was normalized with band intensity of sample treated with only CypA in each gel.
Mentions: CypA has been reported to induce the activation of ERK1/2 in various cell types such as cancer cells, neurons, and leukocytes (21,33-35). In order to verify the involvement of MAPKs for the expression of MMP-9 in cells stimulated with CD147, the assay was performed in the presence of MAPK inhibitors. Inhibitors of ERK MAPK (U0126 and PD98059) blocked the secretion of MMP-9 in a dose-dependent manner (Fig. 6A, note the numbers below each lane). The involvement of ERK in CD147-mediated signaling was further confirmed by detecting the phosphorylation of ERK using Western blot analysis in cells stimulated with anti-CD147 mAb (Fig. 6B). Interestingly, treatment with inhibitors of p38 and JNK MAPK slightly induced MMP-9 expression (Fig. 6A, note the members below each lane which represent the relative band intensity). The involvement of MAPKs was further confirmed in THP-1 cells stimulated with CypA. As shown in Fig. 6C, ERK inhibitors suppressed MMP-9 expression in a dose dependent manner, while the inhibitors of p38 and JNK enhanced the secretion of MMP-9 (Fig. 6C and 6D). The enhancement of ERK signaling by the suppression of p38 and/or JNK has been previously reported: the presence of p38 inhibitor caused an increase in basal phosphorylation level of ERK, which resulted in the enhanced ERK-mediated signaling and cellular responses in THP-1 cells (36,37). Similar enhancement of ERK phosphorylation in the presence of p38 and JNK inhibitors is likely the cause of the observed phenomenon. The molecular mechanism underlying this suppression of ERK activity by p38 and JNK is not known.

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