Clin Exp Otorhinolaryngol.  2014 Sep;7(3):175-180. 10.3342/ceo.2014.7.3.175.

Dexamethasone Inhibits Interleukin-1beta-Induced Matrix Metalloproteinase-9 Expression in Cochlear Cells

• Nam SI
• Kwon TK

• KMID: 1973466
• DOI: http://doi.org/10.3342/ceo.2014.7.3.175

Abstract

OBJECTIVES
To investigate the effect of interleukin (IL)-1beta on matrix metalloproteinase (MMP)-9 expression in cochlea and regulation of IL-1beta-mediated MMP-9 expression by dexamethasone and the molecular and signaling mechanisms involved.
METHODS
House ear institute-organ of Corti 1 (HEI-OC1) cells were used and exposed to IL-1beta with/without dexamethasone. Glucocorticoid receptor antagonist, RU486, was used to see the role of dexamethasone. PD98059 (an extracellular signal-regulated kinases [ERKs] inhibitor), SB203580 (a p38 mitogen-activated protein kinases [MAPK] inhibitor), SP600125 (a c-Jun N-terminal kinase [JNK] inhibitor) were also used to see the role of MAPKs signaling pathway(s) in IL-1beta-induced MMP-9 expression in HEI-OC1 cells. Reverse transcription-polymerase chain reaction and gelatin zymography were used to measure mRNA expression level of MMP-9 and activity of MMP-9, respectively.
RESULTS
Treatment with IL-1beta-induced the expression of MMP-9 in a dose- and time-dependent manner. IL-1beta (1 ng/mL)-induced MMP-9 expression was inhibited by dexamethasone. Interestingly, p38 MAPK inhibitor, SB203580, significantly inhibited IL-1beta-induced MMP-9 mRNA and MMP-9 activity. However, inhibition of JNKs and ERKs had no effect on the IL-1beta-induced MMP-9 expression.
CONCLUSION
These results suggest that the pro-inflammatory cytokine IL-1beta strongly induces MMP-9 expression via activation of p38 MAPK signaling pathway in HEI-OC1 cells and the induction was inhibited by dexamethasone.

Keyword

Interleukin-1beta; Matrix metalloproteinase 9; p38 Mitogen-activated protein kinases; Cochlea

MeSH Terms

Cochlea
Dexamethasone*
Ear
Extracellular Signal-Regulated MAP Kinases
Gelatin
Interleukin-1beta
Interleukins
JNK Mitogen-Activated Protein Kinases
Matrix Metalloproteinase 9*
Mifepristone
p38 Mitogen-Activated Protein Kinases
Receptors, Glucocorticoid
RNA, Messenger
Dexamethasone
Extracellular Signal-Regulated MAP Kinases
Gelatin
Interleukin-1beta
Interleukins
JNK Mitogen-Activated Protein Kinases
Matrix Metalloproteinase 9
Mifepristone
RNA, Messenger
Receptors, Glucocorticoid
p38 Mitogen-Activated Protein Kinases

Figure

• 

  Fig. 1 Effect of interleukin (IL)-1β on matrix metalloproteinase (MMP)-9 activity in house ear institute-organ of Corti 1 (HEI-OC1) cells. IL-1β increases MMP-9 activity and mRNA expression. (A, B) HEI-OC1 cells were treated with various concentrations of IL-1β for 20 hours. Conditional media were collected after 20 hours and gelatin zymography was performed (A). The MMP-9 mRNA expression levels were determined by reverse transcription-polymerase chain reaction (RT-PCR). The levels of actin were used as a loading control (B). (C, D) HEI-OC1 cells were treated with the 1 ng/mL IL-1β for the indicated time periods. Conditional media were collected after 20 hours and gelatin zymography was performed (C). The MMP-9 mRNA expression levels were determined by RT-PCR. The levels of actin were used as a loading control. The values in (B, D) represent the mean±SD from three independent samples. *P<0.001 compared to the control. The data represent three independent experiments.

• 

  Fig. 2 Effect of dexamethasone on interleukin (IL)-1β induced matrix metalloproteinase (MMP)-9 activity and MMP-9 mRNA expression. Dexamethasone inhibits IL-1β-induced MMP-9 activity and mRNA expression. House ear institute-organ of Corti 1 (HEI-OC1) cells were treated with the indicated concentrations of dexamethasone in the absence of presence of 1 ng/mL IL-1β for 20 hours. Conditional media were collected after 20 hours and gelatin zymography was performed (A). The MMP-9 mRNA expression levels were determined by reverse transcription-polymerase chain reaction. The levels of actin were used as a loading control. The values in (B) represent the mean±SD from three independent samples. *P<0.001 compared to IL-1β. The data represent three independent experiments.

• 

  Fig. 3 Effect of RU486 on inhibitory effect of dexamethasone in interleukin (IL)-1β-induced matrix metalloproteinase (MMP)-9 activity and mRNA expression. RU486 blocks inhibitory effect of dexamethasone in IL-1β-induced MMP-9 activity and mRNA expression. House ear institute-organ of Corti 1 (HEI-OC1) cells were pretreated with RU486 (10 µM) for 30 minutes, and then treated with the 1 ng/mL IL-1β in the absence or presence of 1 µM dexamethasone (Dex) for 20 hours. Conditional media were collected and gelatin zymography was performed (A). The MMP-9 mRNA expression levels were determined by reverse transcription-polymerase chain reaction. The levels of actin were used as a loading control. The values in (B) represent the mean±SD from three independent samples. *P<0.001 compared to IL-1β plus Dex. The data represent three independent experiments.

• 

  Fig. 4 Effects of N-acetylcysteine (NAC) on interleukin (IL)-1β-induced matrix metalloproteinase (MMP)-9 activity and mRNA expression. Reactive oxygen species signaling is not associated with IL-1β-indcued MMP-9 activity and mRNA expression. House ear institute-organ of Corti 1 (HEI-OC1) cells were pretreated with NAC (5 mM) for 30 minutes, and then added 1 ng/mL IL-1β for 20 hours. Conditional media were collected and gelatin zymography was performed (A). The MMP-9 mRNA expression levels were determined by reverse transcription-polymerase chain reaction. The levels of actin were used as a loading control. The values in (B) represent the mean±SD from three independent samples. The data represent three independent experiments.

• 

  Fig. 5 The p38 mitogen-activated protein kinases (MAPK) signaling pathways play important roles in interleukin (IL)-1β-induced matrix metalloproteinase (MMP)-9 activity and mRNA expression in house ear institute-organ of Corti 1 (HEI-OC1) cells. IL-1β induces MMP-9 activity and mRNA expression via the p38 MAPK signaling. (A) HEI-OC1 cells were pretreated with 50 µM ERK inhibitor (PD98059), 10 µM p38 MAPK inhibitor (SB203580), and 20 µM JNK inhibitor (SP600125), and then stimulated with 1 ng/mL IL-1β for 20 hours. Conditional media were collected and gelatin zymography was performed (A). The MMP-9 mRNA expression levels were determined by reverse transcription-polymerase chain reaction. The levels of actin were used as a loading control. The values in (B) represent the mean±SD from three independent samples. *P<0.001 compared to IL-1β. The data represent three independent experiments.

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