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Interleukin-1 Receptor Antagonist Has a Novel Function in the Regulation of Matrix Metalloproteinase-13 Expression.

Goto H, Ishihara Y, Kikuchi T, Izawa A, Ozeki N, Okabe E, Kamiya Y, Ozawa Y, Mizutani H, Yamamoto G, Mogi M, Nakata K, Maeda H, Noguchi T, Mitani A - PLoS ONE (2015)

Bottom Line: These results suggested that continuous supply of IL-1 had no effect on the induction of MMP-13 by IL-1Ra siRNA.Laminin-5, which is degraded by MMP-13, was found in the internal basal lamina of wild-type mice, whereas the internal basal lamina of IL-1Ra KO mice did not show obvious laminin-5 localization.In conclusion, IL-1Ra is associated with MMP-13 expression and has a novel function in such regulation without interference of the IL-1 signaling cascade.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontology, School of Dentistry, Aichi Gakuin University, Chikusa-ku, Nagoya, Aichi, Japan.

ABSTRACT
Interleukin-1 receptor antagonist (IL-1Ra) is an IL-1 family member, which binds to IL-1 receptors but does not induce any intracellular signaling. We addressed whether IL-1Ra has a novel function in regulation of the extracellular matrix or adhesion molecules. Polymerase chain reaction array analysis demonstrated a ~5-fold increase in matrix metalloproteinase 13 (MMP-13) mRNA expression of IL-1Ra siRNA-transfected Ca9-22 human oral squamous epithelial carcinoma cells compared with the control. In fact, MMP-13 mRNA and protein expression as well as its activity in IL-1Ra siRNA-transfected Ca9-22 cell lines were significantly higher than those in the control. IL-1Ra siRNA treatment resulted in strong elevation of MMP-13 expression, whereas addition of rhIL-1Ra (40 ng/ml) suppressed MMP-13 expression, suggesting that IL-1Ra had a specific effect on MMP-13 induction. IL-1Ra siRNA could potently suppress IL-1α. No significant difference was found between the MMP-13 mRNA expression of IL-1Ra siRNA-transfected cells and those treated with anti-IL-1α or anti-IL-1β antibodies. These results suggested that continuous supply of IL-1 had no effect on the induction of MMP-13 by IL-1Ra siRNA. Histopathological investigation of MMP-13 in periodontal tissue showed specific localization in the junctional epithelial cells of IL-1Ra knockout (KO) mice. Furthermore, infection with Aggregatibacter actinomycetemcomitans to establish an experimental periodontitis model resulted in predominant localization of MMP-13 along apical junctional epithelial cells. Laminin-5, which is degraded by MMP-13, was found in the internal basal lamina of wild-type mice, whereas the internal basal lamina of IL-1Ra KO mice did not show obvious laminin-5 localization. In particular, laminin-5 localization almost disappeared in the internal basal lamina of IL-1Ra KO mice infected with A. actinomycetemcomitans, suggesting that the suppression of IL-1Ra resulted in strong induction of MMP-13 that degraded laminin-5. In conclusion, IL-1Ra is associated with MMP-13 expression and has a novel function in such regulation without interference of the IL-1 signaling cascade.

No MeSH data available.


Related in: MedlinePlus

Neutralization of IL-1 does not affect MMP-13 up-regulation.(A) IL-1α and (B) IL-1β mRNA (upper column) and protein expression (lower column) in IL-1Ra siRNA-transfected cells. The cells were cultured for 1, 3, 6, 12, and 24 hours, and then analyzed by real-time PCR. The protein levels in cells cultured for 24 hours were determined by an ELISA. Values represent fold changes (real-time PCR) or concentrations (ELISA). Differences among groups were analyzed using the Student’s t-test. Data are expressed as the mean ± SD (n = 3). *p < 0.05 vs each time point control. (C) MMP-13 mRNA expression in IL-1Ra and control siRNA-transfected cells treated with or without anti-IL-1α, anti-IL-1β, or isotype control antibodies (1 μg/ml). Cells were cultured for 6 hours, and then mRNA levels were determined using real-time PCR. Values represent fold changes. Differences among groups were analyzed by one-factor ANOVA and Bonferroni’s multiple comparison test. Data are expressed as the mean ± SD (n = 3). *p < 0.05.
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pone.0140942.g006: Neutralization of IL-1 does not affect MMP-13 up-regulation.(A) IL-1α and (B) IL-1β mRNA (upper column) and protein expression (lower column) in IL-1Ra siRNA-transfected cells. The cells were cultured for 1, 3, 6, 12, and 24 hours, and then analyzed by real-time PCR. The protein levels in cells cultured for 24 hours were determined by an ELISA. Values represent fold changes (real-time PCR) or concentrations (ELISA). Differences among groups were analyzed using the Student’s t-test. Data are expressed as the mean ± SD (n = 3). *p < 0.05 vs each time point control. (C) MMP-13 mRNA expression in IL-1Ra and control siRNA-transfected cells treated with or without anti-IL-1α, anti-IL-1β, or isotype control antibodies (1 μg/ml). Cells were cultured for 6 hours, and then mRNA levels were determined using real-time PCR. Values represent fold changes. Differences among groups were analyzed by one-factor ANOVA and Bonferroni’s multiple comparison test. Data are expressed as the mean ± SD (n = 3). *p < 0.05.

Mentions: IL-1α and IL-1β induce MMP-13 production via the IL-1 receptor [23, 24]. We found continuous production of IL-1α (35 pg/mL) and IL-1β (4 pg/mL) in Ca9-22 cells (Fig 6A and 6B). By examining whether IL-1Ra can regulate IL-1 production in these cells, we found that IL-1Ra siRNA resulted in a strong but transient decrease in only IL-1α production (Fig 6A), indicating that IL-1Ra regulates the production of IL-1α. It is likely that these proinflammatory cytokines had no effect on MMP-13 production under similar conditions.


Interleukin-1 Receptor Antagonist Has a Novel Function in the Regulation of Matrix Metalloproteinase-13 Expression.

Goto H, Ishihara Y, Kikuchi T, Izawa A, Ozeki N, Okabe E, Kamiya Y, Ozawa Y, Mizutani H, Yamamoto G, Mogi M, Nakata K, Maeda H, Noguchi T, Mitani A - PLoS ONE (2015)

Neutralization of IL-1 does not affect MMP-13 up-regulation.(A) IL-1α and (B) IL-1β mRNA (upper column) and protein expression (lower column) in IL-1Ra siRNA-transfected cells. The cells were cultured for 1, 3, 6, 12, and 24 hours, and then analyzed by real-time PCR. The protein levels in cells cultured for 24 hours were determined by an ELISA. Values represent fold changes (real-time PCR) or concentrations (ELISA). Differences among groups were analyzed using the Student’s t-test. Data are expressed as the mean ± SD (n = 3). *p < 0.05 vs each time point control. (C) MMP-13 mRNA expression in IL-1Ra and control siRNA-transfected cells treated with or without anti-IL-1α, anti-IL-1β, or isotype control antibodies (1 μg/ml). Cells were cultured for 6 hours, and then mRNA levels were determined using real-time PCR. Values represent fold changes. Differences among groups were analyzed by one-factor ANOVA and Bonferroni’s multiple comparison test. Data are expressed as the mean ± SD (n = 3). *p < 0.05.
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Related In: Results  -  Collection

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pone.0140942.g006: Neutralization of IL-1 does not affect MMP-13 up-regulation.(A) IL-1α and (B) IL-1β mRNA (upper column) and protein expression (lower column) in IL-1Ra siRNA-transfected cells. The cells were cultured for 1, 3, 6, 12, and 24 hours, and then analyzed by real-time PCR. The protein levels in cells cultured for 24 hours were determined by an ELISA. Values represent fold changes (real-time PCR) or concentrations (ELISA). Differences among groups were analyzed using the Student’s t-test. Data are expressed as the mean ± SD (n = 3). *p < 0.05 vs each time point control. (C) MMP-13 mRNA expression in IL-1Ra and control siRNA-transfected cells treated with or without anti-IL-1α, anti-IL-1β, or isotype control antibodies (1 μg/ml). Cells were cultured for 6 hours, and then mRNA levels were determined using real-time PCR. Values represent fold changes. Differences among groups were analyzed by one-factor ANOVA and Bonferroni’s multiple comparison test. Data are expressed as the mean ± SD (n = 3). *p < 0.05.
Mentions: IL-1α and IL-1β induce MMP-13 production via the IL-1 receptor [23, 24]. We found continuous production of IL-1α (35 pg/mL) and IL-1β (4 pg/mL) in Ca9-22 cells (Fig 6A and 6B). By examining whether IL-1Ra can regulate IL-1 production in these cells, we found that IL-1Ra siRNA resulted in a strong but transient decrease in only IL-1α production (Fig 6A), indicating that IL-1Ra regulates the production of IL-1α. It is likely that these proinflammatory cytokines had no effect on MMP-13 production under similar conditions.

Bottom Line: These results suggested that continuous supply of IL-1 had no effect on the induction of MMP-13 by IL-1Ra siRNA.Laminin-5, which is degraded by MMP-13, was found in the internal basal lamina of wild-type mice, whereas the internal basal lamina of IL-1Ra KO mice did not show obvious laminin-5 localization.In conclusion, IL-1Ra is associated with MMP-13 expression and has a novel function in such regulation without interference of the IL-1 signaling cascade.

View Article: PubMed Central - PubMed

Affiliation: Department of Periodontology, School of Dentistry, Aichi Gakuin University, Chikusa-ku, Nagoya, Aichi, Japan.

ABSTRACT
Interleukin-1 receptor antagonist (IL-1Ra) is an IL-1 family member, which binds to IL-1 receptors but does not induce any intracellular signaling. We addressed whether IL-1Ra has a novel function in regulation of the extracellular matrix or adhesion molecules. Polymerase chain reaction array analysis demonstrated a ~5-fold increase in matrix metalloproteinase 13 (MMP-13) mRNA expression of IL-1Ra siRNA-transfected Ca9-22 human oral squamous epithelial carcinoma cells compared with the control. In fact, MMP-13 mRNA and protein expression as well as its activity in IL-1Ra siRNA-transfected Ca9-22 cell lines were significantly higher than those in the control. IL-1Ra siRNA treatment resulted in strong elevation of MMP-13 expression, whereas addition of rhIL-1Ra (40 ng/ml) suppressed MMP-13 expression, suggesting that IL-1Ra had a specific effect on MMP-13 induction. IL-1Ra siRNA could potently suppress IL-1α. No significant difference was found between the MMP-13 mRNA expression of IL-1Ra siRNA-transfected cells and those treated with anti-IL-1α or anti-IL-1β antibodies. These results suggested that continuous supply of IL-1 had no effect on the induction of MMP-13 by IL-1Ra siRNA. Histopathological investigation of MMP-13 in periodontal tissue showed specific localization in the junctional epithelial cells of IL-1Ra knockout (KO) mice. Furthermore, infection with Aggregatibacter actinomycetemcomitans to establish an experimental periodontitis model resulted in predominant localization of MMP-13 along apical junctional epithelial cells. Laminin-5, which is degraded by MMP-13, was found in the internal basal lamina of wild-type mice, whereas the internal basal lamina of IL-1Ra KO mice did not show obvious laminin-5 localization. In particular, laminin-5 localization almost disappeared in the internal basal lamina of IL-1Ra KO mice infected with A. actinomycetemcomitans, suggesting that the suppression of IL-1Ra resulted in strong induction of MMP-13 that degraded laminin-5. In conclusion, IL-1Ra is associated with MMP-13 expression and has a novel function in such regulation without interference of the IL-1 signaling cascade.

No MeSH data available.


Related in: MedlinePlus