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SERPINE2 Inhibits IL-1α-Induced MMP-13 Expression in Human Chondrocytes: Involvement of ERK/NF-κB/AP-1 Pathways.

Santoro A, Conde J, Scotece M, Abella V, Lois A, Lopez V, Pino J, Gomez R, Gomez-Reino JJ, Gualillo O - PLoS ONE (2015)

Bottom Line: We also examined the effects of SERPINE2 on IL-1α-induced MMP-13 expression.This inhibitory effect is likely regulated through a pathway involving ERK 1/2, NF-κB and AP-1.Taken together, these data demonstrate that SERPINE2 might prevent cartilage catabolism by inhibiting the expression of MMP-13, one of the most relevant collagenases, involved in cartilage breakdown in OA.

View Article: PubMed Central - PubMed

Affiliation: SERGAS (Servizo Galego de Saude) and IDIS (Instituto de Investigación Sanitaria de Santiago), the NEIRID Lab (Neuroendocrine Interactions in Rheumatology and Inflammatory Diseases), Research Laboratory 9, Santiago University Clinical Hospital, Santiago de Compostela, Spain; University of Naples Federico II, Dept. of Pharmacy, 80138, Naples, Italy.

ABSTRACT

Objectives: Osteoarthritis (OA) is a chronic joint disease, characterized by a progressive loss of articular cartilage. During OA, proinflammatory cytokines, such as interleukin IL-1, induce the expression of matrix metalloproteinases (MMPs) in chondrocytes, contributing thus to the extracellular matrix (ECM) degradation. Members of Serpine family, including plasminogen activator inhibitors have been reported to participate in ECM regulation. The aim of this study was to assess the expression of serpin peptidase inhibitor clade E member 2 (SERPINE2), under basal conditions and in response to increasing doses of IL-1α, in human cultured chondrocytes. We also examined the effects of SERPINE2 on IL-1α-induced MMP-13 expression. For completeness, the signaling pathway involved in this process was also explored.

Methods: SERPINE2 mRNA and protein expression were evaluated by RT-qPCR and western blot analysis in human T/C-28a2 cell line and human primary chondrocytes. These cells were treated with human recombinant SERPINE2, alone or in combination with IL-1α. ERK 1/2, NFκB and AP-1 activation were assessed by western blot analysis.

Results: Human cultured chondrocytes express SERPINE2 in basal condition. This expression increased in response to IL-1α stimulation. In addition, recombinant SERPINE2 induced a clear inhibition of MMP-13 expression in IL-1α-stimulated chondrocytes. This inhibitory effect is likely regulated through a pathway involving ERK 1/2, NF-κB and AP-1.

Conclusions: Taken together, these data demonstrate that SERPINE2 might prevent cartilage catabolism by inhibiting the expression of MMP-13, one of the most relevant collagenases, involved in cartilage breakdown in OA.

No MeSH data available.


Related in: MedlinePlus

A. Human MMP-13 mRNA expression in T/C-28a2 chondrocytes transfected with a siRNA against SERPINE2 (siSERPINE2, 10nM) or a non-targeting control siRNA (siC-, 10 nM) in presence or not of IL-1α (0.5 ng/mL) for 24 h. B. MMP-13 endogenous catalytic activity was measured in the conditioned medium of OA human primary chondrocytes after recombinant SERPINE2 (0.4 ng/mL) treatment in presence or not of IL-1α (0.5 ng/mL) for 24 h.*P<0.05 and ***P<0.001 vs siC- transfected cells; ### P<0.001 vs siC- plus IL-1α-stimulated chondrocytes.
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pone.0135979.g003: A. Human MMP-13 mRNA expression in T/C-28a2 chondrocytes transfected with a siRNA against SERPINE2 (siSERPINE2, 10nM) or a non-targeting control siRNA (siC-, 10 nM) in presence or not of IL-1α (0.5 ng/mL) for 24 h. B. MMP-13 endogenous catalytic activity was measured in the conditioned medium of OA human primary chondrocytes after recombinant SERPINE2 (0.4 ng/mL) treatment in presence or not of IL-1α (0.5 ng/mL) for 24 h.*P<0.05 and ***P<0.001 vs siC- transfected cells; ### P<0.001 vs siC- plus IL-1α-stimulated chondrocytes.

Mentions: To address the role of endogenous SERPINE2 on MMP-13 expression we used a different approach. In this case, we silenced the expression of SERPINE2 by using a siRNA against this serine protease. As shown in Fig 3A, SERPINE2 gene knockdown led to an increase in MMP-13 expression in comparison to cells transfected with a non-targeting control siRNA. We also observed that SERPINE2 silencing produced a strong up-regulation of MMP-13 mRNA expression in IL-1α treated chondrocytes (Fig 3A).


SERPINE2 Inhibits IL-1α-Induced MMP-13 Expression in Human Chondrocytes: Involvement of ERK/NF-κB/AP-1 Pathways.

Santoro A, Conde J, Scotece M, Abella V, Lois A, Lopez V, Pino J, Gomez R, Gomez-Reino JJ, Gualillo O - PLoS ONE (2015)

A. Human MMP-13 mRNA expression in T/C-28a2 chondrocytes transfected with a siRNA against SERPINE2 (siSERPINE2, 10nM) or a non-targeting control siRNA (siC-, 10 nM) in presence or not of IL-1α (0.5 ng/mL) for 24 h. B. MMP-13 endogenous catalytic activity was measured in the conditioned medium of OA human primary chondrocytes after recombinant SERPINE2 (0.4 ng/mL) treatment in presence or not of IL-1α (0.5 ng/mL) for 24 h.*P<0.05 and ***P<0.001 vs siC- transfected cells; ### P<0.001 vs siC- plus IL-1α-stimulated chondrocytes.
© Copyright Policy
Related In: Results  -  Collection

License
Show All Figures
getmorefigures.php?uid=PMC4549255&req=5

pone.0135979.g003: A. Human MMP-13 mRNA expression in T/C-28a2 chondrocytes transfected with a siRNA against SERPINE2 (siSERPINE2, 10nM) or a non-targeting control siRNA (siC-, 10 nM) in presence or not of IL-1α (0.5 ng/mL) for 24 h. B. MMP-13 endogenous catalytic activity was measured in the conditioned medium of OA human primary chondrocytes after recombinant SERPINE2 (0.4 ng/mL) treatment in presence or not of IL-1α (0.5 ng/mL) for 24 h.*P<0.05 and ***P<0.001 vs siC- transfected cells; ### P<0.001 vs siC- plus IL-1α-stimulated chondrocytes.
Mentions: To address the role of endogenous SERPINE2 on MMP-13 expression we used a different approach. In this case, we silenced the expression of SERPINE2 by using a siRNA against this serine protease. As shown in Fig 3A, SERPINE2 gene knockdown led to an increase in MMP-13 expression in comparison to cells transfected with a non-targeting control siRNA. We also observed that SERPINE2 silencing produced a strong up-regulation of MMP-13 mRNA expression in IL-1α treated chondrocytes (Fig 3A).

Bottom Line: We also examined the effects of SERPINE2 on IL-1α-induced MMP-13 expression.This inhibitory effect is likely regulated through a pathway involving ERK 1/2, NF-κB and AP-1.Taken together, these data demonstrate that SERPINE2 might prevent cartilage catabolism by inhibiting the expression of MMP-13, one of the most relevant collagenases, involved in cartilage breakdown in OA.

View Article: PubMed Central - PubMed

Affiliation: SERGAS (Servizo Galego de Saude) and IDIS (Instituto de Investigación Sanitaria de Santiago), the NEIRID Lab (Neuroendocrine Interactions in Rheumatology and Inflammatory Diseases), Research Laboratory 9, Santiago University Clinical Hospital, Santiago de Compostela, Spain; University of Naples Federico II, Dept. of Pharmacy, 80138, Naples, Italy.

ABSTRACT

Objectives: Osteoarthritis (OA) is a chronic joint disease, characterized by a progressive loss of articular cartilage. During OA, proinflammatory cytokines, such as interleukin IL-1, induce the expression of matrix metalloproteinases (MMPs) in chondrocytes, contributing thus to the extracellular matrix (ECM) degradation. Members of Serpine family, including plasminogen activator inhibitors have been reported to participate in ECM regulation. The aim of this study was to assess the expression of serpin peptidase inhibitor clade E member 2 (SERPINE2), under basal conditions and in response to increasing doses of IL-1α, in human cultured chondrocytes. We also examined the effects of SERPINE2 on IL-1α-induced MMP-13 expression. For completeness, the signaling pathway involved in this process was also explored.

Methods: SERPINE2 mRNA and protein expression were evaluated by RT-qPCR and western blot analysis in human T/C-28a2 cell line and human primary chondrocytes. These cells were treated with human recombinant SERPINE2, alone or in combination with IL-1α. ERK 1/2, NFκB and AP-1 activation were assessed by western blot analysis.

Results: Human cultured chondrocytes express SERPINE2 in basal condition. This expression increased in response to IL-1α stimulation. In addition, recombinant SERPINE2 induced a clear inhibition of MMP-13 expression in IL-1α-stimulated chondrocytes. This inhibitory effect is likely regulated through a pathway involving ERK 1/2, NF-κB and AP-1.

Conclusions: Taken together, these data demonstrate that SERPINE2 might prevent cartilage catabolism by inhibiting the expression of MMP-13, one of the most relevant collagenases, involved in cartilage breakdown in OA.

No MeSH data available.


Related in: MedlinePlus