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IL-1β promotes ADAMTS enzyme-mediated aggrecan degradation through NF-κB in human intervertebral disc.

Sun Z, Yin Z, Liu C, Liang H, Jiang M, Tian J - J Orthop Surg Res (2015)

Bottom Line: Treatment with NF-κB inhibitors abolished the inductive effect of the cytokines on ADAMTS expression.Silencing of p65 confirmed their role in IL-1β-dependent ADAMTS-4 and ADAMTS-5 expression and aggrecan degradation.To our knowledge, this is the first study that shows the contribution of both ADAMTS-4 and ADAMTS-5 to aggrecan degradation in human NP cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, School of Medicine, Shanghai General Hospital Affiliated to Shanghai Jiao Tong University, 100, Haining Road, Shanghai, 200080, China. sunzhy0008@163.com.

ABSTRACT

Background: The purpose of this study is to investigate IL-1β regulation of a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS-4 and ADAMTS-5) expression through nuclear factor kappa B (NF-κB) in human nucleus pulposus (NP) cells.

Methods: qRT-PCR and Western blot were used to measure ADAMTS expression. Transfections and gene silencing were used to determine the role of NF-κB on cytokine-mediated ADAMTS expression and its role in aggrecan degradation.

Results: IL-1β increased ADAMTS expression in NP cells. Treatment with NF-κB inhibitors abolished the inductive effect of the cytokines on ADAMTS expression. Silencing of p65 confirmed their role in IL-1β-dependent ADAMTS-4 and ADAMTS-5 expression and aggrecan degradation.

Conclusions: By controlling the activation of NF-κB signaling, IL-1β modulates the expression of ADAMTS in NP cells. To our knowledge, this is the first study that shows the contribution of both ADAMTS-4 and ADAMTS-5 to aggrecan degradation in human NP cells.

No MeSH data available.


Modulation of IL-1β-dependent expression of ADAMTS-4 and ADAMTS-5 expression by NF-κB signaling in human NP cells. a Western blot analysis of p65 nucleoproteins after treatment of NP cells with IL-1β. b Western blot analysis indicates that treatment with NF-κB inhibitor completely abolished ADAMTS-4 and ADAMTS-5 protein induction by IL-1β and the level of aggrecan was significantly increased. Data are expressed as mean ± SD from three independent experiments
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Fig5: Modulation of IL-1β-dependent expression of ADAMTS-4 and ADAMTS-5 expression by NF-κB signaling in human NP cells. a Western blot analysis of p65 nucleoproteins after treatment of NP cells with IL-1β. b Western blot analysis indicates that treatment with NF-κB inhibitor completely abolished ADAMTS-4 and ADAMTS-5 protein induction by IL-1β and the level of aggrecan was significantly increased. Data are expressed as mean ± SD from three independent experiments

Mentions: To determine whether NF-κB signaling is required for the cytokine-dependent induction of ADAMTS-4 and ADAMTS-5 in human NP cells, we first evaluated the activation of NF-κB signaling pathways after treatment with IL-1β. After treatment with IL-1β, there was a rapid increase in p65 nucleoprotein levels (Fig. 5a). To ascertain whether the IL-1β-induced expression of ADAMTS-4 and ADAMTS-5 requires NF-κB signaling, human NP cells were pretreated with pathway-specific inhibitors. Pretreatment caused a significant suppression in the IL-1β induction of both ADAMTS-4 and ADAMTS-5 mRNA levels (Fig. 6a, b). Similarly, a pronounced decrease in the IL-1β-mediated increase in the levels of the ADAMTS-4 and ADAMTS-5 protein was seen in the presence of NF-κB pathway inhibitors (Fig. 5b). Importantly, we examined the effect of ADAMTS-4 and ADAMTS-5 expression on aggrecan degradation in human NP cells. Suppression of ADAMTS-4 and ADAMTS-5 expression resulted in a significant inhibition of IL-1β-mediated aggrecan degradation in human NP cells (Figs. 5b and 6c).Fig. 5


IL-1β promotes ADAMTS enzyme-mediated aggrecan degradation through NF-κB in human intervertebral disc.

Sun Z, Yin Z, Liu C, Liang H, Jiang M, Tian J - J Orthop Surg Res (2015)

Modulation of IL-1β-dependent expression of ADAMTS-4 and ADAMTS-5 expression by NF-κB signaling in human NP cells. a Western blot analysis of p65 nucleoproteins after treatment of NP cells with IL-1β. b Western blot analysis indicates that treatment with NF-κB inhibitor completely abolished ADAMTS-4 and ADAMTS-5 protein induction by IL-1β and the level of aggrecan was significantly increased. Data are expressed as mean ± SD from three independent experiments
© Copyright Policy - OpenAccess
Related In: Results  -  Collection

License 1 - License 2
Show All Figures
getmorefigures.php?uid=PMC4594913&req=5

Fig5: Modulation of IL-1β-dependent expression of ADAMTS-4 and ADAMTS-5 expression by NF-κB signaling in human NP cells. a Western blot analysis of p65 nucleoproteins after treatment of NP cells with IL-1β. b Western blot analysis indicates that treatment with NF-κB inhibitor completely abolished ADAMTS-4 and ADAMTS-5 protein induction by IL-1β and the level of aggrecan was significantly increased. Data are expressed as mean ± SD from three independent experiments
Mentions: To determine whether NF-κB signaling is required for the cytokine-dependent induction of ADAMTS-4 and ADAMTS-5 in human NP cells, we first evaluated the activation of NF-κB signaling pathways after treatment with IL-1β. After treatment with IL-1β, there was a rapid increase in p65 nucleoprotein levels (Fig. 5a). To ascertain whether the IL-1β-induced expression of ADAMTS-4 and ADAMTS-5 requires NF-κB signaling, human NP cells were pretreated with pathway-specific inhibitors. Pretreatment caused a significant suppression in the IL-1β induction of both ADAMTS-4 and ADAMTS-5 mRNA levels (Fig. 6a, b). Similarly, a pronounced decrease in the IL-1β-mediated increase in the levels of the ADAMTS-4 and ADAMTS-5 protein was seen in the presence of NF-κB pathway inhibitors (Fig. 5b). Importantly, we examined the effect of ADAMTS-4 and ADAMTS-5 expression on aggrecan degradation in human NP cells. Suppression of ADAMTS-4 and ADAMTS-5 expression resulted in a significant inhibition of IL-1β-mediated aggrecan degradation in human NP cells (Figs. 5b and 6c).Fig. 5

Bottom Line: Treatment with NF-κB inhibitors abolished the inductive effect of the cytokines on ADAMTS expression.Silencing of p65 confirmed their role in IL-1β-dependent ADAMTS-4 and ADAMTS-5 expression and aggrecan degradation.To our knowledge, this is the first study that shows the contribution of both ADAMTS-4 and ADAMTS-5 to aggrecan degradation in human NP cells.

View Article: PubMed Central - PubMed

Affiliation: Department of Orthopedics, School of Medicine, Shanghai General Hospital Affiliated to Shanghai Jiao Tong University, 100, Haining Road, Shanghai, 200080, China. sunzhy0008@163.com.

ABSTRACT

Background: The purpose of this study is to investigate IL-1β regulation of a disintegrin and metalloproteinase with thrombospondin motifs (ADAMTS-4 and ADAMTS-5) expression through nuclear factor kappa B (NF-κB) in human nucleus pulposus (NP) cells.

Methods: qRT-PCR and Western blot were used to measure ADAMTS expression. Transfections and gene silencing were used to determine the role of NF-κB on cytokine-mediated ADAMTS expression and its role in aggrecan degradation.

Results: IL-1β increased ADAMTS expression in NP cells. Treatment with NF-κB inhibitors abolished the inductive effect of the cytokines on ADAMTS expression. Silencing of p65 confirmed their role in IL-1β-dependent ADAMTS-4 and ADAMTS-5 expression and aggrecan degradation.

Conclusions: By controlling the activation of NF-κB signaling, IL-1β modulates the expression of ADAMTS in NP cells. To our knowledge, this is the first study that shows the contribution of both ADAMTS-4 and ADAMTS-5 to aggrecan degradation in human NP cells.

No MeSH data available.