Inflammatory cytokines associated with degenerative disc disease control aggrecanase-1 (ADAMTS-4) expression in nucleus pulposus cells through MAPK and NF-κB

Ye Tian, Wen Yuan, Nobuyuki Fujita, Jianru Wang, Hua Wang, Irving M. Shapiro, Makarand V. Risbud

Research output: Contribution to journalArticle

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Abstract

We investigated TNF-α and IL-1β regulation of ADAMTS-4 expression in nucleus pulposus (NP) cells and its role in aggrecan degradation. Real-time quantitative RT-PCR, Western blotting, and transient transfections with rat NP cells and lentiviral silencing with human NP cells were performed to determine the roles of MAPK and NF-κB in cytokine-mediated ADAMTS-4 expression and function. ADAMTS4 expression and promoter activity increased in NP cells after TNF-α and IL-1β treatment. Treatment of cells with MAPK and NF-κB inhibitors abolished the inductive effect of the cytokines on ADAMTS4 mRNA and protein expression. Although ERK1, p38α, p38β2, and p38γ were involved in induction, ERK2 and p38δ played no role in TNF-α-dependent promoter activity. The inductive effect of p65 on ADAMTS4 promoter was confirmed through gain and loss-of-function studies. Cotransfection of p50 completely blocked p65-mediated induction. Lentiviral transduction with shRNA plasmids shp65, shp52, shIKK-α, and shIKK-β significantly decreased TNF-α-dependent increase in ADAMTS-4 and -5 levels and aggrecan degradation. Silencing of either ADAMTS-4 or -5 resulted in reduction in TNF-α-dependent aggrecan degradation in NP cells. By controlling activation of MAPK and NF-κB signaling, TNF-α and IL-1β modulate expression of ADAMTS-4 in NP cells. To our knowledge, this is the first study to show nonredundant contribution of both ADAMTS-4 and ADAMTS-5 to aggrecan degradation in human NP cells in vitro.

Original languageEnglish
Pages (from-to)2310-2321
Number of pages12
JournalAmerican Journal of Pathology
Volume182
Issue number6
DOIs
Publication statusPublished - 2013 Jun

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Cytokines
Aggrecans
Interleukin-1
ADAMTS4 Protein
Nucleus Pulposus
Small Interfering RNA
Transfection
Real-Time Polymerase Chain Reaction
Plasmids
Western Blotting
Messenger RNA

ASJC Scopus subject areas

  • Pathology and Forensic Medicine

Cite this

Inflammatory cytokines associated with degenerative disc disease control aggrecanase-1 (ADAMTS-4) expression in nucleus pulposus cells through MAPK and NF-κB. / Tian, Ye; Yuan, Wen; Fujita, Nobuyuki; Wang, Jianru; Wang, Hua; Shapiro, Irving M.; Risbud, Makarand V.

In: American Journal of Pathology, Vol. 182, No. 6, 06.2013, p. 2310-2321.

Research output: Contribution to journalArticle

Tian, Ye ; Yuan, Wen ; Fujita, Nobuyuki ; Wang, Jianru ; Wang, Hua ; Shapiro, Irving M. ; Risbud, Makarand V. / Inflammatory cytokines associated with degenerative disc disease control aggrecanase-1 (ADAMTS-4) expression in nucleus pulposus cells through MAPK and NF-κB. In: American Journal of Pathology. 2013 ; Vol. 182, No. 6. pp. 2310-2321.
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