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Expression of Proteinase-Activated Receptor-2 in the Intervertebral Disc

Iida, Ryu MD*; Akeda, Koji MD, PhD*; Kasai, Yuichi MD, PhD*; Masuda, Koichi MD; Morimoto, Ryo MD*; Sakakibara, Toshihiko MD; Sato, Masayoshi MD, PhD; Uchida, Atsumasa MD, PhD*

doi: 10.1097/BRS.0b013e318195a67d
Basic Science

Study Design. Immunohistochemical and biochemical analyses of proteinase-activated receptor-2 (PAR-2) in rat and human intervertebral discs (IVDs).

Objectives. To examine the expression and function of PAR-2 in rat IVD cells, and to determine if PAR-2 is expressed in human IVDs.

Summary of Background Data. PAR-2 is a G protein-coupled receptor that contributes to the regulation of inflammatory reactions and the pathophysiology of inflammatory diseases, including arthritis. The expression of PAR-2 in the IVD has not been determined.

Methods. PAR-2 expression by rat IVD cells and tissues was examined using immunohistochemistry and western blot. Rat anulus fibrosus cells in monolayer culture were used to examine the biologic role of PAR-2 in vitro. The effect of PAR-2–activating peptide (PAR-2AP) on the catabolic cascade was assessed by western blot and real-time PCR. The expression of PAR-2 by human IVD tissues at different stages of degeneration was determined by immunohistochemical analyses.

Results. PAR-2 was expressed by rat IVD cells and in both anulus fibrosus and nucleus pulposus tissues, PAR-2 expression was up-regulated by interleukin-1β (IL-1β). PAR-2AP significantly increased the release of IL-1β into the medium. Although PAR-2AP had no direct effect on matrix metalloproteinase-3 (MMP-3) and MMP-13 mRNA levels, treatment with PAR-2AP significantly up-regulated the mRNA levels of a disintegrin and metalloproteinase with thrombospondin motif–4. The simultaneous administration of PAR-2AP and IL-1β synergistically up-regulated the mRNA levels of a disintegrin and metalloproteinase with thrombospondin motif–4, MMP-3, and MMP-13. The expression of PAR-2 was identified in human IVD tissues. The number of PAR-2–expressing cells was significantly elevated in advanced stages of IVD degeneration compared with those in early stages of degeneration.

Conclusion. Our results demonstrate for the first time that IVD cells express PAR-2. The expression of PAR-2 is regulated by IL-1β stimulation. PAR-2 activation accelerates the expression of matrix-degrading enzymes. PAR-2 may play an important role in the cytokine-mediated catabolic cascade and consequently may be involved in IVD degeneration.

Proteinase-activated receptor-2 (PAR-2) expression was identified in human and rat intervertebral discs (IVDs). PAR-2 activation accelerated the catabolic pathway in cultured rat IVD cells. The expression of PAR-2 in human IVD tissues was elevated in tissues with advanced stages of degeneration compared with tissues with early stages of degeneration.

From the *Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, Mie, Japan; †Department of Orthopedic Surgery, University of California, San Diego CA; and ‡Department of Orthopaedic Surgery, Murase Hospital, Suzuka, Japan.

Acknowledgment date: August 12, 2008. Revision date: September 27, 2008. Acceptance date: September 28, 2008.

The manuscript submitted does not contain information about medical device(s)/drug(s).

No funds were received in support of this work. No benefits in any form have been or will be received from a commercial party related directly or indirectly to the subject of this manuscript.

This study was sponsored by grants from the Ministry of Education, Culture, Sports, and Science and Technology (Japan).

Address correspondence and reprint requests to Koji Akeda, MD, PhD, Department of Orthopaedic Surgery, Mie University Graduate School of Medicine, 2–174 Edobashi, Tsu city, Mie, Japan; E-mail:

© 2009 Lippincott Williams & Wilkins, Inc.