Substance P Receptor Antagonist Suppresses Inflammatory Cytokine Expression in Human Disc Cells.

Authors

• Christopher K Kepler
• Dessislava Z Markova
• John D Koerner
• Joseph Mendelis
• Chiu-Ming Chen
• Alexander R Vaccaro
• Makarand V Risbud
• Todd J Albert
• D Greg Anderson

Publication/Presentation Date

8-15-2015

Abstract

STUDY DESIGN: Laboratory study.

OBJECTIVE: To evaluate whether blockade of the Substance P (SP) NK1R attenuates its proinflammatory effect on human intervertebral disc cells (IVD), and to evaluate the signaling pathways associated with SP.

SUMMARY OF BACKGROUND DATA: SP and its receptors are expressed in human IVD cells, and cause upregulation of inflammatory mediators; however, the effects of blocking these receptors have not been studied in human IVD cells.

METHODS: Human annulus fibrosus (AF) and nucleus pulposus (NP) cells were expanded in monolayer, and then suspended in alginate beads. The alginate beads were treated with culture medium first containing a high affinity NK1R antagonist (L-760735) at different concentrations, and then with medium containing both NK1R antagonist and SP at 2 concentrations. Ribonucleic acid was isolated and transcribed into cDNA. Quantitative reverse transcriptase-polymerase chain reaction (RT-PCR) was performed to evaluate expression of interleukin (IL)-1β, IL-6, and IL-8. Western blot analysis was performed to examine levels of the phosphorylated p38 mitogen-activated protein kinase (MAPK), extracellular signal regulated kinase 1/2 (ERK1/2) and nuclear factor kappa-light-chain-enhancer of activated B cells (NFκB p65). The cells were pretreated with specific inhibitors of p38 (SB203580), ERK1/2 (PD98059), and p65 (SM7368) and then stimulated with SP.

RESULTS: We detected expression of NK1R, neurokinin receptor 2 (NK2R), and neurokinin receptor 3 (NK3R) in AF and NP cells. Treatment of disc cells with the NK1R antagonist was able to suppress expression of IL-1β, IL-6, and IL-8 in a dose-dependent manner. SP stimulation increased phosphorylation of p38-MAPK and ERK1/2, but not of NFκB p65. This indicates that p38-MAPK and ERK1/2 control SP-induced cytokine expression independently from NF-kB p65. Inhibition of p38 and ERK1/2 activation reduced SP-induced IL-6 production in human disc cells.

CONCLUSION: NK1R is responsible for the proinflammatory effect of SP on IVD cells and this effect can be blocked by preventing binding of SP to NK1R. This study shows for the first time that SP mediates signaling in disc cells through NK1R and that SP activates the proinflammatory p38-MAPK and ERK1/2 pathways.

LEVEL OF EVIDENCE: 4.

Volume

40

Issue

16

First Page

1261

Last Page

1269

ISSN

1528-1159

Published In/Presented At

Kepler, C. K., Markova, D. Z., Koerner, J. D., Mendelis, J., Chen, C. M., Vaccaro, A. R., Risbud, M. V., Albert, T. J., & Anderson, D. G. (2015). Substance P Receptor Antagonist Suppresses Inflammatory Cytokine Expression in Human Disc Cells. Spine, 40(16), 1261–1269. https://doi.org/10.1097/BRS.0000000000000954

Disciplines

Business Administration, Management, and Operations | Health and Medical Administration | Management Sciences and Quantitative Methods

PubMedID

25929203

Department(s)

Administration and Leadership

Document Type

Article