Induction of proinflammatory cytokine production in intervertebral disc cells by macrophage-like THP-1 cells requires mitogen-activated protein kinase activity

Jung Jae Park, Hong Joo Moon, Jin Hyun Park, Taek-Hyun Kwon, Youn-Kwan Park, Joo-Han Kim

Research output: Contribution to journalArticle

7 Citations (Scopus)

Abstract

OBJECTIVE: To determine the role played by mitogen-activated protein kinase (MAPK) signaling in the interactions between macrophages and intervertebral disc (IVD) cells, it was hypothesized that MAPK inhibition would modulate the production of the proinflammatory cytokines associated with inflammatory reaction in IVD cells. METHODS: Human annulus fibrosus (AF) and nucleus pulposus (NP) cells were cocultured with phorbol myristate acetate-stimulated macrophage-like THP-1 cells, with and without SB202190 (a p38-α and -β inhibitor), SP600125 (a c-Jun N-terminal kinase [JNK] inhibitor), and PD98059 (an extracellular signal-regulated kinase [ERK] 1/2 inhibitor). The cytokines in conditioned media from cocultured and macrophage-exposed (nemotic) cells were assayed using enzyme-linked immunosorbent assays (ELISAs). RESULTS: Interleukin (IL)-6 and IL-8 were secreted in greater quantities by the cocultured cells compared with naive IVD cells and macrophages (Mφ) cultured alone. The tumor necrosis factor (TNF)-α and IL-6 levels produced by the NP cells cocultured with Mφs (NP-Mφ) were significantly lower than those produced by AF cells cocultured with Mφs (AF-Mφ). SB202190 dose-dependently suppressed IL-6 secretion by AF-Mφ and NP-Mφ cocultures, and 10 μM SB202190 significantly decreased IL-6 and IL-8 production in nemotic AF and NP pellets. SP600125 at 10 μM significantly suppressed the production of TNFα, IL-6, and IL-8 in AF-Mφ and NP-Mφ cocultures and significantly suppressed IL-1β production in the NP-Mφ coculture. Administration of 10 μM PD98059 significantly decreased IL-6 levels in the AF-Mφ coculture, and decreased the levels of TNFα and IL-8 in both the AF-Mφ and NP-Mφ cocultures. CONCLUSIONS: The present study shows that inhibitors of p38 MAPK effectively controlled IL-6 production during inflammatory reactions and that JNK and ERK1/2 inhibitors successfully suppressed the production of major proinflammatory cytokines during interactions between macrophages and IVD cells. Therefore, selective blockade of these signals may serve as a therapeutic approach to symptomatic IVD degeneration.

Original languageEnglish
Pages (from-to)167-175
Number of pages9
JournalJournal of Neurosurgery: Spine
Volume24
Issue number1
DOIs
Publication statusPublished - 2016 Jan 1

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Intervertebral Disc
Mitogen-Activated Protein Kinases
Macrophages
Cytokines
Interleukin-6
Coculture Techniques
Interleukin-8
Mitogen-Activated Protein Kinase 3
Tumor Necrosis Factor-alpha
Intervertebral Disc Degeneration
Annulus Fibrosus
Nucleus Pulposus
JNK Mitogen-Activated Protein Kinases
Mitogen-Activated Protein Kinase 1
Tetradecanoylphorbol Acetate
p38 Mitogen-Activated Protein Kinases
Conditioned Culture Medium
Interleukin-1
Enzyme-Linked Immunosorbent Assay

Keywords

  • Cytokines
  • Intervertebral disc degeneration
  • Mitogen-activated protein kinases
  • PD98059
  • SB202190
  • SP600125

ASJC Scopus subject areas

  • Surgery
  • Neurology
  • Clinical Neurology

Cite this

@article{0da1708a38274d14b734a68e0d1a07f1,
title = "Induction of proinflammatory cytokine production in intervertebral disc cells by macrophage-like THP-1 cells requires mitogen-activated protein kinase activity",
abstract = "OBJECTIVE: To determine the role played by mitogen-activated protein kinase (MAPK) signaling in the interactions between macrophages and intervertebral disc (IVD) cells, it was hypothesized that MAPK inhibition would modulate the production of the proinflammatory cytokines associated with inflammatory reaction in IVD cells. METHODS: Human annulus fibrosus (AF) and nucleus pulposus (NP) cells were cocultured with phorbol myristate acetate-stimulated macrophage-like THP-1 cells, with and without SB202190 (a p38-α and -β inhibitor), SP600125 (a c-Jun N-terminal kinase [JNK] inhibitor), and PD98059 (an extracellular signal-regulated kinase [ERK] 1/2 inhibitor). The cytokines in conditioned media from cocultured and macrophage-exposed (nemotic) cells were assayed using enzyme-linked immunosorbent assays (ELISAs). RESULTS: Interleukin (IL)-6 and IL-8 were secreted in greater quantities by the cocultured cells compared with naive IVD cells and macrophages (Mφ) cultured alone. The tumor necrosis factor (TNF)-α and IL-6 levels produced by the NP cells cocultured with Mφs (NP-Mφ) were significantly lower than those produced by AF cells cocultured with Mφs (AF-Mφ). SB202190 dose-dependently suppressed IL-6 secretion by AF-Mφ and NP-Mφ cocultures, and 10 μM SB202190 significantly decreased IL-6 and IL-8 production in nemotic AF and NP pellets. SP600125 at 10 μM significantly suppressed the production of TNFα, IL-6, and IL-8 in AF-Mφ and NP-Mφ cocultures and significantly suppressed IL-1β production in the NP-Mφ coculture. Administration of 10 μM PD98059 significantly decreased IL-6 levels in the AF-Mφ coculture, and decreased the levels of TNFα and IL-8 in both the AF-Mφ and NP-Mφ cocultures. CONCLUSIONS: The present study shows that inhibitors of p38 MAPK effectively controlled IL-6 production during inflammatory reactions and that JNK and ERK1/2 inhibitors successfully suppressed the production of major proinflammatory cytokines during interactions between macrophages and IVD cells. Therefore, selective blockade of these signals may serve as a therapeutic approach to symptomatic IVD degeneration.",
keywords = "Cytokines, Intervertebral disc degeneration, Mitogen-activated protein kinases, PD98059, SB202190, SP600125",
author = "Park, {Jung Jae} and Moon, {Hong Joo} and Park, {Jin Hyun} and Taek-Hyun Kwon and Youn-Kwan Park and Joo-Han Kim",
year = "2016",
month = "1",
day = "1",
doi = "10.3171/2015.3.SPINE14729",
language = "English",
volume = "24",
pages = "167--175",
journal = "Journal of Neurosurgery: Spine",
issn = "1547-5654",
publisher = "American Association of Neurological Surgeons",
number = "1",

}

TY - JOUR

T1 - Induction of proinflammatory cytokine production in intervertebral disc cells by macrophage-like THP-1 cells requires mitogen-activated protein kinase activity

AU - Park, Jung Jae

AU - Moon, Hong Joo

AU - Park, Jin Hyun

AU - Kwon, Taek-Hyun

AU - Park, Youn-Kwan

AU - Kim, Joo-Han

PY - 2016/1/1

Y1 - 2016/1/1

N2 - OBJECTIVE: To determine the role played by mitogen-activated protein kinase (MAPK) signaling in the interactions between macrophages and intervertebral disc (IVD) cells, it was hypothesized that MAPK inhibition would modulate the production of the proinflammatory cytokines associated with inflammatory reaction in IVD cells. METHODS: Human annulus fibrosus (AF) and nucleus pulposus (NP) cells were cocultured with phorbol myristate acetate-stimulated macrophage-like THP-1 cells, with and without SB202190 (a p38-α and -β inhibitor), SP600125 (a c-Jun N-terminal kinase [JNK] inhibitor), and PD98059 (an extracellular signal-regulated kinase [ERK] 1/2 inhibitor). The cytokines in conditioned media from cocultured and macrophage-exposed (nemotic) cells were assayed using enzyme-linked immunosorbent assays (ELISAs). RESULTS: Interleukin (IL)-6 and IL-8 were secreted in greater quantities by the cocultured cells compared with naive IVD cells and macrophages (Mφ) cultured alone. The tumor necrosis factor (TNF)-α and IL-6 levels produced by the NP cells cocultured with Mφs (NP-Mφ) were significantly lower than those produced by AF cells cocultured with Mφs (AF-Mφ). SB202190 dose-dependently suppressed IL-6 secretion by AF-Mφ and NP-Mφ cocultures, and 10 μM SB202190 significantly decreased IL-6 and IL-8 production in nemotic AF and NP pellets. SP600125 at 10 μM significantly suppressed the production of TNFα, IL-6, and IL-8 in AF-Mφ and NP-Mφ cocultures and significantly suppressed IL-1β production in the NP-Mφ coculture. Administration of 10 μM PD98059 significantly decreased IL-6 levels in the AF-Mφ coculture, and decreased the levels of TNFα and IL-8 in both the AF-Mφ and NP-Mφ cocultures. CONCLUSIONS: The present study shows that inhibitors of p38 MAPK effectively controlled IL-6 production during inflammatory reactions and that JNK and ERK1/2 inhibitors successfully suppressed the production of major proinflammatory cytokines during interactions between macrophages and IVD cells. Therefore, selective blockade of these signals may serve as a therapeutic approach to symptomatic IVD degeneration.

AB - OBJECTIVE: To determine the role played by mitogen-activated protein kinase (MAPK) signaling in the interactions between macrophages and intervertebral disc (IVD) cells, it was hypothesized that MAPK inhibition would modulate the production of the proinflammatory cytokines associated with inflammatory reaction in IVD cells. METHODS: Human annulus fibrosus (AF) and nucleus pulposus (NP) cells were cocultured with phorbol myristate acetate-stimulated macrophage-like THP-1 cells, with and without SB202190 (a p38-α and -β inhibitor), SP600125 (a c-Jun N-terminal kinase [JNK] inhibitor), and PD98059 (an extracellular signal-regulated kinase [ERK] 1/2 inhibitor). The cytokines in conditioned media from cocultured and macrophage-exposed (nemotic) cells were assayed using enzyme-linked immunosorbent assays (ELISAs). RESULTS: Interleukin (IL)-6 and IL-8 were secreted in greater quantities by the cocultured cells compared with naive IVD cells and macrophages (Mφ) cultured alone. The tumor necrosis factor (TNF)-α and IL-6 levels produced by the NP cells cocultured with Mφs (NP-Mφ) were significantly lower than those produced by AF cells cocultured with Mφs (AF-Mφ). SB202190 dose-dependently suppressed IL-6 secretion by AF-Mφ and NP-Mφ cocultures, and 10 μM SB202190 significantly decreased IL-6 and IL-8 production in nemotic AF and NP pellets. SP600125 at 10 μM significantly suppressed the production of TNFα, IL-6, and IL-8 in AF-Mφ and NP-Mφ cocultures and significantly suppressed IL-1β production in the NP-Mφ coculture. Administration of 10 μM PD98059 significantly decreased IL-6 levels in the AF-Mφ coculture, and decreased the levels of TNFα and IL-8 in both the AF-Mφ and NP-Mφ cocultures. CONCLUSIONS: The present study shows that inhibitors of p38 MAPK effectively controlled IL-6 production during inflammatory reactions and that JNK and ERK1/2 inhibitors successfully suppressed the production of major proinflammatory cytokines during interactions between macrophages and IVD cells. Therefore, selective blockade of these signals may serve as a therapeutic approach to symptomatic IVD degeneration.

KW - Cytokines

KW - Intervertebral disc degeneration

KW - Mitogen-activated protein kinases

KW - PD98059

KW - SB202190

KW - SP600125

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U2 - 10.3171/2015.3.SPINE14729

DO - 10.3171/2015.3.SPINE14729

M3 - Article

C2 - 26431069

AN - SCOPUS:84974824283

VL - 24

SP - 167

EP - 175

JO - Journal of Neurosurgery: Spine

JF - Journal of Neurosurgery: Spine

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