IL-1β stimulates IL-6 production in cultured skeletal muscle cells through activation of MAP kinase signaling pathway and NF-κB

Guangjo Luo, Dan D. Hershko, Bruce Robb, Curtis J. Wray, Per Olof Hasselgren

Research output: Contribution to journalArticle

79 Citations (Scopus)

Abstract

Recent studies suggest that the skeletal muscle may be a significant site of IL-6 production in various conditions, including exercise, inflammation, hypoperfusion, denervation, and local muscle injury. The mediators and molecular mechanisms regulating muscle IL-6 production are poorly understood. We tested the hypothesis that IL-6 production in muscle cells is regulated by IL-1β and that mitogen-activated protein (MAP) kinase signaling and NF-κB activation are involved in IL-1β-induced IL-6 production. Cultured C2C12 cells, a mouse skeletal muscle cell line, were treated with different concentrations (0.1-2 ng/ml) of IL-1β in the absence or presence of the p38 MAP kinase inhibitor SB-208350 or the p42/44 inhibitor PD-98059. Protein and gene expression of IL-6 were determined by ELISA and real-time PCR, respectively. NF-κB DNA binding activity was determined by electrophoretic mobility shift assay and by transfecting myocytes with a luciferase reporter plasmid containing a promoter construct with multiple repeats of NF-κB binding site. Treatment of myotubes with IL-1β resulted in a dose- and time-dependent increase of IL-6 production accompanied by an ∼25-fold increase in IL-6 mRNA levels. IL-1β stimulated NF-κB DNA binding activity and gene activation. SB-208350 and PD-98059 inhibited the increase in IL-6 production induced by IL-1β. The present results support the concept that skeletal muscle is an important site of IL-6 production. In addition, the results suggest the IL-1β regulates muscle IL-6 production at least in part by activating the MAP kinase pathway and NF-κB.

Original languageEnglish (US)
JournalAmerican Journal of Physiology - Regulatory Integrative and Comparative Physiology
Volume284
Issue number5 53-5
StatePublished - May 1 2003
Externally publishedYes

Fingerprint

Mitogen-Activated Protein Kinases
Interleukin-1
Muscle Cells
Interleukin-6
Skeletal Muscle
Muscle Denervation
Muscles
DNA
Skeletal Muscle Fibers
Electrophoretic Mobility Shift Assay
p38 Mitogen-Activated Protein Kinases
Protein Kinase Inhibitors
Luciferases
Transcriptional Activation
Real-Time Polymerase Chain Reaction
Cultured Cells
Plasmids
Enzyme-Linked Immunosorbent Assay
Binding Sites
Inflammation

Keywords

  • Cytokines
  • Interleukin-6 production
  • Mitogen-activated protein kinase
  • Nuclear factor-κB

ASJC Scopus subject areas

  • Physiology

Cite this

IL-1β stimulates IL-6 production in cultured skeletal muscle cells through activation of MAP kinase signaling pathway and NF-κB. / Luo, Guangjo; Hershko, Dan D.; Robb, Bruce; Wray, Curtis J.; Hasselgren, Per Olof.

In: American Journal of Physiology - Regulatory Integrative and Comparative Physiology, Vol. 284, No. 5 53-5, 01.05.2003.

Research output: Contribution to journalArticle

@article{ede900095bd4429cb336dc0d7ba8ffa4,
title = "IL-1β stimulates IL-6 production in cultured skeletal muscle cells through activation of MAP kinase signaling pathway and NF-κB",
abstract = "Recent studies suggest that the skeletal muscle may be a significant site of IL-6 production in various conditions, including exercise, inflammation, hypoperfusion, denervation, and local muscle injury. The mediators and molecular mechanisms regulating muscle IL-6 production are poorly understood. We tested the hypothesis that IL-6 production in muscle cells is regulated by IL-1β and that mitogen-activated protein (MAP) kinase signaling and NF-κB activation are involved in IL-1β-induced IL-6 production. Cultured C2C12 cells, a mouse skeletal muscle cell line, were treated with different concentrations (0.1-2 ng/ml) of IL-1β in the absence or presence of the p38 MAP kinase inhibitor SB-208350 or the p42/44 inhibitor PD-98059. Protein and gene expression of IL-6 were determined by ELISA and real-time PCR, respectively. NF-κB DNA binding activity was determined by electrophoretic mobility shift assay and by transfecting myocytes with a luciferase reporter plasmid containing a promoter construct with multiple repeats of NF-κB binding site. Treatment of myotubes with IL-1β resulted in a dose- and time-dependent increase of IL-6 production accompanied by an ∼25-fold increase in IL-6 mRNA levels. IL-1β stimulated NF-κB DNA binding activity and gene activation. SB-208350 and PD-98059 inhibited the increase in IL-6 production induced by IL-1β. The present results support the concept that skeletal muscle is an important site of IL-6 production. In addition, the results suggest the IL-1β regulates muscle IL-6 production at least in part by activating the MAP kinase pathway and NF-κB.",
keywords = "Cytokines, Interleukin-6 production, Mitogen-activated protein kinase, Nuclear factor-κB",
author = "Guangjo Luo and Hershko, {Dan D.} and Bruce Robb and Wray, {Curtis J.} and Hasselgren, {Per Olof}",
year = "2003",
month = "5",
day = "1",
language = "English (US)",
volume = "284",
journal = "American Journal of Physiology",
issn = "0193-1857",
publisher = "American Physiological Society",
number = "5 53-5",

}

TY - JOUR

T1 - IL-1β stimulates IL-6 production in cultured skeletal muscle cells through activation of MAP kinase signaling pathway and NF-κB

AU - Luo, Guangjo

AU - Hershko, Dan D.

AU - Robb, Bruce

AU - Wray, Curtis J.

AU - Hasselgren, Per Olof

PY - 2003/5/1

Y1 - 2003/5/1

N2 - Recent studies suggest that the skeletal muscle may be a significant site of IL-6 production in various conditions, including exercise, inflammation, hypoperfusion, denervation, and local muscle injury. The mediators and molecular mechanisms regulating muscle IL-6 production are poorly understood. We tested the hypothesis that IL-6 production in muscle cells is regulated by IL-1β and that mitogen-activated protein (MAP) kinase signaling and NF-κB activation are involved in IL-1β-induced IL-6 production. Cultured C2C12 cells, a mouse skeletal muscle cell line, were treated with different concentrations (0.1-2 ng/ml) of IL-1β in the absence or presence of the p38 MAP kinase inhibitor SB-208350 or the p42/44 inhibitor PD-98059. Protein and gene expression of IL-6 were determined by ELISA and real-time PCR, respectively. NF-κB DNA binding activity was determined by electrophoretic mobility shift assay and by transfecting myocytes with a luciferase reporter plasmid containing a promoter construct with multiple repeats of NF-κB binding site. Treatment of myotubes with IL-1β resulted in a dose- and time-dependent increase of IL-6 production accompanied by an ∼25-fold increase in IL-6 mRNA levels. IL-1β stimulated NF-κB DNA binding activity and gene activation. SB-208350 and PD-98059 inhibited the increase in IL-6 production induced by IL-1β. The present results support the concept that skeletal muscle is an important site of IL-6 production. In addition, the results suggest the IL-1β regulates muscle IL-6 production at least in part by activating the MAP kinase pathway and NF-κB.

AB - Recent studies suggest that the skeletal muscle may be a significant site of IL-6 production in various conditions, including exercise, inflammation, hypoperfusion, denervation, and local muscle injury. The mediators and molecular mechanisms regulating muscle IL-6 production are poorly understood. We tested the hypothesis that IL-6 production in muscle cells is regulated by IL-1β and that mitogen-activated protein (MAP) kinase signaling and NF-κB activation are involved in IL-1β-induced IL-6 production. Cultured C2C12 cells, a mouse skeletal muscle cell line, were treated with different concentrations (0.1-2 ng/ml) of IL-1β in the absence or presence of the p38 MAP kinase inhibitor SB-208350 or the p42/44 inhibitor PD-98059. Protein and gene expression of IL-6 were determined by ELISA and real-time PCR, respectively. NF-κB DNA binding activity was determined by electrophoretic mobility shift assay and by transfecting myocytes with a luciferase reporter plasmid containing a promoter construct with multiple repeats of NF-κB binding site. Treatment of myotubes with IL-1β resulted in a dose- and time-dependent increase of IL-6 production accompanied by an ∼25-fold increase in IL-6 mRNA levels. IL-1β stimulated NF-κB DNA binding activity and gene activation. SB-208350 and PD-98059 inhibited the increase in IL-6 production induced by IL-1β. The present results support the concept that skeletal muscle is an important site of IL-6 production. In addition, the results suggest the IL-1β regulates muscle IL-6 production at least in part by activating the MAP kinase pathway and NF-κB.

KW - Cytokines

KW - Interleukin-6 production

KW - Mitogen-activated protein kinase

KW - Nuclear factor-κB

UR - http://www.scopus.com/inward/record.url?scp=18244421931&partnerID=8YFLogxK

UR - http://www.scopus.com/inward/citedby.url?scp=18244421931&partnerID=8YFLogxK

M3 - Article

C2 - 12676746

AN - SCOPUS:18244421931

VL - 284

JO - American Journal of Physiology

JF - American Journal of Physiology

SN - 0193-1857

IS - 5 53-5

ER -