Abstract

This study investigates the signaling pathway involved in inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) release caused by Staphylococcus aureus lipoteichoic acid (LTA) in RAW 264.7 macrophages. A phosphatidylcholine-phospholipase C (PC-PLC) inhibitor (D-609) and a phosphatidylinositol-phospholipase C (PI-PLC) inhibitor (U-73122) attenuated LTA-induced iNOS expression and NO release. Two PKC inhibitors (Go 6976 and Ro 31-8220), an NF-κB inhibitor (pyrrolidine dithiocarbamate; PDTC), and long-term (24 h) 12-phorbol-13-myristate acetate (PMA) treatment each also inhibited LTA-induced iNOS expression and NO release. Treatment of cells with LTA caused an increase in PKC activity; this stimulatory effect was inhibited by D-609, U-73122, or Ro 31-8220. Stimulation of cells with LTA caused IκB-α phosphorylation and IκB-α degradation in the cytosol, and translocation of p65 and p50 NF-κB from the cytosol to the nucleus. Treatment of cells with LTA caused NF-κB activation by detecting the formation of NF-κB-specific DNA-protein complexes in the nucleus; this effect was inhibited by Go 6976, Ro 31-8220, long-term PMA treatment, PDTC, L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK), and calpain inhibitor I. These results suggest that LTA might activate PC-PLC and PI-PLC to induce PKC activation, which in turn initiates NF-κB activation, and finally induces iNOS expression and NO release in RAW 264.7 macrophages.

Original languageEnglish
Pages (from-to)136-145
Number of pages10
JournalJournal of Biomedical Science
Volume10
Issue number1
DOIs
Publication statusPublished - 2003

Fingerprint

Macrophages
Nitric Oxide Synthase
Protein Kinase C
Staphylococcus aureus
Type C Phospholipases
Nitric Oxide Synthase Type II
Nitric Oxide
Chemical activation
Myristic Acid
Tetradecanoylphorbol Acetate
Phosphatidylinositols
Phosphatidylcholines
Cytosol
Acetates
Cells
Phosphorylation
lipoteichoic acid
Ketones
Degradation
DNA

Keywords

  • Inducible nitric oxide synthase
  • Lipoteichoic acid
  • NF-κB
  • Nitric oxide
  • Protein kinase C
  • RAW 264.7 macrophages

ASJC Scopus subject areas

  • Biochemistry, Genetics and Molecular Biology(all)

Cite this

@article{5df0359314324a5c9b77995ad22ba9ec,
title = "Induction of nitric oxide synthase in RAW 264.7 macrophages by lipoteichoic acid from Staphylococcus aureus: Involvement of protein kinase C- and nuclear factor-κB-dependent mechanisms",
abstract = "This study investigates the signaling pathway involved in inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) release caused by Staphylococcus aureus lipoteichoic acid (LTA) in RAW 264.7 macrophages. A phosphatidylcholine-phospholipase C (PC-PLC) inhibitor (D-609) and a phosphatidylinositol-phospholipase C (PI-PLC) inhibitor (U-73122) attenuated LTA-induced iNOS expression and NO release. Two PKC inhibitors (Go 6976 and Ro 31-8220), an NF-κB inhibitor (pyrrolidine dithiocarbamate; PDTC), and long-term (24 h) 12-phorbol-13-myristate acetate (PMA) treatment each also inhibited LTA-induced iNOS expression and NO release. Treatment of cells with LTA caused an increase in PKC activity; this stimulatory effect was inhibited by D-609, U-73122, or Ro 31-8220. Stimulation of cells with LTA caused IκB-α phosphorylation and IκB-α degradation in the cytosol, and translocation of p65 and p50 NF-κB from the cytosol to the nucleus. Treatment of cells with LTA caused NF-κB activation by detecting the formation of NF-κB-specific DNA-protein complexes in the nucleus; this effect was inhibited by Go 6976, Ro 31-8220, long-term PMA treatment, PDTC, L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK), and calpain inhibitor I. These results suggest that LTA might activate PC-PLC and PI-PLC to induce PKC activation, which in turn initiates NF-κB activation, and finally induces iNOS expression and NO release in RAW 264.7 macrophages.",
keywords = "Inducible nitric oxide synthase, Lipoteichoic acid, NF-κB, Nitric oxide, Protein kinase C, RAW 264.7 macrophages",
author = "Kuo, {Chen Tzu} and Chiang, {Ling Ling} and Chun-Nin Lee and Yu, {Ming Chih} and Bai, {Kuan Jen} and Lee, {Horng Mo} and Lee, {Wen Sen} and Sheu, {Joen Rong} and Lin, {Chien Huang}",
year = "2003",
doi = "10.1159/000068076",
language = "English",
volume = "10",
pages = "136--145",
journal = "Journal of Biomedical Science",
issn = "1021-7770",
publisher = "BioMed Central",
number = "1",

}

TY - JOUR

T1 - Induction of nitric oxide synthase in RAW 264.7 macrophages by lipoteichoic acid from Staphylococcus aureus

T2 - Involvement of protein kinase C- and nuclear factor-κB-dependent mechanisms

AU - Kuo, Chen Tzu

AU - Chiang, Ling Ling

AU - Lee, Chun-Nin

AU - Yu, Ming Chih

AU - Bai, Kuan Jen

AU - Lee, Horng Mo

AU - Lee, Wen Sen

AU - Sheu, Joen Rong

AU - Lin, Chien Huang

PY - 2003

Y1 - 2003

N2 - This study investigates the signaling pathway involved in inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) release caused by Staphylococcus aureus lipoteichoic acid (LTA) in RAW 264.7 macrophages. A phosphatidylcholine-phospholipase C (PC-PLC) inhibitor (D-609) and a phosphatidylinositol-phospholipase C (PI-PLC) inhibitor (U-73122) attenuated LTA-induced iNOS expression and NO release. Two PKC inhibitors (Go 6976 and Ro 31-8220), an NF-κB inhibitor (pyrrolidine dithiocarbamate; PDTC), and long-term (24 h) 12-phorbol-13-myristate acetate (PMA) treatment each also inhibited LTA-induced iNOS expression and NO release. Treatment of cells with LTA caused an increase in PKC activity; this stimulatory effect was inhibited by D-609, U-73122, or Ro 31-8220. Stimulation of cells with LTA caused IκB-α phosphorylation and IκB-α degradation in the cytosol, and translocation of p65 and p50 NF-κB from the cytosol to the nucleus. Treatment of cells with LTA caused NF-κB activation by detecting the formation of NF-κB-specific DNA-protein complexes in the nucleus; this effect was inhibited by Go 6976, Ro 31-8220, long-term PMA treatment, PDTC, L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK), and calpain inhibitor I. These results suggest that LTA might activate PC-PLC and PI-PLC to induce PKC activation, which in turn initiates NF-κB activation, and finally induces iNOS expression and NO release in RAW 264.7 macrophages.

AB - This study investigates the signaling pathway involved in inducible nitric oxide synthase (iNOS) expression and nitric oxide (NO) release caused by Staphylococcus aureus lipoteichoic acid (LTA) in RAW 264.7 macrophages. A phosphatidylcholine-phospholipase C (PC-PLC) inhibitor (D-609) and a phosphatidylinositol-phospholipase C (PI-PLC) inhibitor (U-73122) attenuated LTA-induced iNOS expression and NO release. Two PKC inhibitors (Go 6976 and Ro 31-8220), an NF-κB inhibitor (pyrrolidine dithiocarbamate; PDTC), and long-term (24 h) 12-phorbol-13-myristate acetate (PMA) treatment each also inhibited LTA-induced iNOS expression and NO release. Treatment of cells with LTA caused an increase in PKC activity; this stimulatory effect was inhibited by D-609, U-73122, or Ro 31-8220. Stimulation of cells with LTA caused IκB-α phosphorylation and IκB-α degradation in the cytosol, and translocation of p65 and p50 NF-κB from the cytosol to the nucleus. Treatment of cells with LTA caused NF-κB activation by detecting the formation of NF-κB-specific DNA-protein complexes in the nucleus; this effect was inhibited by Go 6976, Ro 31-8220, long-term PMA treatment, PDTC, L-1-tosylamido-2-phenylethyl chloromethyl ketone (TPCK), and calpain inhibitor I. These results suggest that LTA might activate PC-PLC and PI-PLC to induce PKC activation, which in turn initiates NF-κB activation, and finally induces iNOS expression and NO release in RAW 264.7 macrophages.

KW - Inducible nitric oxide synthase

KW - Lipoteichoic acid

KW - NF-κB

KW - Nitric oxide

KW - Protein kinase C

KW - RAW 264.7 macrophages

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U2 - 10.1159/000068076

DO - 10.1159/000068076

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VL - 10

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EP - 145

JO - Journal of Biomedical Science

JF - Journal of Biomedical Science

SN - 1021-7770

IS - 1

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