Andrographolide enhances nuclear factor-κB subunit p65 Ser 536 dephosphorylation through activation of protein phosphatase 2A in vascular smooth muscle cells

Cheng Y. Hsieh, Ming J. Hsu, George Hsiao, Yi H. Wang, Chi W. Huang, Shiuan W. Chen, Thanasekaran Jayakumar, Pei T. Chiu, Yi H. Chiu, Joen R. Sheu

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Abstract

Recent studies have demonstrated that transcription factor nuclear factor (NF)-κB inhibition may contribute to the protective anti-inflammatory actions of andrographolide, an abundant component of plants of the genus Andrographis. However, the precise mechanism by which andrographolide inhibits NF-κB signaling remains unclear. We thus investigated the mechanism involved in andrographolide suppression of NF-κB signaling in rat vascular smooth muscle cells (VSMCs) exposed to proinflammatory stimuli, LPS, and IFN-γ. Andrographolide was shown to suppress LPS/IFN-γ-induced inducible nitric-oxide synthase and matrix metalloprotease 9 expression in rat VSMCs. Andrographolide also inhibited LPS/IFN-γ-induced p65 nuclear translocation, DNA binding activity, p65 Ser536 phosphorylation, and NF-κB reporter activity. However, IKK phosphorylation and downstream inhibitory κBα phosphorylation and degradation were not altered by the presence of andrographolide in LPS/IFN-γ-stimulated VSMCs. These andrographolide inhibitory actions could be prevented by selective inhibition of neutral sphingomyelinase and protein phosphatase 2A (PP2A). Furthermore, andrographolide was demonstrated to increase ceramide formation and PP2A activity in VSMCs and to inhibit neointimal formation in rat carotid injury models. These results suggest that andrographolide caused neutral sphingomyelinase-mediated ceramide formation and PP2A activation to dephosphorylate p65 Ser536, leading to NF-κB inactivation and subsequent inducible nitric-oxide synthase down-regulation in rat VSMCs stimulated by LPS and IFN-γ.

Original languageEnglish
Pages (from-to)5942-5955
Number of pages14
JournalJournal of Biological Chemistry
Volume286
Issue number8
DOIs
Publication statusPublished - Feb 25 2011

Fingerprint

Protein Phosphatase 2
Vascular Smooth Muscle
Smooth Muscle Myocytes
Muscle
Chemical activation
Cells
Phosphorylation
Rats
Sphingomyelin Phosphodiesterase
Ceramides
Nitric Oxide Synthase Type II
Andrographis
Plant Structures
andrographolide
Metalloproteases
Anti-Inflammatory Agents
Transcription Factors
Down-Regulation
Degradation

ASJC Scopus subject areas

  • Biochemistry
  • Cell Biology
  • Molecular Biology

Cite this

Andrographolide enhances nuclear factor-κB subunit p65 Ser 536 dephosphorylation through activation of protein phosphatase 2A in vascular smooth muscle cells. / Hsieh, Cheng Y.; Hsu, Ming J.; Hsiao, George; Wang, Yi H.; Huang, Chi W.; Chen, Shiuan W.; Jayakumar, Thanasekaran; Chiu, Pei T.; Chiu, Yi H.; Sheu, Joen R.

In: Journal of Biological Chemistry, Vol. 286, No. 8, 25.02.2011, p. 5942-5955.

Research output: Contribution to journalArticle

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abstract = "Recent studies have demonstrated that transcription factor nuclear factor (NF)-κB inhibition may contribute to the protective anti-inflammatory actions of andrographolide, an abundant component of plants of the genus Andrographis. However, the precise mechanism by which andrographolide inhibits NF-κB signaling remains unclear. We thus investigated the mechanism involved in andrographolide suppression of NF-κB signaling in rat vascular smooth muscle cells (VSMCs) exposed to proinflammatory stimuli, LPS, and IFN-γ. Andrographolide was shown to suppress LPS/IFN-γ-induced inducible nitric-oxide synthase and matrix metalloprotease 9 expression in rat VSMCs. Andrographolide also inhibited LPS/IFN-γ-induced p65 nuclear translocation, DNA binding activity, p65 Ser536 phosphorylation, and NF-κB reporter activity. However, IKK phosphorylation and downstream inhibitory κBα phosphorylation and degradation were not altered by the presence of andrographolide in LPS/IFN-γ-stimulated VSMCs. These andrographolide inhibitory actions could be prevented by selective inhibition of neutral sphingomyelinase and protein phosphatase 2A (PP2A). Furthermore, andrographolide was demonstrated to increase ceramide formation and PP2A activity in VSMCs and to inhibit neointimal formation in rat carotid injury models. These results suggest that andrographolide caused neutral sphingomyelinase-mediated ceramide formation and PP2A activation to dephosphorylate p65 Ser536, leading to NF-κB inactivation and subsequent inducible nitric-oxide synthase down-regulation in rat VSMCs stimulated by LPS and IFN-γ.",
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AU - Hsu, Ming J.

AU - Hsiao, George

AU - Wang, Yi H.

AU - Huang, Chi W.

AU - Chen, Shiuan W.

AU - Jayakumar, Thanasekaran

AU - Chiu, Pei T.

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AU - Sheu, Joen R.

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