C-Src-dependent MAPKs/AP-1 activation is involved in TNF-α-induced matrix metalloproteinase-9 expression in rat heart-derived H9c2 cells

Chuen Mao Yang, I-Ta Lee, Chih Chung Lin, Chao Hung Wang, Wen Jin Cherng, Li Der Hsiao

Research output: Contribution to journalArticle

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Abstract

TNF-α plays a critical mediator in the pathogenesis of chronic heart failure contributing to cardiac remodeling and peripheral vascular disturbances. The implication of TNF-α in inflammatory responses has been shown to be mediated through up-regulation of inflammatory genes, including matrix metalloproteinase-9 (MMP-9). However, the detailed mechanisms of TNF-α-induced MMP-9 expression are largely unclear in the heart cells. Here, we demonstrated that in rat embryonic-heart derived H9c2 cells, TNF-α could induce MMP-9 mRNA expression associated with an increase in the secretion of MMP-9, determined by real-time PCR, zymography, and promoter activity assays. TNF-α-mediated responses were attenuated by pretreatment with the inhibitor of c-Src (PP1), EGFR (AG1478), PDGFR (AG1296), PI3K (LY294002), Akt (SH-5), MEK1/2 (U0126), p38 MAPK (SB202190), JNK1/2 (SP600125), or AP-1 (Tanshinone IIA) and transfection with siRNA of c-Src, EGFR, PDGFR, p110, Akt, or c-Jun. TNF-α stimulated c-Src, PDGFR, and EGFR phosphorylation, which were reduced by PP1. In addition, TNF-α-stimulated Akt phosphorylation was inhibited by PP1, AG1478, AG1296, or LY294002. We further demonstrated that TNF-α markedly stimulated p38 MAPK, p42/p44 MAPK, and JNK1/2 phosphorylation via a c-Src/EGFR, PDGFR/PI3 K/Akt pathway. Finally, we showed that, in H9c2 cells, TNF-α-stimulated AP-1 promoter activity, c-Jun mRNA expression, and c-Jun phosphorylation were attenuated by PP1, AG1478, AG1296, LY294002, SB202190, SP600125, or U0126. These results suggested that TNF-α-induced MMP-9 expression is mediated through a c-Src/EGFR, PDGFR/PI3K/Akt/MAPKs/AP-1 cascade in H9c2 cells. Consequently, MMP-9 induction may contribute to cell migration and cardiovascular inflammation.

Original languageEnglish
Pages (from-to)1115-1123
Number of pages9
JournalBiochemical Pharmacology
Volume85
Issue number8
DOIs
Publication statusPublished - Apr 15 2013
Externally publishedYes

Fingerprint

Matrix Metalloproteinase 9
Transcription Factor AP-1
Rats
Phosphorylation
Chemical activation
2-(4-morpholinyl)-8-phenyl-4H-1-benzopyran-4-one
p38 Mitogen-Activated Protein Kinases
Phosphatidylinositol 3-Kinases
Messenger RNA
Mitogen-Activated Protein Kinase 3
Mitogen-Activated Protein Kinase 1
Small Interfering RNA
Cell Movement
Transfection
Real-Time Polymerase Chain Reaction
Assays
Up-Regulation
Heart Failure
Genes
Inflammation

Keywords

  • Cytokine
  • Matrix metalloproteinase-9
  • Transcription factor
  • Tyrosine kinase

ASJC Scopus subject areas

  • Biochemistry
  • Pharmacology

Cite this

C-Src-dependent MAPKs/AP-1 activation is involved in TNF-α-induced matrix metalloproteinase-9 expression in rat heart-derived H9c2 cells. / Yang, Chuen Mao; Lee, I-Ta; Lin, Chih Chung; Wang, Chao Hung; Cherng, Wen Jin; Hsiao, Li Der.

In: Biochemical Pharmacology, Vol. 85, No. 8, 15.04.2013, p. 1115-1123.

Research output: Contribution to journalArticle

Yang, Chuen Mao ; Lee, I-Ta ; Lin, Chih Chung ; Wang, Chao Hung ; Cherng, Wen Jin ; Hsiao, Li Der. / C-Src-dependent MAPKs/AP-1 activation is involved in TNF-α-induced matrix metalloproteinase-9 expression in rat heart-derived H9c2 cells. In: Biochemical Pharmacology. 2013 ; Vol. 85, No. 8. pp. 1115-1123.
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AU - Cherng, Wen Jin

AU - Hsiao, Li Der

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