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IL-1beta induces MMP-9 expression via a Ca2+-dependent CaMKII/JNK/c-JUN cascade in rat brain astrocytes

Cheng-Ying Wu1, Hsi-Lung Hsieh, Chi-Chin Sun

  • 1Department of Physiology and Pharmacology, Chang Gung University, Kwei-San, Tao-Yuan, Taiwan.

Glia
|May 21, 2009
PubMed

Insights

Interleukin-1beta induces matrix metalloproteinase-9 expression in rat brain astrocytes via a calcium-dependent pathway involving CaMKII, JNK, and c-Jun activation. This signaling cascade promotes MMP-9 transcription and cellular expression.

Area of Science:

  • Neuroscience
  • Cell Biology
  • Molecular Biology

Background:

  • Interleukin (IL)-1beta is known to induce matrix metalloproteinase (MMP)-9 expression through mitogen-activated protein kinases (MAPKs) like JNK in rat brain astrocyte-1 (RBA-1) cells.
  • The precise role of Ca(2+)-dependent Ca(2+)/calmodulin-dependent protein kinase II (CaMKII) in mediating JNK activation and subsequent MMP-9 expression by IL-1beta remains unclear.

Purpose of the Study:

  • To elucidate the signaling pathway through which IL-1beta induces MMP-9 expression in RBA-1 cells, focusing on the involvement of Ca(2+)/CaMKII/JNK/c-Jun.
  • To investigate the mechanism of transcription factor recruitment to the MMP-9 promoter.

Main Methods:

  • Zymography, Western blotting, and RT-PCR were used to assess MMP-9 expression.
  • Intracellular calcium ([Ca(2+)](i)) levels and protein phosphorylation (CaMKII, JNK1/2) were measured.
  • Inhibitors (BAPTA, thapsigargin, KN-62, SP600125), short hairpin RNA (shRNA) for c-Jun and CaMKII, and promoter-luciferase reporter assays were employed.
  • Immunoprecipitation and chromatin immunoprecipitation (ChIP)-PCR assays were performed to analyze transcription factor binding.

Main Results:

  • IL-1beta-induced MMP-9 expression was significantly attenuated by Ca(2+) chelators, ER Ca(2+)-ATPase inhibitors, and specific inhibitors of CaMKII and JNK.
  • IL-1beta increased intracellular Ca(2+) and phosphorylation of CaMKII and JNK1/2, effects blocked by upstream inhibitors.
  • Knockdown of c-Jun or CaMKII, and inhibition of JNK, blocked MMP-9 upregulation.
  • IL-1beta stimulated the recruitment of c-Jun to AP-1 binding sites in the MMP-9 promoter, a process dependent on Ca(2+)/CaMKII/JNK signaling.

Conclusions:

  • The Ca(2+)/CaMKII/JNK/c-Jun pathway is critical for IL-1beta-induced MMP-9 expression in RBA-1 cells.
  • IL-1beta activates MMP-9 transcription through the Ca(2+)-dependent activation of JNK/c-Jun and subsequent AP-1 recruitment to the MMP-9 promoter.
  • These findings reveal a novel signaling cascade regulating MMP-9 expression in astrocytes.

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