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-stimulated
sustained ERK activation and MMP-9 induction
Department of Anatomy and Cell Biology, The University of Iowa College of Medicine, Iowa City, Iowa 52242
We have recently demonstrated
that interleukin-1
(IL-1) stimulates matrix metalloproteinase-9
(MMP-9) induction. In this study we have investigated the roles of
superoxide and extracellular signal-regulated kinase (ERK) activation
in MMP-9 induction following exposure to IL-1. IL-1 stimulated
biphasic ERK activation in vascular smooth muscle (VSM) cells, a
transient activation that reached a maximum at 15 min and declined to
baseline levels within 1 h, and a second phase of sustained ERK
activation lasting up to 8 h. To determine the role of ERK in
IL-1-stimulated MMP-9 induction, we treated cells with the specific
ERK pathway inhibitor PD-98059 at different time intervals after
IL-1 stimulation. Addition of PD-98059 up to 4 h after IL-1
stimulation significantly inhibited MMP-9 induction, suggesting a role
for sustained ERK activation in MMP-9 induction. IL-1 treatment
stimulated superoxide production in VSM cells that was inhibited by
pretreatment of cells with the superoxide scavenger
N-acetyl-L-cysteine (NAC) and also by
overexpression of the human manganese superoxide dismutase (MnSOD)
gene. Treatment of VSM cells with NAC selectively inhibited the
sustained phase of ERK activation without influencing the transient
phase, suggesting a role for reactive oxygen species in sustained ERK
activation. In addition, both NAC treatment and MnSOD overexpression
significantly inhibited IL-1-stimulated MMP-9 induction
(P < 0.05). The results demonstrate that
IL-1-dependent MMP-9 induction is mediated by superoxide-stimulated
ERK activation.
matrix metalloproteinases; extracellular signal-regulated kinase; vascular smooth muscle cells; gene transfer; N-acetyl-L-cysteine; superoxide dismutase
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