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J Biol Chem, Vol. 274, Issue 4, 2234-2242, January 22, 1999
From the Laboratory of Food and Biodynamics, In the present study, we studied the
signal transduction mechanism that is involved in the expression of
c-Jun protein evident after exposure of rat liver epithelial RL34 cells
to the major end product of oxidized fatty acid metabolism,
4-hydroxy-2-nonenal (HNE). HNE treatment of the cells resulted in
depletion of intracellular glutathione (GSH) and in the formation of
protein-bound HNE in plasma membrane. In addition, HNE strongly induced
intracellular peroxide production, suggesting that HNE exerted
oxidative stress on the cells. Potent expression of c-Jun occurred
within 30 min of HNE treatment, which was accompanied by a
time-dependent increase in activator protein-1 (AP-1) DNA
binding activity. We found that HNE caused an immediate increase in
tyrosine phosphorylation in RL34 cells. In addition, HNE strongly
induced phosphorylation of c-Jun N-terminal kinases (JNK) and p38
mitogen-activated protein kinases and also moderately induced
phosphorylation of extracellular signal-regulated kinases. The
phosphorylation of JNK was accompanied by a rapid and transient
increase in JNK and p38 activities, whereas changes in the activity of
extracellular signal-regulated kinase were scarcely observed. GSH
depletion by
L-buthionine-S,R-sulfoximine, a specific inhibitor of GSH biosynthesis, only slightly enhanced peroxide production and JNK activation, suggesting that HNE exerted these effects independent of GSH depletion. This and the findings that
(i) HNE strongly induced intracellular peroxide production, (ii)
HNE-induced JNK activation was inhibited by pretreatment of the cells
with a thiol antioxidant, N-acetylcysteine, and (iii) H2O2 significantly activated JNK support the
hypothesis that pro-oxidants play a crucial role in the HNE-induced
activation of stress signaling pathways. In addition, we found that,
among the inhibitors of tyrosine kinases, cyclooxygenase, and
Ca2+ influx, only quercetin exerted a significant
inhibitory effect on HNE-induced JNK activation. In light of the
JNK-dependent induction of c-jun transcription
and the AP-1-induced transcription of xenobiotic-metabolizing enzymes,
these data may show a potential critical role for JNK in the induction
of a cellular defense program against toxic products generated from
lipid peroxidation.
Copyright © 1999 by The American Society for Biochemistry and Molecular Biology, Inc.
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J.-Z. Cheng, R. Sharma, Y. Yang, S. S. Singhal, A. Sharma, M. K. Saini, S. V. Singh, P. Zimniak, S. Awasthi, and Y. C. Awasthi Accelerated Metabolism and Exclusion of 4-Hydroxynonenal through Induction of RLIP76 and hGST5.8 Is an Early Adaptive Response of Cells to Heat and Oxidative Stress J. Biol. Chem., October 26, 2001; 276(44): 41213 - 41223. [Abstract] [Full Text] [PDF]
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