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Papers In Press, published online ahead of print April 8, 2002
Institute of Biotechnology in Medicine, National Yang-Ming University, Taipei 112
Corresponding Author: hyhsu{at}ym.edu.tw
Lipopolysaccharide (LPS) stimulates macrophages to release inflammatory cytokines, interleukin-1b (IL-1) and TNF. LPS-induced TNF suppresses scavenger receptor functions in macrophages (van Lenten, B.J. and Fogelman, A.M. (1992) J. Immunol. 148, 112-116), which is regulated by TNF-mediated protein kinases (Hsu, H.Y. and Twu, Y.C. (2000) J. Biol. Chem. 275, 41035-41048). To examine the molecular mechanism for LPS induction of IL-1 in macrophages, we demonstrate that LPS quickly stimulated reactive oxygen species (ROS), and 3 h later induced prointerleukin-1b (proIL-1, precursor of IL-1) production and IL-1 secretion. LPS stimulated proIL-1 message/protein between 3 h and 10 h; however, there was a 40% reduction of proIL-1 in pre-incubation of the antioxidant, N-acetyl-cysteine (NAC). Moreover, NAC moderated LPS-induced IL-1 secretion partially via ICE. The maximal activity of LPS-induced ERK, JNK and p38 was 12-fold (30 min), 5-fold (30 min) and 16-fold (15 min), respectively. In contrast, NAC reduced ERK activity to 60% and decreased p38 activity to the basal level, but JNK activity was induced 2-fold. Furthermore, the pharmacological antagonists LY294002, SB203580, curcumin, calphostin C and PD98059 revealed the diverse roles of LPS-mediated protein kinases in proIL-1. On the other hand, NAC and diphenyleneiodonium chloride partially inhibited LPS-induced Rac activity and protein tyrosine kinase (PTK), indicating that LPS-mediated ROS and NADPH oxidase correspond to Rac activation and IL-1 expression. Our findings establish for the first time that LPS-mediated PTK/PI-3 kinase/Rac/p38 pathways play a more important role than pathways of PTK/PKC/MEK/ERK and of PTK/PI-3 kinase/Rac/JNK in the regulation of proIL-1/IL-1. The findings also further elucidate the critical role of LPS-mediated ROS in signal transduction pathways. Our results suggest that understanding LPS-transduced signals in IL-1 induction upon the antibacterial action of macrophages should provide a therapeutic strategy for aberrant inflammatory responses leading to severe cellular injury or concurrent multi-organ septic damage.
J. Biol. Chem, 10.1074/jbc.M111883200
Submitted on December 13, 2001
Revised on March 22, 2002
Accepted on April 5, 2002
Lipopolysaccharide-mediated reactive oxygen species and signal transduction in the regulation of IL-1 gene expression
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