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J. Biol. Chem., Vol. 277, Issue 34, 30551-30558, August 23, 2002
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From the Department of Gastroenterology, Research Center,
Universidad Complutense Hospital Universitario "12 de Octubre,"
28041-Madrid, Spain
Malondialdehyde, the end product of lipid
peroxidation, has been shown to stimulate collagen
Sp1 and Sp3 Transcription Factors Mediate Malondialdehyde-induced
Collagen
1(I) Gene Expression in Cultured Hepatic Stellate
Cells*
1(I)
(Col1a1) gene expression. However, mechanisms of this effect are
unclear. The purpose of this study was to clarify these mechanisms. Rat
hepatic stellate cells were cultured in the presence of 200 µM malondialdehyde, and the effects on collagen
gene expression and the binding of nuclear proteins to the
col1a1 promoter were analyzed. Malondialdehyde treatment induced an increase in the cellular levels of
col1a1 mRNA that was abrogated by pretreating cells
with cycloheximide, p-hydroxymercuribenzoate,
pyridoxal 5'-phosphate, and mithramycin. Transient transfections showed
that malondialdehyde exerted its effect through regulatory elements
located between 
220 and 110 bp of the col1a1 promoter.
Gel retardation assays demonstrated that malondialdehyde increased the
binding of nuclear proteins to two elements located between 161 and
110 bp of the col1a1 promoter. These bindings were
supershifted with Sp1 and Sp3 antibodies. Finally, malondialdehyde
increased cellular levels of the Sp1 and Sp3 proteins and
Sp1 mRNA. Our data indicated that treatment of hepatic
stellate cells with malondialdehyde stimulated col1a1 gene
expression by inducing the synthesis of Sp1 and Sp3 and their binding
to two regulatory elements located between 161 and 110 bp of the
col1a1 promoter.
*
This work was supported in part by Grants 00/373 and 01/1447
from the "Fondo de Investigación Sanitaria," Spain, and by
Grant 08.2/0047/2001.1 from the "Comunidad de Madrid," Spain.The costs of publication of this
article were defrayed in part by the
payment of page charges. The article
must therefore be hereby marked
"advertisement" in
accordance with 18 U.S.C. Section
1734 solely to indicate this fact.
To whom correspondence should be addressed: Servicio de Aparato
Digestivo, Hospital Universitario "12 de Octubre,"
c/Andalucía, Km 5,400, 28041-Madrid, Spain. Tel.:
34-91-390-8020; Fax: 34-91-390-8280; E-mail: jasolis@h12o.es and
jsolis@hdoc.insalud.es.
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