Fb. Riquet et al., Suppression of type I collagen gene expression by prostaglandins in fibroblasts is mediated at the transcriptional level, MOL MED, 6(8), 2000, pp. 705-719
Citations number
68
Categorie Soggetti
Research/Laboratory Medicine & Medical Tecnology","Medical Research General Topics
Background: Tissues undergoing a chronic inflammatory process, such as the
synovium in rheumatoid arthritis, are characterized by the infiltration of
lymphocytes of different subsets and activation of monocyte/macrophages. In
terleultin-1 (IL-1), a monocyte/macrophage product that stimulates synovial
fibroblasts to produce matrix metalloproteinases (MMPs), prostaglandins, a
nd other cytokines, also has profound effects on the synthesis of extracell
ular matrix components such as type I collagen. In previous studies, we hav
e shown that synovial fibroblasts and chondrocytes isolated from human join
t tissues are particularly sensitive to prostaglandins, which modulate the
effects of IL-1 on collagen gene expression in an autocrine manner.
Materials and Methods: BALBc/3T3 fibroblasts were treated with IL-I and pro
staglandins in the absence and presence of indomethacin to inhibit endogeno
us prostaglandin biosynthesis. Collagen synthesis was analyzed by SDS-PAGE
as [H-3]proline-labeled, secreted proteins, and prostaglandin production an
d cyclic adenosine 3',5'-cyclic monophosphate (camp) content were assayed.
The expression of type I collagen gene (Col1a1) promoter-reporter gene cons
tructs was examined in transient transfection experiments, and the binding
of nuclear factors to the Col1a1 promoter region spanning -222 bp/+ 116 bp
was analyzed by DNase I footprinting and electrophoretic mobility shift (EM
SA) assays.
Results: IL-1 increased the synthesis of type I and type III collagens in B
ALBc/3T3 fibroblasts; greater increases were observed when IL-l-stimulated
synthesis of PGE, was blocked by indomethacin. Transient transfection exper
iments demonstrated dose-dependent inhibition of the -222 bp Col1a1 promote
r by exogenously added prostaglandins with the order of potency of PGF(2)al
pha > PGE(2) > PGE(1). DNase I footprinting showed increased protection, wh
ich extended from the region immediately upstream of the TATA box, owing to
the binding of nuclear factors from PGE(2)- or PGE(1)-treated BALBc/3T3 ce
lls. EMSA analysis showed zinc-dependent differences in the binding of nucl
ear factors from unheated and prostaglandin-treated cells to the -84 bp/-29
bp region of the Col1a1 promoter. conclusions: These results show that the
inhibition of Col1a1 expression by IL-1 in fibroblasts is mediated by pros
taglandins at the transcriptional level and suggest that PGE-responsive fac
tors may interact directly or indirectly with basal regulatory elements in
the proximal promoter region of the Col1a1 gene.