Tumor necrosis factor ? down-regulates expression of the ?1(I) collagen gene in rat hepatic stellate cells through a p20C/EBP?- and C/EBP?-dependent mechanism

Tumor necrosis factor ? (TNF-?) is one of the key cytokines of the acute phase response and of many inflammatory processes. This cytokine has several antifibrogenic actions and down-regulates the expression of the type I collagen genes and induces the expression of metalloproteinases. Because TNF-? directly antagonizes some fibrogenic actions of transforming growth factor ?1 (TGF-?1), we considered it important to map the cis-acting regulatory element of the ?1(I) collagen (col1a1) promoter involved in TNF- ? responsiveness in hepatic stellate cells (HSC), to investigate the transcription factors that bind to it, and to establish possible mechanisms by which TNF-? downregulates its expression. In this article, we show the presence of a functional TNF-?-responsive element (TaRE) in the -378 to -345 region of the col1a1 promoter. This element colocalizes with a previously reported TGF-?1-responsive element. We further demonstrate that TNF-? induces nuclear translocation and binding of transcriptional complexes containing p20C/EBP?, p35C/EBP?, and C/EBP? to this sequence of the promoter. Transient overexpression of C/EBP? or p20C/EBP?, the natural dominant negative form of C/EBP? in HSC, down-regulated activity of a CAT reporter vector driven by -412 to +110 of the col1a1 promoter. Taken together, these data suggest that the -378 to -340 region of the col1a1 promoter is the site of convergence of different stimuli that ultimately modulate col1a1 gene transcription