Regulatory Effect of Extracellular Signal-Regulated Kinases (ERK) on Type I Collagen Synthesis in Human Dermal Fibroblasts Stimulated by IL-4 and IL-13

Abstract

Synthesis of collagen is up-regulated by pro-fibrogenic growth factors and cytokines such as TGF-β 1, IL-4, and IL-13 binding to their corresponding cell membrane receptors of fibroblasts. The ERK pathway is an important MAPK signaling pathway that is involved in regulating cell function. The aim of our studies was to examine effects of IL-4 and IL-13 on the ERK signaling pathway and its function in regulating type I collagen gene expression in human fibroblasts. We found that human dermal fibroblasts treated with IL-4 and IL-13 exhibited an increase in the activated ERK1/2 pathway. As well, pro-fibrogenic cytokines increased the promoter activity of type I collagen, and this activity decreased with cells that were co-transfected with dominant negative plasmids of ERK1 and 2. RT-PCR confirmed that collagen transcript levels decreased when cells were transfected with dn ERK1 and 2 and then further stimulated with IL-4 and IL-13. These results were also mirrored with collagen secretion assays. In addition, we studied the role for transcription factor Elk-1 known to be activated via the ERK pathway. Dominant negative Elk-1 showed inhibition of collagen promoter activity in fibroblasts transfected with full collagen type I promoter or two fragments which contain the Elk-1 binding site. Our results suggest that the modulation of collagen gene expression may occur via the ERK pathway and is mediated by Elk-1.

Keywords:

• Collagen
• ERK
• Elk-1
• human fibroblasts
• fibrosis
• fibrogenic cytokines