Stimulation of the extracellular matrix production in dermal fibroblasts by velvet antler extract

Abstract

Background: Fibroblasts produce many components of the extracellular matrix (ECM) and so they contribute to the maintenance of connective tissue integrity.

Objective: The aim of this study is to evaluate the effect of velvet antler extract (VAE) on the ECM production of dermal fibroblasts cultured in vitro.

Methods: Primary cultured human dermal fibroblasts were treated with VAE, and then the ECM production was determined by RT-PCR, ELISA and Western blot analysis. Furthermore, the change of gene expression according to VAE treatment was evaluated by cDNA microarray.

Results: VAE accelerated the growth of fibroblasts in a dose-dependent manner. VAE increased the production of several ECM components, including type 1 collagen, fibronectin and elastin. In line with these results, the phosphorylations of p42/44 ERK and p38 mitogen-activated protein kinase were markedly increased by VAE, suggesting that the enhancement of ECM production may be linked to the activation of intracellular signaling cascades. VAE also significantly increased cell migration on an in vitro scratch wound test. In cDNA microarray, many genes related with connective tissue integrity were identified to be up-regulated by VAE.

Conclusion: These results suggest that VAE has a potential to stimulate ECM production, and VAE may be applicable for maintaining the skin's texture.

Keywords: Extracellular matrix; Fibroblast; Velvet antler extract; cDNA microarray.

Fig. 1

Effect of velvet antler extract (VAE) on the growth of dermal fibroblasts. The cells were treated with VAE at the indicated concentrations for 2 days in the presence of [³H]thymidine. The radioactivity was measured by a liquid scintillation counter. The results are shown as percentage of the control±standard deviation (SD) (^*p<0.05 vs. control).

Fig. 2

Effect of velvet antler extract (VAE) on the extracellular matrix (ECM) production of dermal fibroblasts. The cells were treated with VAE at the indicated concentrations for 2 days. The conditioned medium was collected, and the secreted procollagen type 1 (A) and fibronectin (B) were measured by ELISA. The results are shown as percentage of the control±standard deviation (SD) (^*p<0.05 vs. control). (C) The cellular proteins were harvested and the protein levels for collagen type 1 α1 and elastin were verified by Western blot analysis.

Fig. 3

Effect of velvet antler extract (VAE) on intracellular signaling pathway. The cells were treated with VAE for the indicated time points. The cellular proteins were prepared and the phosphorylations of ERK and p38 MAPK were determined by Western blot analysis.

Fig. 4

Effect of velvet antler extract (VAE) on cell migration. Confluent monolayers of dermal fibroblasts were wounded using a pipette tip, and the monolayer cells were treated with VAE at the indicated concentrations. The areas of cell-free wounds were photographed after 24 h treatment.

Fig. 5

Dermal fibroblasts were treated with velvet antler extract (VAE) at the indicated concentrations for 2 days. The levels of expression of selected genes were verified by RT-PCR.