The concentration-dependent effect of anethole on collagen, MMP-2 and GAG in human skin fibroblast cultures.

Purpose: In aging skin and some skin disorders, components of skin extracellular matrix (ECM) are disturbed and therefore research to find skin drugs is important. Evaluation of anethole impact on collagen, GAGs and MMP-2 in human skin fibroblasts was the aim of this study. Materials and methods: For collagen assay the Sircol dye, 5-[H-3] proline and real time-PCR were used. MMP-2 activity was detected by zymography. GAG concentration was determined using 1,9-dimethylmethylene blue (DMMB). Cell viability was assayed with MTT. Results: In cells treated with 1 and 10 mu M anethole, a significant increase in collagen synthesis was demonstrated. In contrast, collagen synthesis was significantly decreased in cells exposed to 100 mu M anethole. Similar alterations were found in collagen type I expression. The concentration of collagen secreted into the medium was higher only in cells exposed to 1 mu M anethole, while it was lower under the influence of higher compound concentrations. It may be due to the lack of pro-MMP-2 activation at 1 mu M and a significant increase in the level of MMP-2 at 10 and 100 mu M anethole. GAG concentration was reduced under the influence of 100 mu M anethole, whereas anethole at lower concentrations revealed the ability to prevent H2O2-induced GAG increase. No significant cytotoxicity of anethole to fibroblasts was noted. Conclusions: Our findings demonstrate the concentration-dependent action of anethole on the crucial components of ECM in cultured skin fibroblasts, which may be somewhat beneficial and may possibly be developed towards a therapeutic use in some skin disorders.