THE ANTI-WRINKLE AND ANTI-MELANOGENIC EFFECTS 279 protein after induction by TNF-α was determined by immunoblotting, p-coumaric acid was shown to be highly potent in the suppression of MMP-1 expression (Figure 5B). This result implies that p-coumaric acid has an inhibitory effect on MMP-1 protein expression and Figure 4. Ef fect of SSE and f SSE on the expression levels of MMP-1, -2, and -3 protein. HDF-N cells were treated with different doses of SSE and f SSE in the presence TNF-α (10 ng/ml) for 24 h. Then, equal amounts of cell lysate were subjected to immunoblotting (n = 3). (A) Representative image of immunoblots. (B) Densitomet- ric analysis of immunoblots. The expression level of protein is expressed relative to the level of β-actin. * and # indicate p 0.05 versus untreated control and TNF-α treated group, respectively. SSE 50% ethanol extract of S. bicolor L. stalk, f SSE A. oryzae NK- fermented form of SSE, MMP matrix metalloproteinase, TNF-α tumor ne- crosis factor-α.

JOURNAL OF COSMETIC SCIENCE 280 augments the suppressive effect of f SSE on MMP-1. Consequently, the enhanced activities of f SSE may be associated with increases in p-coumaric acid resulting from fermentation. However, we cannot exclude the possibility that other components may be involved in the inhibitory effect of f SSE, either alone or a combinatorial manner. Figure 5. Identi fi cation of p-coumaric acid by HPLC analysis and the expression level of MMP-1 protein. (A) The amounts of p-coumaric acid in SSE and f SSE were determined by HPLC (n = 3). Commercial p-coumaric acid was used as a reference material. (B) HDF-N cells were treated with different doses of p-coumaric acid in the presence TNF-α (10 ng/ml) for 24 h. Then, equal amounts of cell lysate were subjected to immunob- lotting (n = 3). The level of MMP-1protein is expressed relative to that of β-actin. * and # indicate p 0.05 versus untreated control and TNF-α treated groups, respectively. SSE 50% ethanol extract of S. bicolor L, MMP matrix metalloproteinase, TNF-α tumor necrosis factor-α.