ARECA CATECHU L. EXTRACT 291 and plusmin. In particular, their active sites can accommodate unsaturated fatty acids and fatty acid derivatives. It was therefore postulated that CC-516, composed of fatty acids, many of them amino acids, polypeptides and flavonoids, could inhibit HLE and thus attenuate HLE-catalyzed extracellular matrix alterations during skin inflammatory disorders. Figure 2 shows the concentration-dependent inhibition of PPE and HLE by CC-516. The CC-516 at 10-500 pg/ml as the final concentration exhibited 37% to 98% inhi- bition, and IC5o values were 40.8 pg/ml (PPE) and 48.1 pg/ml (HLE), respectively. Figures 3-5 show the effects of the CC-516 on the enzyme activity in vivo. The elastase affects a reduction in the number of elastin fibers at the level of the enzyme deposit. The number of fibers increased when we drifted from the enzyme deposit point. CC-516 has insignificant inhibitory effects at the concentrations of 10 pg/ml and 50 pg/ml. On the other hand, it reduces strongly the elastase activity and thus increases the number of fibers at 100 pg/ml, as shown in Figure 5. This highly protective behavior towards elastin degradation contrasts with the properties of other elastase inhibitors (6). CC-516 could protect elastic fibers against elastase degradation in an ex vivo assay. Table III shows the effect of CC-516 on the cell proliferation of human fibroblasts. CC-516 increased cell proliferation by 85% at 10-4% concentration, compared to that of the 120 lOO -• 80 .--- 60 ,- 40 20 0 --e- HLE -El- PPE 0 1 O0 200 300 400 500 Concentration (IzO/m I) Figure 2. AntJ-eJastase activity of CC-516. Dose-response curves for the inhibitory effects on PPE (IC5o: 40.8 pg/ml) and HLE (IC5o: 48.1 pg/ml).

292 JOURNAL OF COSMETIC SCIENCE Figure 3. Skin section incubated without elastase Elastin is the brown-red fibers. Figure 4. Skin section incubated with I I•g/ml elastase. Note the total disappearance of elastin fibers. control, whereas the increase by ascorbic acid was 50%. Data shown in the Table III are the cell viabilities (%) compared to those of the control (100%). The incorporation of amino acids and other compounds in the presence of CC-516 was compared to incor- poration alongside ascorbic acid, a known collagen, for synthesis stimulus. Results of the CC-516 are better than those induced by vitamin C when used at its optimal concen- tration for the synthesis of collagen and the activation of the hydroxylation enzyme (39-42). CC-516 at 10-5% and 10-4% concentration increases the monolayer collagen synthesis by 20% and 50%, respectively. Data shown in Table IV are the synthesis ratio (%) compared to the control (100%). The skin model is a three-dimensional model, close to the living model, which allowed us to confirm the stimulating effect of CC-516 on the synthesis of proteins. At the 0.1% concentration, CC-516 stimulates collagen syn-