| OBJECTIVE The polypeptide (PCF, Mr:800-l000) was isolated from Chlamys farreri using enzyme engineering technique. We studied the percutaneous absorption of PCF and the scavenge effects on free radicals in vitro. METHODS The concentrations of PCF filtered from the skin were detected with HPLC method on percutaneous absorption equipment in vitro. We first calculated the piled up percutaneous ratio of per area of PCF, then the permeating coefficient and the percutaneous absorption of PCF in different concentrations, pH values, with or without absorption promoter and different status of skin. Using chemiluminescence and electron spin resonance (ESR) techniques, we examined the scavenging properties of PCF on superoxide anions (O2 -), hydroxyl radical (OH-), peroxynitrite (ONOO-) and the inhibiting capacities of it on peroxidation of linoleic acid. RESULTS The results showed that the permeating coefficient of PCF increased obviously following the concentration increasing. At pH=6.0, with the absorption promoter Azone (2%), the permeating coefficient of PCF got to maximum respectively. PCF could scavenge oxygen free radicals including superoxide anions (O2 -) (EC5o=0.3mg/ml), hydroxyl radical (OH-) generated from the reaction systems (EC5o=0.2 g/ml) and effectively inhibit the oxidative activity of ONOO" (EC50=0.2mg/ml). The result of ESR indicates that PCF could inhibit the lipid peroxidation of linoleic acid notably. When the concentration of PCF was l.25mg.ml-1, the inhibitory ratio of PCF on the lipid peroxidation of linoleic acid was 43%. It is difficult to find another antioxidant which can scavenge so effectively both ONOO" andoxygen free radicals simultaneously. CONCLUTION PCF could hardly filtered and its percutaneous absorption was influenced by its concentrations, pH value, absorption promoter and the status of skin. PCF could scavenge free radicals and inhibit the peroxidation of linoleic acid in vitro. It may be a potential antioxidant from the marine products. |
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