Studies on Mustard-Stimulated Proteases and Inhibitors in Human Epidermal Keratinocytes (HEK): Development of Antivesicant Drugs

摘要

In the cultured HEK model, we observed that mustard stimulates protease activity, and the epidermaldermal attachment protein laminin-5 is a substrate. Addition of serine protease inhibitors (50 M ICD 2812 or 1 mM phenyl methyl sulphonyl fluoride (PMSF)), the metalloprotease inhibitor 1, 10- phenanthroline (1 mM), or the general caspase inhibitor Z-VAD-FMK (benzyl oxycarbonyl-Val-Ala-Asp (o-methyl-fluromethylketone), 10 M) to cells prior to mustard decreased the protease bands (zymography) and laminin-5 degradation (Western blotting, immuno-fluorescence). In conclusion, our results indicate that (a) mustard stimulates multiple proteases in the skin, and (b) protease inhibitors are prospective vesicant countermeasures.