Expression of skin-derived antileukoproteinase (SKALP) in reconstructed human epidermis and its value as a marker for skin irritation.
For the investigation of the skin irritancy potential of chemicals in an in vitro model, it is necessary to have sensitive end-points that predict the effects on native human skin. Our aim was to investigate whether the induction of the proteinase inhibitor SKALP in reconstructed epidermis can be used as a marker. The influence of culture conditions and the effect of topical application of sodium lauryl sulfate and oleic acid on SKALP expression were evaluated using immunohistochemistry and Northern blotting. SKALP expression was induced by serum, epidermal growth factor and fibroblasts. In the presence of retinoic acid and 1,25-dihydroxyvitamin D3 SKALP expression was inhibited, whereas supplementation with ascorbic acid and a-tocopherol had no effect. Tape-stripping of excised skin and topical treatment with sodium lauryl sulfate induced SKALP protein expression. Application of sodium lauryl sulfate and oleic acid on reconstructed epidermis also induced SKALP at the protein level but no significant effects could be demonstrated at mRNA levels. In conclusion, SKALP expression, which was increased upon application of sodium lauryl sulfate and oleic acid, can be used as an in vitro end-point for skin irritancy, irrespective of the modifying effects of culture conditions.
Boelsma
E
E
Department of Dermatology, Leiden University Medical Center, The Netherlands.
Gibbs
S
S
Ponec
M
M
D015415:Biomarkers; D015152:Blotting, Northern; D017449:Dermatitis, Allergic Contact / Q000201:enzymology* / Q000473:pathology*; D005260:Female; D015971:Gene Expression Regulation, Enzymologic; D006801:Humans; D066298:In Vitro Techniques; D007509:Irritants / Q000009:adverse effects*; D019301:Oleic Acid / Q000009:adverse effects; D014176:Protein Biosynthesis; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins / Q000378:metabolism*; D015842:Serine Proteinase Inhibitors / Q000378:metabolism*; D012867:Skin / Q000201:enzymology*; D012967:Sodium Dodecyl Sulfate / Q000009:adverse effects; D014815:Vitamins / Q000009:adverse effects
D015415:Biomarkers; D007509:Irritants; D053491:Proteinase Inhibitory Proteins, Secretory; D011506:Proteins; D015842:Serine Proteinase Inhibitors; D014815:Vitamins; D019301:Oleic Acid; D012967:Sodium Dodecyl Sulfate