| AIM:Proteomics was used to screen the biomarkers for predicting skin irritation. The protein-protein interaction between differentially expressed proteins biomarkers was studied, which would be helpful for the mechanistic understanding of the skin irritation.Methods:Keratinocytes were exposed to representative acid/basic irritants for 4h, and continued to culture for another 0,2,4,8 and 16h in the normal culture medium, then cells were collected.2-DE/MALDI-TOF/TOF MS proteomics was used to analyze the protein expression. Proteins with altered expression were further validated by Western blot and quantitative PCR. Localization of proteins in cells was detected by immunofluorescence staining and the concentration of IL-lalpha in culture medium was determined by ELISA. RNAi was used to study of the mechanism of skin Irritation related to HSP27 and CFL-1.Results:(1) 20 protein spots with altered expression were identified in keratinocytes exposed to SDS by 2DE-based proteomics technology; (2) In keratinocytes exposed to acid/basic irritants, the expression of Heat shock protein 27 (HSP27) and Superoxide dismutase[Cu-Zn] (SOD1) were down-regulated and the expression of CFL-1 (Cofilin-1) was up-regulated; (3) HSP27, SOD1 and CFL1 may be used as potential markers for predicting skin irritation; (4) The translocation of HSP27 was observed in nucleus, which could help to illustrate the related mechanism; (5) the highest expression of IL-lalpha was detected at 0h and 8h after exposure; (6) The expression of CFL-1 was significantly reduced after interference of HSP27, but it was upregulated after esposure to SDS.Conclusion:HSP27, SOD1 and CFL1 might be used as potential biomarkers for predicting skin irritation; Acute irritation might alter the cytoskeleton of keratinocyte, and these changes may be related to the interaction of HSP27 and CFL-1. |
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