Different expression of protein in the supernatant of heat injured keratinocytes

Objective To compare the difference of protein expression in the supernatant of heat injured keratinocytes ( KC) and normal KC. Methods A model of heat injured KC was produced in vitro. The supernatant of normal KC and heat injured KC was collected after culture for 12 hours, and was ultrafiltered and lyophilized to get the protein. The protein sample was separated by immobilized pH gradient based two dimensional gel electrophoresis (2-DE). The gel was stained and the different expression of protein was analyzed using ImageMaster 2D analysis software. Results ①Average protein spots were 1 898 ±113, 1 877 ±97 in the supernatant of normal and heat injured KC and 1118 protein spots could be used for statistical analysis. ②Statistical result showed that 26 protein spots were significantly different between the two groups. 16 protein spots were higher in the supernatant of normal KC and then 10 protein spots were lower in the normal group. @16 protein spots, which included 10 kinds of proteins, were identified successfully as different spots. Lower expression proteins were alpha-enolase, actin cytoplasmic2, peroxiredoxin-4, phosphoglycerate mutase 1, G protein-regulated inducer of neurite outgrowth 1 in the supernatant of heat injured KC. Higher expression proteins in heat KC were purine nucleoside phosphorylase, tumor necrosis factor ligand superfamily memberl0, proteasome subunit alpha type-7, UDP-glucose 6-dehydrogenase in the supernatant of heat injuryed KC. Conclusions The result indicated that there are some significant different expression proteins in the supernatant of normal KC and heat injured KC.These findings provide new data for screening major molecules of tissue repair and finding the mechanism of wound repair. Key words: Keratinocyte; Electrophoresis,gel,two-dimensional; Proteome