Proteomic investigation of human burn wounds by 2D-difference gel electrophoresis and mass spectrometry.

摘要

BACKGROUND: In humans, thermal cutaneous injury represents a serious traumatic event that induces a host of dynamic alterations. Unfortunately the molecular mechanisms that underlie these serious perturbations remain poorly understood. We applied a global analysis method to identify dynamically changing proteins within the burn environment, which could eventually become biomarkers or targets for treatment. MATERIALS AND METHODS: Protein extracts of normal/unwounded skin and burn wounds were assayed by 2D-difference gel electrophoresis (DIGE), a proteomic technology by which abundance levels of intact proteins (including isoforms) were simultaneously quantified from multiple samples with statistical confidence. Through unsupervised multivariate principal component analysis, protein expression patterns from individual samples were appropriately clustered into their correct temporal healing periods grouped into postburn periods of 1-3 days, 4-6 days, or 7-10 days after injury. Forty-six proteins were subsequently selected for identification by matrix-assisted laser desorption/ionization time-of-flight mass spectrometry. RESULTS: Proteins identified with differential temporal patterns of expression included predictable cytoskeletal proteins such as vimentin, and keratins 1, 5, 6, 16, and 17. Other candidate proteins with potential involvement in healing included heat shock protein 90, members of the serpin family (Serpin B1, SCCA1 and -2), haptoglobin, gelsolin, eIF4A1, IQGAP1, and translationally controlled tumor protein. CONCLUSIONS: We have used the combined technique, DIGE/mass spectrometry, to capture new insights into cutaneous responses to burn trauma and subsequent processes of early wound healing in humans. This pilot study provides a proteomic snapshot of temporal events that can be used to weave together the interconnected processes that define the response to serious cutaneous injury.