title: Toll like receptors gene expression of human keratinocytes cultured of severe burn injury.
abstract: OBJECTIVE
  To evaluate the expression profile of genes related to Toll Like Receptors (TLR) pathways of human Primary Epidermal keratinocytes of patients with severe burns.
  
  
  METHODS
  After obtaining viable fragments of skin with and without burning, culture hKEP was initiated by the enzymatic method using Dispase (Sigma-Aldrich). These cells were treated with Trizol(r) (Life Technologies) for extraction of total RNA. This was quantified and analyzed for purity for obtaining cDNA for the analysis of gene expression using specific TLR pathways PCR Arrays plates (SA Biosciences).
  
  
  RESULTS
  After the analysis of gene expression we found that 21% of these genes were differentially expressed, of which 100% were repressed or hyporegulated. Among these, the following genes (fold decrease): HSPA1A (-58), HRAS (-36), MAP2K3 (-23), TOLLIP (-23), RELA (-18), FOS (-16), and TLR1 (-6.0).
  
  
  CONCLUSIONS
  This study contributes to the understanding of the molecular mechanisms related to TLR pathways and underlying wound infection caused by the burn. Furthermore, it may provide new strategies to restore normal expression of these genes and thereby change the healing process and improve clinical outcome.
authors.affiliation: Department of Surgery, EPM, UNIFESP, Sao Paulo, SP, Brazil.
authors.forename: Sarita Mac
authors.initials: SM
authors.lastname: Cornick
authors.affiliation: Department of Surgery, EPM, UNIFESP, Sao Paulo, SP, Brazil.
authors.forename: Silvana Aparecida Alves Corrêa de
authors.initials: SA
authors.lastname: Noronha
authors.affiliation: Department of Surgery, EPM, UNIFESP, Sao Paulo, SP, Brazil.
authors.forename: Samuel Marcos Ribeiro de
authors.initials: SM
authors.lastname: Noronha
authors.affiliation: EPM, UNIFESP, Sao Paulo, SP, Brazil.
authors.forename: Marcus V B
authors.initials: MV
authors.lastname: Cezillo
authors.affiliation: UNIFESP, Brazil.
authors.forename: Lydia Masako
authors.initials: LM
authors.lastname: Ferreira
authors.affiliation: Department of Surgery, Paulista School of Medicine, Federal University of Sao Paulo, Brazil.
authors.forename: Alfredo
authors.initials: A
authors.lastname: Gragnani
mesh_terms: D000328:Adult; D002056:Burns / Q000235:genetics*; D002478:Cells, Cultured; D004817:Epidermis / Q000293:injuries / Q000378:metabolism; D005260:Female; D015870:Gene Expression; D006801:Humans; D015603:Keratinocytes / Q000378:metabolism*; D008297:Male; D016133:Polymerase Chain Reaction; D012313:RNA / Q000302:isolation & purification; D051193:Toll-Like Receptors / Q000235:genetics* / Q000378:metabolism; D014945:Wound Healing
chemical_list: D051193:Toll-Like Receptors; D012313:RNA