title: Searching for the molecular benchmark of physiological intestinal anastomotic healing in rats: an experimental study.
abstract: OBJECTIVE
  This investigation focuses on the physiological characteristics of gene transcription of intestinal tissue following anastomosis formation.
  
  
  METHODS
  In eight rats, end-to-end ileo-ileal anastomoses were performed (n = 2/group). The healthy intestinal tissue resected for this operation was used as a control. On days 0, 2, 4 and 8, 10-mm perianastomotic segments were resected. Control and perianastomotic segments were examined with an Affymetrix microarray chip to assess changes in gene regulation. Microarray findings were validated using real-time PCR for selected genes. In addition to screening global gene expression, we identified genes intensely regulated during healing and also subjected our data sets to an overrepresentation analysis using the Gene Ontology (GO) and Kyoto Encyclopedia for Genes and Genomes (KEGG).
  
  
  RESULTS
  Compared to the control group, we observed that the number of differentially regulated genes peaked on day 2 with a total of 2,238 genes, decreasing by day 4 to 1,687 genes and to 1,407 genes by day 8. PCR validation for matrix metalloproteinases-3 and -13 showed not only identical transcription patterns but also analogous regulation intensity. When setting the cutoff of upregulation at 10-fold to identify genes likely to be relevant, the total gene count was significantly lower with 55, 45 and 37 genes on days 2, 4 and 8, respectively. A total of 947 GO subcategories were significantly overrepresented during anastomotic healing. Furthermore, 23 overrepresented KEGG pathways were identified.
  
  
  CONCLUSIONS
  This study is the first of its kind that focuses explicitly on gene transcription during intestinal anastomotic healing under standardized conditions. Our work sets a foundation for further studies toward a more profound understanding of the physiology of anastomotic healing.
authors.affiliation: Department of General and Visceral Surgery, University Hospital Freiburg, Freiburg, Germany.
authors.forename: Gabriel J
authors.initials: GJ
authors.lastname: Seifert
authors.forename: Michael
authors.initials: M
authors.lastname: Seifert
authors.forename: Birte
authors.initials: B
authors.lastname: Kulemann
authors.forename: Philipp A
authors.initials: PA
authors.lastname: Holzner
authors.forename: Torben
authors.initials: T
authors.lastname: Glatz
authors.forename: Sylvia
authors.initials: S
authors.lastname: Timme
authors.forename: Olivia
authors.initials: O
authors.lastname: Sick
authors.forename: Jens
authors.initials: J
authors.lastname: Höppner
authors.forename: Ulrich T
authors.initials: UT
authors.lastname: Hopt
authors.forename: Goran
authors.initials: G
authors.lastname: Marjanovic
mesh_terms: D000714:Anastomosis, Surgical; D000818:Animals; D020869:Gene Expression Profiling; D063990:Gene Ontology; D007413:Intestinal Mucosa / Q000378:metabolism*; D007422:Intestines / Q000601:surgery*; D008297:Male; D020411:Oligonucleotide Array Sequence Analysis; D016133:Polymerase Chain Reaction; D011897:Random Allocation; D017208:Rats, Wistar; D014945:Wound Healing / Q000502:physiology*