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ObjectiveObserving the effect of subcutaneous and local administration of Pseudomonas aeruginosa preparation on the intestinal anastomotic healingin rat, and compared to 5-Fu, further investigate the mechanism for its experimental basis for clinical application.MethodsSixty-four Sprague-dawley (SD) rats were randomly divided into four groups with 16 rats for each. Group B and C received 0.5 ml of pseudomonas aeruginosa preparation subcutaneous injection qod from 3 days before to 14 days after the surgery. In addition, Group C received 0.5 ml of pseudomonas aeruginosa preparation local anastomotic spray in surgery. Group A and D received 0.5 ml of saline subcutaneous injection qod from 3 days before. In addition, Group D received 5-Fu (20 mg/kg) as intraperitoneal chemotherapy. Observed rats in each group after the general situation of those who died a clear cause of death; rats in each group after 7d and 14d, died after dislocating laparotomy, to determine the location anastomosis, anastomotic release adhesions with the surrounding tissue, Anastomosis as the center to cut about 1cm of the colon, and so divided into two parts, with 3% glutaraldehyde fixative organization pre-fixed in 1% osmium tetroxide fixative re-fixation, acetone dehydration series, embedded epoxy EP0N812, semi-thin section, light microscope positioning, Relehert thin slicer sections uranyl acetate and lead citrate double staining, JEM-120 transmission electron microscope observation of submucosal lymphatic endothelial cell morphology and ultrastructure. The other part fixed in 10% neutral paraformaldehyde, paraffin embedded sections were performed HE staining, Masson staining and immunohistochemical staining, light microscopy anastomotic healing, anastomotic inflammatory cell infiltration, detection of HPC, Expression of VEGF and PDGF-BB. SPSS13.0 the data using statistical software for statistical analysis.Results1、General Rats in each group recovered activity after 6h, 12h recovery diet. There was no abdominal wall of rats after wound infection, each group were party of deaths, replace them on time.2、After 7 days, B and C groups anastomotic inflammatory cell infiltration, collagen deposition, HPC, VEGF and PDGF-BB expression were significantly higher than that of A and D group, D group of the lowest indicators.3、Anastomotic tissue was observed under transmission electron microscopy. In the B and C group anastomotic tissue, goblet epithelial cells , submucosa of neutrophils, T cells, macrophages and other cells significantly increased, and a large, chromatin density, darker staining, mitochondrial and other organelles increased; the corresponding group D cells and mucosal epithelial cells under the less nuclear small, lighter staining, mitochondria and other organelles are also small.ConclusionsPA-MSHA can activate the immune system in rats, increased macrophages and other immune cell activity, witch can promote cell growth factor secretion and epithelial cell migration and proliferation, and promote the healing of intestinal anastomosis.Its principle is possible e to related with PA-MSHA activating the Toll-like receptorm.
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