Antibiotic treatment (Abx) of rats with anastomosis plus devascularization (Anast + Dvasc) attenuated MMP9 activation, and expression of HIF1-α (hypoxia inducible factor), iNOS (inducible nitric oxide) and MPO (myeloperoxidase). (A) Western blot analyses of rat tissues subjected to devascularization (Dvasc), anastomosis (Anast) or Anast + Dvasc, in the presence or absence of direct topical application of ciprofloxacin, metronidazole, and neomycin (topical Abx) via enema. Experiments were performed on 3 rats per group; displayed immunoblot is representative of all results. (B) The evaluation of band intensities using Image J software demonstrated the abundance of the active form of MMP9 in the rat group Anast + Dvasc, and loss of active MMP9 after topical application of antibiotics. n=3, p<0.01, Student t-test. (C) Zymography analysis confirmed the abundance of the active form of MMP9 in the Anast + Dvasc rat group, and loss of the active form of MMP9 by topical application of antibiotics.