Immunomodulatory effects of inhaled carbon monoxide on rat syngeneic small bowel graft motility.
BACKGROUND
Intestinal transplantation provokes an intense inflammatory response within the graft muscularis that causes intestinal ileus. We hypothesised that endogenously produced anti-inflammatory substances could be utilised as novel therapeutics. Therefore, we tested the protective effects of inhaled carbon monoxide (CO) and an endogenous haeme oxygenase 1 (HO-1) anti-inflammatory mediator on transplant induced inflammatory responses and intestinal ileus in the rat.


METHODS
Gastrointestinal transit of non-absorbable FITC labelled dextran and in vitro jejunal circular muscle contractions were measured in controls and syngeneic orthotopic transplanted animals with and without CO inhalation (250 ppm for 25 hours). Inflammatory mRNAs for interleukin (IL)-6, IL-1beta, tumour necrosis factor alpha (TNF-alpha), intercellular adhesion molecule 1 (ICAM-1), inducible nitric oxide (iNOS), cyclooxygenase 2 (COX-2), and IL-10 were quantified by real time reverse transcriptase-polymerase chain reaction and HO-1 by northern blot. Histochemical stains characterised neutrophil infiltration and enterocyte apoptosis.


RESULTS
Transplantation delayed transit and suppressed jejunal circular muscle contractility. Transplantation induced dysmotility was significantly improved by CO inhalation. Transplantation initiated a significant upregulation in IL-6, IL-1beta, TNF-alpha, ICAM-1, iNOS, COX-2, and HO-1 mRNAs with the graft muscularis. CO inhalation significantly decreased expression of IL-6, IL-1beta, iNOS, and COX-2 mRNAs. CO also significantly decreased serum nitrite levels (iNOS activity).


CONCLUSIONS
CO inhalation significantly improved post-transplant motility and attenuated the inflammatory cytokine milieu in the syngeneic rat transplant model. Thus clinically providing CO, the end product of the anti-inflammatory HO-1 pathway, may prove to be an effective therapeutic adjunct for clinical small bowel transplantation.
Nakao
A
A
Thomas E Starzl Transplantation Institute, University of Pittsburgh, Pittsburgh, PA 15261, USA.
Moore
B A
BA
Murase
N
N
Liu
F
F
Zuckerbraun
B S
BS
Bach
F H
FH
Choi
A M K
AM
Nalesnik
M A
MA
Otterbein
L E
LE
Bauer
A J
AJ
D000818:Animals; D018723:Bethanechol / Q000494:pharmacology; D015152:Blotting, Northern; D002248:Carbon Monoxide / Q000008:administration & dosage*; D051546:Cyclooxygenase 2; D016207:Cytokines / Q000378:metabolism; D005769:Gastrointestinal Motility / Q000187:drug effects / Q000276:immunology* / Q000502:physiology; D005772:Gastrointestinal Transit / Q000276:immunology; D006419:Heme Oxygenase (Decyclizing) / Q000378:metabolism; D051547:Heme Oxygenase-1; D007249:Inflammation / Q000209:etiology / Q000503:physiopathology; D018799:Intercellular Adhesion Molecule-1 / Q000378:metabolism; D007375:Interleukin-1 / Q000378:metabolism; D016753:Interleukin-10 / Q000378:metabolism; D015850:Interleukin-6 / Q000378:metabolism; D007421:Intestine, Small / Q000276:immunology / Q000502:physiology / Q000637:transplantation*; D007527:Isoenzymes / Q000378:metabolism; D008297:Male; D009119:Muscle Contraction / Q000187:drug effects; D019001:Nitric Oxide Synthase / Q000378:metabolism; D011451:Prostaglandin-Endoperoxide Synthases / Q000378:metabolism; D051381:Rats; D017207:Rats, Sprague-Dawley; D020133:Reverse Transcriptase Polymerase Chain Reaction; D014409:Tumor Necrosis Factor-alpha / Q000378:metabolism
D016207:Cytokines; D007375:Interleukin-1; D015850:Interleukin-6; D007527:Isoenzymes; D014409:Tumor Necrosis Factor-alpha; D018723:Bethanechol; D018799:Intercellular Adhesion Molecule-1; D016753:Interleukin-10; D002248:Carbon Monoxide; D019001:Nitric Oxide Synthase; D006419:Heme Oxygenase (Decyclizing); D051547:Heme Oxygenase-1; D051546:Cyclooxygenase 2; D011451:Prostaglandin-Endoperoxide Synthases