The physiological expression of inducible nitric oxide synthase (iNOS) in the human colon

P J Roberts¹, G P Riley², K Morgan³, R Miller¹, J O Hunter¹, S J Middleton¹

¹ Unit E7, Box 201A, Department of Gastroenterology, Addenbrookes Hospital, CB2 2QQ Cambridge, UK
² Rheumatology Research Unit, Addenbrookes Hospital
³ Department of Biochemistry, University of Cambridge, Cambridge, UK

Correspondence to:
Dr Middleton stephen.middleton{at}msexc.addenbrookes.anglox.nhs.uk

Background—Inducible nitric oxide synthase (iNOS) is expressed in the colonic epithelium in both inflammatory bowel disease and colorectal cancer. Nitric oxide (NO), the product of this enzyme, has been implicated in the pathogenesis of both conditions. However, there are conflicting data on whether iNOS is expressed in the normal, uninflamed human colon.

Aims—To evaluate the expression of iNOS in histologically normal, non-inflamed human colonic mucosa.

Patients/Methods—Reverse transcription polymerase chain reaction (RT-PCR), immunoblotting, and immunohistochemistry were used to investigate the expression of iNOS in 17 histologically normal specimens obtained at colectomy performed for colorectal neoplasia. In addition, 16 endoscopic mucosal biopsies, taken from normal individuals, were also evaluated. Eleven surgical specimens and 16 endoscopic biopsies from patients with refractory ulcerative colitis were used as inflammatory controls.

Results—All types of specimens expressed iNOS mRNA. Immunoblotting revealed a protein of approximately 130 kDa consistent with iNOS in mucosal extracts of 77% of normal individuals, and 85% of diseased controls. Immunolabelling localised this protein to the surface epithelium in most of the normal specimens and also to the crypt epithelium and inflammatory cells in the diseased controls.

Conclusions—These findings provide evidence that iNOS is often expressed in the surface epithelium of non-inflamed human colon, suggesting that it is induced by local luminal factors, such as bacterial lipopolysaccharide (endotoxin). The resultant NO produced at this site might act as an oxidative barrier, reducing bacterial translocation and providing a means of defence against pathogenic microorganisms.

Key Words: inducible nitric oxide synthase • ulcerative colitis • nitric oxide • colorectal carcinoma

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