Elevated expression of iNOS mRNA and protein in coeliac disease.
BACKGROUND
The role of nitric oxide synthase (NOS) in the pathophysiology of coeliac disease (CD) was investigated.


METHODS
We examined mRNA (reverse transcription multiplex polymerase chain reaction) and protein expression (Western blotting) of i,e and nNOS in enterocytes isolated from the duodenum of patients with untreated CD (n=22) and iron deficiency anaemia (IDA, n=22). Expression of IL1beta and TNFalpha, two pivotal "NOS-controlling" cytokines, was also studied.


RESULTS
Enterocytes from both patient groups were negative for eNOS and TNF(alpha) message but positive for n and iNOS. nNOS gene expression was not statistically different between groups (158.38+/-29.11% vs. 114.95+/-24.17%, IDA vs. CD, p=0.07, Mann-Whitney U). iNOS expression was higher in patients with CD when compared to patients with IDA (96.95+/-17.82% vs. 48.76+/-8.07%, p<0.006). Low levels of IL1beta mRNA (15.66+/-3.70%) were detected in nine samples-all of these samples were isolated from patients with CD representing a positive result in 40% of coeliac patients. In support of these observations, patients with CD expressed more iNOS protein than those with IDA (159.7+/-14.9% vs. 69.8+/-20%, p<0.05).


CONCLUSIONS
These results suggest that iNOS could be an important mediator in coeliac disease. Expression of this regulatory protein may be under the control of IL1beta.
Daniels
Ian
I
David Evans Medical Research Centre, City Hospital, Nottingham, NG5 1PB, UK. iandaniels25@hotmail.com
Cavill
Donna
D
Murray
Iain A
IA
Long
Richard G
RG
D000328:Adult; D000368:Aged; D018798:Anemia, Iron-Deficiency / Q000201:enzymology; D015153:Blotting, Western; D002446:Celiac Disease / Q000201:enzymology*; D004586:Electrophoresis; D020895:Enterocytes / Q000201:enzymology; D005260:Female; D006801:Humans; D007375:Interleukin-1 / Q000235:genetics; D008297:Male; D008875:Middle Aged; D019001:Nitric Oxide Synthase / Q000032:analysis / Q000235:genetics*; D052247:Nitric Oxide Synthase Type II; D016133:Polymerase Chain Reaction; D012333:RNA, Messenger / Q000032:analysis*
D007375:Interleukin-1; D012333:RNA, Messenger; C496319:NOS2 protein, human; D019001:Nitric Oxide Synthase; D052247:Nitric Oxide Synthase Type II