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. 1998 Oct;43(4):564–570. doi: 10.1136/gut.43.4.564

Increased nitric oxide activity in a rat model of acute pancreatitis

R Al-Mufti 1, R Williamson 1, R Mathie 1
PMCID: PMC1727286  PMID: 9824587

Abstract

Background—Overproduction of nitric oxide (NO) via induction of the inducible NO synthase (iNOS) is an important factor in the haemodynamic disturbances of several inflammatory states. 
Aims—To identify the role of NO in a caerulein induced model of acute pancreatitis in the rat. 
Methods—Arterial blood pressure and plasma NO metabolites were measured at zero and seven hours in adult male Wistar rats administered caerulein (n=10) or saline (n=10). Pancreatic activity of NOS (inducible and constitutive) was assayed biochemically. The pancreatic expression and cellular localisation of NOS and nitrotyrosine (a marker of peroxynitrite induced oxidative tissue damage) were characterised immunohistochemically. 
Results—Compared with controls at seven hours, the pancreatitis group displayed raised plasma NO metabolites (mean (SEM) 70.2(5.9) versus 22.7 (2.2) µmol/l, p<0.0001) and reduced mean arterial blood pressure (88.7 (4.6) versus 112.8 (4.1) mm Hg, p=0.008). There was notable iNOS activity in the pancreatitis group (3.1(0.34) versus 0.1 (0.01) pmol/mg protein/min, p<0.0001) with reduced constitutive NOS activity (0.62 (0.12) versus 0.96 (0.08) pmol/mg protein/min, p=0.031). The increased expression of iNOS was mainly localised within vascular smooth muscle cells (p=0.003 versus controls), with positive perivascular staining for nitrotyrosine (p=0.0012 versus controls). 
Conclusions—In this experimental model of acute pancreatitis, iNOS induction and oxidative tissue damage in the pancreas is associated with raised systemic NO and arterial hypotension. Excess production of NO arising from the inducible NO synthase may be an important factor in the systemic and local haemodynamic disturbances associated with acute pancreatitis. 



Keywords: acute pancreatitis; nitric oxide; inducible nitric oxide synthase; peroxynitrite; caerulein induced pancreatitis

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Figure 1 .

Figure 1

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(A) Microscopic section of the pancreas from the control group, with normal appearance of acini. (B) Microscopic section of the pancreas from the pancreatitis group, showing the features of acute oedematous pancreatitis notably interstitial oedema, intracytoplasmic vacuoles, and extensive infiltration of the section with inflammatory cells (original magnification ×100). 


Figure 2 .

Figure 2

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Mean (95% CI) arterial blood pressure (MABP) throughout the procedure in the control and the pancreatitis groups. p<0.0001 for the interaction between group and time using split plot repeated measures ANOVA. There was a significant difference between the control group and the pancreatitis group at six hours (p=0.046) and seven hours (p=0.008) only (using Bonferroni correction). 


Figure 3 .

Figure 3

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(A) Mean arterial blood pressure (MABP) measurement at the start and end of the procedure (zero and seven hours). (B) Plasma NO metabolites concentrations at the start and end of the procedure in control and pancreatitis groups. 


Figure 4 .

Figure 4

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Biochemical assay of NOS activity. (A) iNOS activity (p<0.0001); (B) cNOS activity (p=0.031). 


Figure 5 .

Figure 5

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(A) Section from the pancreatitis group with positive staining for iNOS of the vascular smooth muscle cells. (B) Section from one of the three rats from the pancreatitis group with positive staining of the endothelial cells with the anti-iNOS antibody. (C) Section from the control group with negative staining of the vascular smooth muscle and endothelial cells with the anti-iNOS antibody (original magnification ×200). 


Figure 6 .

Figure 6

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(A) Pancreatic section from the pancreatitis group showing positive staining with the antinitrotyrosine antibody. (B) Section from the control group with negative staining for the antinitrotyrosine antibody (original magnification ×200). 


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