Nitrite from inflammatory cells--a cancer risk factor in ulcerative colitis?

Elevated levels of luminal nitrite and a lowered luminal pH were found in 77 percent of patients with acute ulcerative colitis. No luminal nitrite was found in healthy control subjects. Nitrites are a secretory product of activated macrophages and neutrophils of the lamina propria, whereas the lowered luminal pH is due to diminished bicarbonate formation by impaired colonocytes. A hypothesis is put forward that nitrites, lowered pH, and bacterial amines are conducive to formation of carcinogenic n-nitroso compounds, which reflect a cancer risk in patients with ulcerative colitis dependent on the type and extent of inflammatory cell activation as well as metabolic impairment of colonic epithelial cells.

Nitrite and nitrate levels in ileostomy effluent: effect of dietary change.

1. Nitrite and nitrate levels were measured in samples from ileostomy bags or stomal samples of thirty-one ileostomists (twenty-two ulcerative colitis, nine Crohn's disease), 14-16 h after ingestion of a conventional meal or a meal containing a high content of nitrite and nitrate. 2. Ileostomy samples were decolourized with barium chloride, sodium sulphate and charcoal. Nitrite was determined spectrophotometrically by the Griess reaction and nitrate determined as nitrite after reduction with nitrate reductase (EC 1.7.99.4) in the presence of sodium formate. The mean percentage recovery from twenty-six spiked samples was 101.9 (SE 3.5)% for nitrite and 82.9 (SE 3.3)% for nitrate. 3. Ileostomy bag samples were obtained in twenty-nine cases of which ten had measurable nitrite (median 0, range 0-20.7 nmol/g) on a conventional meal compared with twenty-three cases (median 7.2, range 0-31.1 nmol/g) on the test meal (P less than 0.01). Nitrate levels were measurable in sixteen (median 6.7, range 0-48.2 nmol/g) after a conventional meal compared with twenty-one (median 20.5, range 0-53.2 nmol/g) after the test meal (P less than 0.01). 4. Stomal fresh-catch samples were obtained in twenty-four cases: combined nitrate and nitrite was higher in eighteen, lower in four and unchanged in two subjects after the test meal (P less than 0.05). 5. The type of foodstuff ingested can significantly alter measurable levels of nitrite-nitrate in the distal ileum and is one factor determining nitrite-nitrate input into the proximal colon.

Role of nitrite and nitrate as a redox couple in the rat colon. Implications for diarrheal conditions.

Colonic levels of nitrite and circulating levels of nitrate are elevated in subjects with chronic diarrhea. The role of colonic epithelial cells in oxidation-reduction of nitrite and consequent control of nitrite and nitrate levels is unknown. Isolated rat colonocytes and isolated loops of colon were used to study oxidation and reduction of nitrate and nitrite. Colonocytes oxidized nitrite to nitrate at a rate of 162 +/- 33 nmol/min.g (dry wt) (n = 6) over 0-20 min, a value increased by addition of 5 mM glucose and 1 mM nicotinamide adenine dinucleotide (p = less than 0.01), but not altered by antibiotics. Nitrite oxidation correlated linearly with the dry weight of isolated colonocytes (r = 0.92), indicating a cellular reaction. Nitrite was absorbed at a rate of 0.28 nmol/min.cm2 (n = 9) from the colonic lumen, which reflected 74% absorption of the available nitrite. Of the absorbed nitrite, 14.3% reappeared in the colonic lumen as nitrate. Colonocytes may be important in oxidizing nitrite to nitrate, thus preventing excess nitrite from entering into the circulation and returning some nitrate to the colonic lumen for respiration by anaerobic bacteria. The interaction of nitrites and colonocytes deserves further study with regard to the oxidation-reduction of nitrite and to the development of colonic neoplasia in chronic diarrheal conditions.

Effect of luminal or circulating nitrite on colonic ion movement in the rat.

The disposition of intravenously or luminally administered nitrite across the colonic mucosa and its effect on ion movement into or from the colon was assessed in anesthetized Porton rats using the isolated colon instilled either with sodium chloride (120 mM) or sodium chloride (80 mM) with sodium butyrate (40 mM). Ionic changes in the colon after intravenous injection of 10 mumol NaNO2 were compared with those occurring after injection of 10 mumol NaCl. After intravenous administration of nitrite, both nitrite and nitrate appeared in the colonic instillate in a ratio of 1:1. Nitrite increased chloride absorption (110%) and bicarbonate production (20%) when 40 mM butyrate was included in the instillate. Net sodium absorption, measured in the whole colon, was unchanged. Intravenous nitrite had no effect on ionic movement in the absence of butyrate. When NaNO2 (100 microM) was included luminally with the sodium chloride-butyrate instillate, bicarbonate production rate increased (25%), but sodium and chloride absorption were unaffected. Nitrite concentration in the instillate decreased during the 40-min experimental period at a rate of 0.275 nmol X min-1 X cm-2, and nitrate appeared at a rate of 0.037 nmol X min-1 X cm-2. We conclude that nitrate stimulates bicarbonate production in the colon, probably by stimulating the oxidation of butyrate, the main source of CO2 generation by the colonic mucosa.

Specific metabolic effect of sodium nitrite on fat metabolism by mucosal cells of the colon.

The effect on the mucosa of sodium nitrite that enters the colon from the ileum or transmucosally from the circulation is unknown. Isolated colonic mucosal cells and Roux-en-Y colostomies were used to test whether high doses of sodium nitrite (5-40 mM) had any harmful histological or inhibitory metabolic actions on the mucosa. Luminal instillation of 40 mM NaNO2 (3 ml/24 hr) for 7-14 days produced no microscopic changes in the mucosa either of damage (ulceration, mucus cell depletion) or of new growth (dysplasia, neoplasia). Beta oxidation of bacterial fatty acids (n-butyrate) by colonic epithelial cells in rat and man was enhanced by 50% (P less than 0.001) with 10 mM NaNO2, while oxidation of glucose and amino acid (proline) was not affected. Sodium nitrite significantly depressed ketogenesis (P less than 0.001) by the colonic mucosa of rat and man. In conclusion, sodium nitrite in the presence of bacteria has no damaging effect on the colonic mucosa but causes selective stimulation of fatty acid oxidation in the colonic mucosa of rat and man.

Anion control of sodium absorption in the colon.

The regulatory functions of anions in colonic absorption of sodium are unknown. Absorption of sodium ions was assessed with chloride, butyrate, nitrite, sulphate and oxalate anions in segments of proximal/distal colon and in defunctioned colon. Efficiency of sodium absorption was related to availability of CO2 in mucosal cells: CO2 availability was enhanced (P less than 0.01) by sodium nitrite or diminished (P less than 0.01) in defunctioned colon. Sodium nitrite stimulated absorption of sodium in the distal colon where bicarbonate secretion predominated and n-butyrate stimulated absorption of sodium in the proximal colon where hydrogen ion secretion predominated. Sodium absorption was very significantly diminished (P less than 0.01) in defunctioned colon. Results indicate that sodium absorption in the colon is both a double anion exchange system as well as cation/anion co-transport. Anions act differently on sodium absorption along the length of the colon and prolonged lack of anions plays a part in sodium malabsorption of the defunctioned colon.

Detectable colonic nitrite levels in inflammatory bowel disease--mucosal or bacterial malfunction?

In the healthy colon, sodium nitrite stimulates mucosal metabolism of short-chain fatty acids and absorption of ions, both functions that are impaired in the mucosa of patients with ulcerative colitis (UC). To assess the role of nitrite in colonic inflammatory disease, sodium nitrite was measured in rectal dialysate of 49 subjects (18 controls, 23 UC and 8 other colitis). None of the control or quiescent UC patients had measurable levels of nitrite while 78% of patients with acute UC and 38% of patients with other colitis had measurable nitrite levels (acute UC vs. other colitis chi 2 = 5.555, p less than 0.02). Functional activity of the colonic mucosa, judged by bicarbonate output, was impaired in all subjects with measurable nitrite levels in UC. Detection of nitrite in acute colitis suggests impaired oxidation of nitrite to nitrate in the colonic mucosa or impaired luminal reduction of nitrite to NH4 by bacteria.

Tissue metabolism of methionine in sheep.

The rate of oxidation of the carboxyl and methyl carbons of [14C]methionine to CO2 by homogenates of liver, kidney cortex, pancreas, muscle and small intestinal mucosa was studied in two breeds of sheep (Merino and Poll Dorset Horn) at three ages (2 weeks, 3 months, 4 years). Sodium alpha-keto-gamma- methiolbutyrate (0 X 4 mM) stimulated production of CO2 from the carboxyl carbon of methionine, but not from the methyl carbon. Sodium pyruvate did not affect the recovery of CO2 from either carboxyl or methyl of methionine. Sodium formate (15 mM) suppressed the conversion of the methyl carbon of methionine to CO2 by liver and kidney homogenates to 4 and 50%, respectively, of control values, but did not affect the percentage of carboxyl carbon of methionine recovered in CO2 with either tissue. With addition of S-methyl-L-cysteine (40 mM) and 3- methylthiopropionate (10 mM) the percentage of methyl and carboxyl carbons recovered in CO2 was reduced to about 20% of control values in homogenates of both tissues. Activity per gram of tissue was higher in liver and kidney cortex than in pancreas, intestinal mucosa, or muscle, with no significant differences due to breed (Merino or Poll Dorset Horn) or sex (ewe, ram or wether) of sheep. Conversion of both the carboxyl and methyl carbons to CO2 by liver was significantly lower in 2-week-old lambs than in older animals (P less than 0.01). The activity of other tissues was not markedly affected by age. Results are discussed in relation to evidence of alternative pathways of methionine catabolism, and capacities of the tissues of the sheep to catabolize methionine by alternative pathways.

The effect of diet and of methionine loading on activity of enzymes in the transulfuration pathway in sheep.

In three experiments, activity of hepatic enzymes associated with metabolism of methionine through the transulfuration pathway were studied with respect to possible effects of diet and methionine infusion per abomasum. In experiment 1 no differences in methionine adenosyltransferase (MAT) or cystathionine lambda-lyase (CGL) were detected between lucerne and wheaten straw diets, or between effects of fasting for 48 h and 96 h after feeding lucrene chaff as opposed to fasting after feeding wheaten straw. Fasting for 96 h resulted in a trend toward increasing CGL and MAT specific activities on both diets. In experiment 2 MAT was depressed significantly by infusion of methionine at 1.4 g/day and to a greater extent by infusion at 4.2 g/day, whilst CGL was not significantly affected. In experiment 3 MAT specific activity decreased significantly in response to both levels of methionine supplementation. Betaine-homocysteine methyltransferase activity was increased by methionine infusion. CGL decreased in all treatments but there was a larger decrease in those animals receiving methionine infusion. No significant changes were observed in relation to other enzymes examined which included cystathionine beta-synthase and threonine dehydratase. These observations are consistent with the hypothesis that in sheep the increase in methionine in blood plasma which occurs when methionine is absorbed in increased amounts may be due to reduced entry into the transulfuration pathway because of a repression of MAT activity.