ORIGINAL ARTICLE

Fish Oil Reduces Gastric Acid Secretion

C. Riber, M. Wjdemann, T. Bisgaard, H. Ingels, J. F. Rehfeld & O. Olsen Dept. of Surgical Gastroenterology C and Dept. of Clinical Biochemistry, Rigshospitalet, University of Copenhagen, Copenhagen, and Dept. of Surgical Gastroenterology D, Glostrup Hospital, University of Copenhagen, Glostrup, Denmark

Riber C, Wjdemann M, Bisgaard T, Ingels H, Rehfeld JF, Olsen O. Fish oil reduces gastric acid secretion. Scand J Gastroenterol 1999;34:845848.
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Background: The aim of the present investigation was to study gastric acid secretion and release of gastrin, cholecystokinin (CCK), and secretin during intraduodenal perfusion of either sh oil or trioleate. Methods: Seven healthy volunteers were stimulated on two separate days in random order with intraduodenal perfusates of either sh oil or trioleate. Results: Intravenous infusion with gastrin-17 was used as a background stimulation in doses mimicking a postprandial situation (39.9 4.8 pmol/l sh oil and 43.6 3.8 pmol/l trioleate). Gastric acid secretion increased signicantly from a basal level of 0.7 0.1 meq/15 min to 4.0 0.6 meq/15 min (P ` 0.05) before perfusion of sh oil, which reduced gastric acid secretion to 1.9 0.4 meq/15 min (P ` 0.01). After termination of sh oil perfusion gastric acid secretion increased to preperfusion concentrations (P ` 0.01). Perfusion of trioleate did not inuence gastric acid secretion. Plasma concentrations of CCK rose signicantly during perfusion of sh oil (from 2.8 0.6 pmol/l to 4.4 0.7 pmol/l, P ` 0.01), whereas trioleate only tended to increase CCK concentrations. Plasma concentrations of secretin did not change during perfusion of sh oil; however, concentrations were signicantly lower during and after perfusion of trioleate (P ` 0.01). Conclusion: The present study shows that intraduodenal perfusion of sh oil is associated with a signicant reduction of the gastric acid secretion stimulated by gastrin in healthy humans. Key words: Cholecystokinin; sh oils; gastric acid; gastrin; secretin; trioleate Claus Riber, M.D., Dept. of General Surgery, Herning Hospital, Gl. Landevej 61, DK-7400 Herning, Denmark (fax: 45 9721 2673)

at from sh is rich in omega-3 fatty acids. Recent studies have suggested that dietary sh oil protects against myocardial infarction (1, 2) and enhances defence mechanisms against gastroduodenal mucosal injury (3, 4). Whether the mucosaprotective mechanism is due to an acid-suppressing effect is largely unknown. In the small intestine long-chain fatty acids like oleic acid are known to inhibit gastric acid secretion (57). In one study (4) 3 weeks of dietary sh oil supplementation failed to alter basal or pentagastrin-stimulated gastric acid secretion. Consequently, the aim of the present study was to investigate the effect of intraduodenal perfusion of sh oil and trioleate on gastric acid secretion stimulated by gastrin. Materials and Methods Seven healthy volunteers, three women and four men, aged 22 to 28 years, participated in the study. In random order each subject on different days received either sh oil or trioleate during stimulation of gastric acid secretion with gastrin. After an overnight fast a double-lumen tube (Anderson

tube no. 10) was positioned in the duodenum under uoroscopic control with the infusion site in the second part of the duodenum and the aspiration sites located 2025 cm further distally. A gastric aspiration tube was positioned with its tip in the antrum. Gastric and duodenal contents were aspirated by an intermittent-suction device (Egnell, Sweden) at 50 mm Hg and collected in 15-min samples. After a 30-min period of equilibrium, with the subject in the supine position, perfusion of the duodenum with isotonic saline was started at time 0 min at a rate of 10 ml/min, which was held constant throughout the study. Likewise, at 0 min non-sulphated gastrin-17 (40 pmol/kg/h) was administered by continuous intravenous infusion for 165 min. This dose results in plasma gastrin concentrations comparable to those after a meal. Seventy-ve min after the start of the gastrin-17 infusion the duodenum was perfused with one of the two fat perfusates for 60 min at a rate of 10 ml/min. Finally, the duodenum was again perfused with isotonic saline for the last 30 min of the study. Venous blood samples (2 10 ml) were taken every 15 min and immediately placed on crushed ice and centrifuged at 4 C. Each tube contained either 247.5 KIU of

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aprotinin and 6.9 IU of heparin/ml blood or 500 KIU of aprotinin/ml blood and ethylenediaminetetraacetic acid (EDTA). Plasma was stored at 20 C until radioimmunoassay. The volumes of aspirates were measured, and the concentration of Hwas determined by titration to pH 7 with an autotitrator (Radiometer, Copenhagen, Denmark). Osmolality was determined by freezing-point reduction and used as an index of duodenogastric reux. Validation studies have shown that the reduction in osmolality of gastric juice correlates well with the degree of reux calculated in accordance with Faber et al. (8).
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Changes in outputs and concentrations during the period of observation were evaluated by the Friedman non-parametric repeated-measures test and the Dunn multiple comparisons test for differences between periods. Paired non-parametric statisticsthe Wilcoxon signed-ranks testwas used for differences between study days. The level of signicance was P ` 0.05 (14). Ethics The study was approved by the Local Ethical Committee, and informed consent was obtained in all cases. Results Intravenous infusion of gastrin-17 increased plasma concentrations on both study days from a basal of 8.0 0.6 pmol/l sh oil and 9.4 1.2 pmol/l trioleate to a plateau of 39.9 4.8 pmol/l and 43.6 3.8 pmol/l, respectively (P ` 0.01) (Fig. 1, upper panel) with no differences between study days. Perfusion of sh oil and trioleate had no effect on plasma gastrin. Accordingly, gastric acid secretion increased signicantly from 0.7 0.1 meq/15 min to 4.0 0.6 meq/15 min (P ` 0.05) (Fig. 2, upper panel) before perfusion of sh oil, which reduced acid secretion to 1.9 0.4 meq/15 min (P ` 0.01). After termination of sh oil administration gastric secretion increased to preperfusion concentrations (P ` 0.01). Perfusion of trioleate did not inuence gastric acid secretion in any signicant way, and comparable rates were seen during basal and before and after perfusion on both days (Fig. 2, upper panel). In terms of gastric acidity, a similar pattern was observed (Fig. 2, lower panel). Gastrin-17 per se had no effect on plasma concentrations of CCK, whereas perfusion of sh oil signicantly increased CCK concentrations from 2.8 0.6 pmol/l to 4.4 0.7 pmol/l (P ` 0.01) (Fig. 1, middle panel). Replacing sh oil with saline promptly and signicantly decreased plasma CCK to 1.5 0.3 pmol/l (P ` 0.01). Trioleate perfusion tended to increase plasma CCK to above the basal level, but it only reached statistical signicance after termination of trioleate (P ` 0.01). Plasma concentrations of secretin did not change signicantly during perfusion of sh oil, whereas secretin concentrations were signicantly lower during and after perfusion of trioleate (P ` 0.01 for both periods) (Fig. 1, lower panel). The time course of gastric volume secretion during intraduodenal perfusion of either sh oil or trioleate did not differ from each other during gastrin-17 stimulation (data not shown). The osmolality within the time intervals was identical for both study days. It increased slightly, but not statistically, throughout the study (data not shown). Discussion The present study points to a reduction of gastrin-17-

Hormone analysis Plasma gastrin concentrations (sum of exogenously administered non-sulphated gastrin-17 plus endogenously produced gastrin) were measured as previously described, using antiserum 2604 (9, 10), which binds gastrin-34 and gastrin-17 with equimolar potency. The binding of CCK peptides is negligible. The detection limit of the assay was 1 pmol/l. Expressed as the coefcient of variation, intra-assay precision was between 5% and 10% throughout the working range of the assay. Interassay precision was 13%. Plasma CCK concentrations were measured using antiserum no. 92128, which binds the bioactive forms of CCK in plasma with equimolar potency. This new CCK radioimmunoassay is highly sensitive, with a detection limit `0.1 pmol/l. The intra-assay variation was always `10% at relevant concentrations (that is, below 7 pmol/l), whereas the interassay variation was `13% at concentrations up to 7 pmol/l. The antiserum binds only fully sulphated and carboxyamidated CCK peptides (11). Plasma secretin concentrations were measured after extraction of plasma with ethanol as described previously (12, 13), using the secretin-specic antiserum 55953. The detection limit of the assay was 0.8 pmol/l. Intra-assay and interassay variations were 6% and 14%, respectively, at a mean concentration of 6.6 pmol/l. Preparations of fat All fat perfusates contained 665 ml NaCl 7 ml Polysorbat-80 (Sigma Chemicals Co., St. Louis, Mo., USA) and 35 ml fat (sh oil or trioleate). Trioleate (35 ml; 1210 kJ) consisted of monounsaturated oleic acid with 18 carbon atoms (95%) (Sigma Chemicals Co.). Fish oil (35 ml; 1269 kJ) was polyunsaturated fatty acids (triglycerides) with mainly (80%) 2024 carbon atoms in the chains; 24 g omega-3 fatty acids (Pikasol, Lube, Hadsund, Denmark). Statistical analyses Results are given as means standard error of the mean. Data from the last two 15-min samples of each period were pooled and used for comparisons between test periods.
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Fig. 2. Gastric acid secretion (upper panel) and gastric acidity (lower panel) during intraduodenal perfusion of sh oil (dotted bar) and trioleate (dark grey bar) on a background intravenous stimulation with gastrin-17 in seven healthy volunteers (mean standard error of the mean). Asterisks (*P ` 0.05; **P ` 0.02; ***P ` 0.01) indicate signicant changes above basal, before, during, and after perfusion of oil.

Fig. 1. Plasma concentrations of gastrin (upper panel), cholecystokinin (CCK) (middle panel), and secretin (lower panel) during intraduodenal perfusion of sh oil (dotted bars) and trioleate (dark grey bar) on a background intravenous stimulation with gastrin-17 in seven healthy volunteers (mean standard error of the mean). Asterisks (***P ` 0.01) indicate signicant changes above basal, before, during, and after perfusion of oil. Double crosses (#P ` 0.05, ###P ` 0.01) indicate signicant differences between sh oil and trioleate perfusion.

stimulated gastric acid secretion by intraduodenal perfusion of sh oil, as indicated by the time course in Fig. 2. This seems to be an immediate effect of sh oil, as no long-term effect on acid suppression has been observed on either basal or pentagastrin-stimulated gastric acid secretory rates before

and after 3 weeks of sh oil treatement (4). Gastric acidity remained unchanged by trioleate perfusion, whereas sh oil reduced acidity signicantly (P ` 0.01) (Fig. 2, lower panel). The precise mechanism that mediates this feedback inhibition remains unknown, although an array of putative enterogastrones seems involved (7, 1518). Our study conrms the role of CCK, as the presence of fat in the duodenum is a potent releaser of CCK into the blood stream. A differential specicity of sh oil and trioleate on CCK release is indicated in the present study, inasmuch as trioleate has a longer effect on CCK release than sh oil, although not being correlated to the return of gastric acid secretion to preperfusion levels after termination of perfusion (Fig. 1, middle panel, and Fig. 2, upper panel). Whether larger doses of trioleate could have an effect remains to be investigated in a proper dose-response study. Nevertheless, sh oil does not differ from other longchain triglycerides with regard to plasma CCK release (17). In studies with monounsaturated oleic acid, secretin has been suggested as a possible candidate (7, 15). This could not be
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C. Riber et al. of the duodenal mucosa in humans. Eur J Clin Invest 1991; 21:2307. Konturek JW, Thor P, Maczka M, Stoll R, Domschke W, Konturek SJ. Role of cholecystokinin in the control of gastric emptying and secretory response to a fatty meal in normal subjects and duodenal ulcer patients. Scand J Gastroenterol 1994;29:58390. Grant HW, Palmer KR, Kelly RW, Wilson NH, Misiewicz JJ. Dietary linoleic acid, gastric acid, and prostaglandin secretion. Gastroenterology 1988;94:9559. Petersen F, Olsen O, Jepsen LV, Christiansen J. Fat and gastric acid secretion. Digestion 1992;52:436. Faber RG, Russel RCG, Royston CMS, Whiteld P, Hobsley M. Duodenal reux during insulin-stimulated secretion. Gut 1974; 15:8804. Rehfeld JF, Stadil F, Rubin B. Production and evaluation of antibodies for the radioimmunoassay of gastrin. Scand J Clin Lab Invest 1972;30:22132. Stadil F, Rehfeld JF. Determination of gastrin in serum. Scand J Gastroenterol 1973;8:10112. Rehfeld JF. Accurate measurement of cholecystokinin in plasma. Clin Chem 1998;44:9911001. Schaffalitzky de Muckadell OB, Fahrenkrug J. Radioimmunoassay of secretin in plasma. Scand J Clin Lab Invest 1977;37: 15562. Schaffalitzky de Muckadell OB, Fahrenkrug J, Nielsen J, Westphall I, Worning H. Meal-stimulated secretin release in man: effect of acid and bile. Scand J Gastroenterol 1981;16: 9818. Siegel S Non-parametic statistics for the behavioural sciences. New York: McGraw-Hill, Inc.; 1956. Rhee JC, Chang TM, Lee KY, Jo YH, Chey WY. Mechanism of oleic acid-induced inhibition on gastric acid secretion in rats. Am J Physiol 1991;260:G56470. Schaffalitzky de Muckadell OB, Olsen O, Cantor P, Magid E. Concentration of secretin and CCK in plasma and pancreaticobiliary secretion in response to intraduodenal acid and fat. Pancreas 1986;1:53643. Isaacs PET, Ladas S, Forgacs IC, Dowling RH, Ellam SV, Adrian TE, et al. Comparison of effects of ingested mediumand long-chain triglyceride on gallbladder volume and release of cholecystokinin and other gut peptides. Dig Dis Sci 1987;32: 4816. Layer P, Holst PP, Grandt D, Goebell H. Ileal release of GLP-1. Association with inhibition of gastric acid secretion in humans. Dig Dis Sci 1995;40:107482. Kroop HS, Long WB, Alavi A, Hansell JR. Effects of water and fat on gastric emptying of solid meals. Gastroenterology 1979;77:9971000.

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conrmed in the present study, in which secretin concentrations remained low during perfusion with both sh oil and trioleate. Interestingly, secretin was signicantly higher during and after sh oil than trioleate perfusion. This could be caused by a small and unintended greater loss of acid into the duodenum due to incomplete recovery during perfusion of sh oil, indicated by the higher acid secretion (Fig. 2, upper panel). After gastric emptying of 80% of a meal the pH of the gastric contents decreases from pH 7 to pH 3. The presence of fat in ingested foods slows down gastric emptying rates (19). The combination of slower gastric emptying and inhibition of acid secretion may prolong the postprandial period in which the pH is high. Therefore, our results of short-term perfusion of sh oil indicate that the suppression of acid secretion might be a possible mechanism of the fatty acid of the omega-3 type to protect the gastroduodenal mucosa. This remains to be elucidated in future studies including histology of the mucosa. In conclusion, the present study shows that intraduodenal perfusion of sh oil is associated with a signicant reduction of the gastric acid secretion stimulated by gastrin in healthy humans. Acknowledgement This study was supported by grants from The Danish Hospital Foundation for Medical Research, Region of Copenhagen, The Faroe Islands and Greenland. References
1. Bang HO, Dyerberg J. Fish consumption and mortality from coronary heart disease. N Engl J Med 1985;313:8223. 2. Leaf A, Weber PC. Cardiovascular effects of n-3 fatty acids. N Engl J Med 1988;318:54956. 3. Faust T, Redfern JS, Lee E, Feldmann M. Effects of sh oil on gastric mucosal 6-keto-PGF1a synthesis and ethanol-induced injury. Am J Physiol 1989;257:G913. 4. Schepp W, Peskar BM, Trautmann M, Stolte M, Hagenmuller F, Schusdziarra V, et al. Fish oil reduces ethanol-induced damage Received 1 March 1999 Accepted 31 May 1999

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