Effect of Bilateral Adrenalectomy on Gastric Acid and Pepsin Secretion from Gastric Fistulas and Heidenhain Pouches in Dogs

Vol. 52, No.3

GASTROENTEROLOGY

Copyright

© 1967 by The Williams

Printed in U.S.A.

& Wilkins Co:

EFFECT OF BILATERAL ADRENALECTOMY ON GASTRIC ACID-AND PEPSIN SECRETION FROM GASTRIC FISTULAS AND HEIDENHAIN POUCHES IN DOGS ALLAN R. COOKE, M.R.A.C.P., DAVID L. NAHRWOLD, M.D., AND MORTON I. GROSSMAN, M.D.

Veterans Administration Center and Departments of Medicine and Physiology, UCLA School of Medicine, Los Angeles, California

In previous studies, we have reported that bilateral adrenalectomy in dogs with Heidenhain pouches reduced the maximal acid output in response to histamine 1 but not to gastrin. 2 This is a report of the effect of bilateral adrenalectomy on innervated (gastric fistula) and denervated (Heidenhain pouch) gastric mucosa studied simultaneously.

Materials and Methods Four dogs were used. In 2 animals, a Thomas cannula" was put into the most dependent part of the stomach to form a gastric fistula (group 1). In the 2 other animals, a pouch of the oxyntic gland area (Heidenhain) was made. Three weeks later, a Thomas cannula was put into the most dependent part of the residual stJomach so that these animals (group 2) had both a gastric fistula (GF) and a Heidenhain pouch (HP). Tests were started approximately 3 weeks later. The animals were fasted 18 hr before each test. A polyethylene catheter (P.E. 50 Intramedic, Clay Adams, Inc., New York) was inserted into a hind leg vein, and an intravenous infusion of 0.15 M NaCI was maintained throughout each experiment by a peristaltic pump (Harvard Apparatus Company, Dover, Mass.) set to deliver 29 ml per hr. Gastric juice was collected from the GF and HP for two 15-min periods to obtain basal levels of secretion. At the Received September 6, 1966. Accepted October 27,1966. Address requests for reprints to: Morton I. Grossman, M.D., Building 114, Room 231, Veterans Administration Center, Los Angeles, California 90073. This work was supported in part by Grant AM 08354 from the United States Public Health Service. The authors are grateful to Ray Lichter for technical assistance.

end of this period, either gastrin or histamine was added to the saline to give the desired dosage. Doses were doubled every 60 min. During the 1st hr of each test, 1 liter of 0.15 M N aCI was given by subcutaneous infusion in order to maintain the dog in normal hydration throughout the experiment. Gastric juice was collected every 15 min, and the volume was recorded to the nearest 0.1 ml. The concentration of acid was determined by titration of a 0.2-ml sample of juice with 0.2 N NaOH to pH 7 using a glass electrode and an automatic titrator (Radiometer, Copenhagen). Pepsin activity of juice from the gastric fistula was measured by the method of Grossman and Marks using radioiodinated serum albumin as substrate.' Gastrin extracts were prepared from the mucosa of the pyloric gland area of hog stomach by the method of Gregory and Tracy carried only through the stage of fractionation with isopropanol." All the gastrin used was from the same batch. Doses of gastrin are expressed in terms of the wet weight of mucosa from which the extract was obtained. Histamine doses are recorded in terms of the weight of dihydrochloride salt. After control studies, each animal underwent bilateral adrenalectomy. The adrenalectomized animals were maintained on sodium chloride, approximately 4 g per day, plus desoxycorticosterone acetate (DOCA), 2.5 mg intramuscularly every 2nd day. The animals remained in good health. Careful attention was paid to the state of hydration, and, if any animal showed evidence of poor water or food intake, it was given 500 ml of 0.15 M N aCI subcutaneously daily until intake of food and water returned to normal. Studies were commenced on the 3rd postoperative day. Following the completion of studies on DOCA plus N aCI, the animals of group 2 (HP and GF) were given an intravenous infusion of hydrocortisone and then immediately started on hydrocortisone, 25 mg intramuscu-

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larly every 2nd day, and restudied. One animal of group 1 (GF) was tested for the effect on blood pressure of a continuous infusion of histamine at a high dose (6.0 mg per hr). The pressure was determined by auscultation using a mercury sphygmomanometer and a cuff wrapped around the hind leg.

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FIG. 1. Effect of bilateral adrenalectomy on acid output in gastric fistula dogs (group 1) in response to histamine and gastrin. The doses of both drugs are plotted on a logarithmic scale. Each point represents the mean acid output of the last two 15-min periods; each line, the mean of 3 experiments in each dog; vertical bars, SE. In dog 1 (A), gastrin studies were done on the 19th, 30th, and 76th days, and histamine studies on the 14th, 23rd, and 45th days after operation. In dog 2 (B), gastrin studies were done on the 19th, 25th, and 30th days, and histamine studies on the 14th, 23rd, and 27th days after operation.

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FIG. 2. Relation between time after adrenalectomy and the maximal acid output from gastric fistula and Heidenhain pouch in response to gastrin and histamine in dog 3. Each point is the maximal acid output on the day studied. The control studies are the mean maximal output of 3 experiments in 1 dog; vertical bars, SE.

put to both gastrin and histamine was markedly reduced. The mean maximal output with gastrin postadrenalectomy was 49% (fig. IA) and 42% (fig. IB) of control values. In response to histamine, the mean maximal acid output after adrenalectomy was 39% (fig. IA) and 49% (fig. IB) of control values. Both adrenalectomized animals collapsed at the highest dosage of histamine (6.0 mg per hr). We have described this in a previous communication.! Time course of gastric secretion following adrenalectomy in animals of group 2 (GF and HP). During control studies, the maximal acid output in response to histamine was generally equal to gastrin from the gastric fistula (figs. 2B and 3B) but greater than gastrin from the Heidenhain pouch (figs. 2A and 3A). Following adrenalectomy, the maximal acid output from the gastric fistula decreased in response to histamine and gastrin (figs. 2B and 3B) but only to histamine from the Heidenhain

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cortisone infusion was stopped, 25 mg of intramuscular hydrocortisone were, given. Studies 24 hr later showed that acid output from the GF and HP (histamine-stimulated) had returned to preadrenalectomy values. When tested on the 3rd day using gastrin, acid output from the GF and HP was approximately the same as before adrenalectomy (figs. 2 and 3). Effect of a large dose of histamine on blood pressure in the adrenalectomized animal. During the infusion of 1.0 mg per hr of histamine, the systolic pressure of the normal animal remained relatively stable; in the adrenalectomized animal, there was a drop of 20 mm Hg (fig. 6). However, with the commencement of 6.0 mg per hr of histamine, the adrenalectomized animal became extremely hypotensive over a period of 2 to 3 min (fig. 6). The animal collapsed and systolic blood 4

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pouch (figs. 2A and 3A). The maximal POST 3 acid output from the Heidenhain pouch due to gastrin stimulation (figs. 2A and 3A) was unaffected by adrenalectomy. The de2 c crease in acid secretion in the GF and the PR£-AORX E HP was apparent by the 5th to 6th day 10 and well established by the 10th day. From this day on, the acid secretion remained ~ !l relatively constant at this reduced value ~ 0+----,----.----.---, 10 20 40 (figs. 2 and 3). There was no apparent dif... BASAL 5 GASTRIN, g/hr ference in the time course between gastric ~ 4 B fistulas and Heidenhain pouches. IPepsin output from GF following adre:l o 3 nalectomy (groups 1 and 2). Following biz lateral adrenalectomy, pepsin output was Ul Q. not significantly different from control Ul 2 Q. responses. Gastrin- and histamine-stimulated pepsin outputs before and after adrenalectomy are presented in figure 4. Effect of hydrocortisone administration. On the 27th day after adrenalectomy, 0+----,----.----.---, BASAL 0.75 1.5 3.0 6.0 acute administration of hydrocortisone HISTAMINE 2HCI, mg/hr (100 mg initially, then 25 mg per hr intravenously for 6 hr) was tested on the 2 FIG. 4. Pepsin output from the gastric fistula animals with GF and HP. There was no in response to gastrin (A) and histamine (B) change in acid output, as compared to con- before and after adrenalectomy. Each line is the trol, in the GF or the HP (fig. 5). Im- mean of 12 experiments in 4 dogs; vertical lines, mediately after the intravenous hydro- SE.

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pressure could not be recorded, but it quickly recovered when histamine was stopped. The normal animal showed a drop of 20 mm Hg systolic and diastolic pressure but remained standing throughout the infusion.

Histamine dihydrochloride 0.75 mg Ihr Hydrocortisone 25mg/hr jv

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w ..: Discussion 100 ~ cl 0.: O~~__-r__~-''--THy~d~ro~~~rl~~O~Mr-~ Although there have been studies of the :i 2 3 4 5 6 7 effect of bilateral adrenalectomy on acid TIME IN HR secretion in the pylorus-ligated rat 6 - 14 and FIG. 5. Effect of an acute intravenous infusion pouch dogs,!' 15, 16 there have been only of hydrocortisone on acid output from gastric two studies in gastric fistula animals. In 1 fistula and Heidenhain pouch using background anesthetized cat, Corral-Saleta 17 found histamine stimulation. In the control situation, gastric secretion to histamine stimulation only histamine was infused. Mean of 2 experiwas reduced. In an excellent study by ments in 2 dogs. Bralow et aU 8 using chronic fistula rats, it was reported that basal acid and pepI Saline I Hist.2HCI I.Omg/hrlHisl.2HCI 6.0mg/hrl sin secretion were decreased exponentially 160 lLJ A---A}Systolic with time after operation. There have been 0:: A__ -A- __ ~ lt.~ ; :. no studies reported using gastric fistula ~o... fA ffl 120 dogs. 0:: i. " d' }[)'f" 'IDS OIIC 0' ' ~ a. II In this study, we have found that maxi- o 'I mal acid output in response to histamine g 80 ,:A ..J : Adrx animal collapsed, and gastrin was markedly reduced after III ,:~ infusion of Histamine stopped ---Adrx bilateral adrenalectomy in all 4 gastric 40 -Normal fistula animals (figs. 1 through 3). The reduction in acid output was time related o~---r--'.---.----.----.----30 60 90 120 150 up until about the 10th day; from then TIME IN MIN until the 76th day (fig. lA, gastrin), there FIG. 6. Effect on the blood pressure (systolic was no further decrease in acid secretion. This i~ in contrast to the findings of Bralow and diastolic) during the infusion of a high dose et al.;18 who reported that acid output de- of histamine in an adrenalectomized and a norcreased exponentially with time. There mal animal. Mean of three experiments in each are a number of differences in our study animal. and that of Bralow et al. Quite apart from a species difference, no attempt was made findings in HP (vagally denervated) dogs in the report of Bralow et al. to maintain (Cooke and Grossman, unpublished obnormal hydration, whereas we corrected servations). In the rat,6, 11, 13, 18 all reany obvious dehydration. Furthermore, ports (pylorus-ligated and chronic gastric only basal secretion was studied by Bralow fistula) have indicated a decreased pepsin et al., and thus it is not known whether the output following adrenalectomy. In the parietal cells were capable of responding Heidenhain pouch dog16 maintained on to an adequate stimulus. Although we DOCA, it was reported that bilateral only collected basal juice for 30 min, we adrenalectomy reduced basal pepsin secrenoted no tendency to develop basal ana- tion, but this conclusion was based on two cidity following adrenalectomy (fig. 1A experiments in 1 animal. In another reand B). port19 using Heidenhain pouch dogs, it Pepsin output in the GF (vagally in- was found that SU4885, an l1,B-hydroxylnervated) was unaffected by adrenalec- ase inhibitor, decreased acid secretion but tomy, even though acid output was de- had no effect on pepsin secretion; our findcreased (fig. 4). We have obtained similar ings are in keeping with this study. 200 .,.

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Hydrocortisone restored acid output in adrenalectomized GF and HP dogs (figs. 2 and 3), a finding that we have previously reported for HP dogs only.1 This is in agreement with other studies on the dog. 15 , 20, 21 Acute glucocorticoid or adrenocorticotropin administration has been shown to have either no effect on basal gastric acid secretion from pouches in dogs with intact adrenals 22-25 or only a slight effect. 26 In this study with adrenalectomized animals, hydrocortisone given for a period of 6 hr on a background of histamine stimulation was without effect on gastric acid output from the GF and the HP (fig. 5). In the Heidenhain pouch (vagally denervated gastric mucosa), adrenalectomy reduced the maximal response to histamine but not to gastrin, as we have previously reported. 2 Vagal denervation of the oxyntic glands (accomplished by transforming a Pavlov pouch into a Heidenhain pouch) depresses the maximal response to gastrin but does not alter the maximal response to histamine. 25 The present study shows that, when the maximal response to gastrin has been depressed by vagotomy, it is not further altered by adrenalectomy. By contrast, the maximal response to histamine is not depressed by vagal denervation, but it is decreased by adrenalectomy, which has about the same effect (about 50% depression) in the presence or absence of vagal innervation. We have previously reported that a large dose of histamine will cause collapse in adrenalectomized animals.! The mechanism is apparently cardiovascular, since marked hypotension occurred (fig. 6). Although Dolcini et al,28 have reported that bilateral adrenalectomy reduced gastric blood flow in the rat, it would be difficult to interpret the results of the present experiments on that basis alone. Summary

The effect on gastric secretion of bilateral adrenalectomy was studied in dogs with a vagally innervated gastric fistula and a vagally denervated Heidenhain pouch. Before adrenalectomy, maximal

Vol. 52, No.3

acid output from the gastric fistula was the same with stimulation by histamine or gastrin; bilateral adrenalectomy reduced maximal acid output to both of these stimulants by about 50%. Before adrenalectomy, maximal acid output from the Heidenhain pouch with stimulation by gastrin was about 50% of the value observed with histamine; bilateral adrenalectomy reduced the response to histamine by about 50% but did not change the response to gastrin, so that the maximal responses to these two stimulants were then about equal. Pepsin output from the gastric fistula in response to histamine or gastrin was not significantly altered by adrenalectomy. Hydrocortisone restored acid secretion to normal. In response to large doses of histamine, adrenalectomized animals collapsed. This collapse was due to severe hypotension. REFERENCES 1. Cooke, A. R, R M. Preshaw, and M. 1. Grossman. 1966. Effect of adrenalectomy and glucocorticoids on the secretion and absorption of hydrogen ion. Gastroenterology 50: 761-767. 2. Cooke, A. R, R M. Preshaw, and M. I. Grossman. 1965. Gastric mucosal hydrogen ion transfer and adrenocorticoids. Clin. Res. 13: 253. 3. Thomas, J. E. 1941. An improved cannula for gastric fistulas. Proc. Soc. Exp. BioI. Med. 46: 260-261. 4. Grossman, M. I., and I. N. Marks. 1960. Secretion of pepsinogen by the pyloric glands of the dog with some observations on the histology of the gastric mucosa. Gastroenterology 38: 343-352. 5. Gregory, R A., and H. J. Tracy. 1964. The constitution and properties of two gastrins extracted from hog antral mucosa. Gut 5: 103-114. 6. Tuerkischer, E., and E. Wertheimer. 1945. Adrenalectomy and gastric secretion. J. Endocr. 4: 143-151. 7. Madden, R J., and H. H. Ramsburg. 1951. Gastric secretion in the adrenalectomized rat. Endocrinology 49: 82-85. 8. Madden, R. J., and H. H. Ramsburg. 1951. Adrenalectomy in the Shay rat. Gastroenterology 18: 128-134. 9. Welbourn, R B., and C. F. Code. 1953. Effects

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