JOURNAL
OF SURGICAL
21, 357-358 (1976)
RESEARCH
A Simplified
Method
of Gastric JAMES
for Chemical Hypersecretion
Induction ’
B. WEEKS, M.D., GEORGE C. PETRIDIS, EDWARD R. WOODWARD, M.D., AND LESTER R. DRAGSTEDT, PH.D., M.D.”
M.D.,
Department of Surgery, College of Medicine, University of Florida, Gainesville, Florida 32610. Submitted for publication July 13, 1976
In 1942, Hay et al. [l] introduced a method of inducing gastric hypersecretion using intramuscular injections of crystalline histamine mixed with beeswax and mineral oil. This resulted in gradual histamine release with prolonged gastric hypersecretion in the experimental animal. This classic experiment established the hypothesis that peptic ulcer could be produced by an overproduction of acid gastric juice. The method used in these pioneer efforts is cumbersome, in both preparation and administration. We have, therefore, developed and tested an alternative method. With the advent of immunology, Freund’s adjuvant has become a commonly used medium for prolonged antigen exposure. While Freund’s complete adjuvant contains a protein moiety (Mycobacterium), the incomplete adjuvant is merely paraffin (mineral) oil [2] which can easily be emulsified with crystalline histamine dissolved in normal saline.
and all air is evacuated. An identical volume of Freund’s incomplete adjuvant is drawn into a second glass syringe and, again, all air is evacuated. The two syringes are connected by a double Luer-lok adapter with a small central aperture. The two solutions are emulsified by injecting back and forth between the two syringes for 5 min. A milky white viscous emulsion results. When emulsification is complete, a single drop placed in water will not show any dispersion. RESULTS
Multiple trials with various dosages and concentrations of histamine were tried in dogs in an attempt to produce gastric secretion for 24 hr without side effects. An intramuscular dosage of 10 mg/day in a concentration of 20 mg/ml was found to give the best gastric secretory response without side effects. Heidenhain pouch acid output, though tapering off with time, did last a full 24 hr with an average daily acid output of 90 METHODS mequiv. Table 1 demonstrates the volume Histamine was used in the dihydrochloride and milliequivalents of acid output at various form which is very hygroscopic and, times. Since acid output decreases with time therefore, must be desiccated immediately after injection, maximal stimulation can best prior to use. One gram of the desiccated be achieved by giving the injection apdihydrochloride contains 604 mg of histamine proximately 6 hr after feeding. Thus the base. The desired quantity is dissolved in a animal’s endogenous gastric secretory output volume of saline appropriate to make a solu- in response to the meal is followed by the tion twice the actual desired concentration. long-lasting exogenous stimulus of the This solution is drawn into a glass syringe emulsified histamine solution. The ulcerogenic potential for emulsified histamine was tested in two groups of 10 ‘This work has been supported by National Institutes dogs each. In Group I, resection of the of Health Grant No. AM-04178. ‘Deceased. gastric antrum was performed with re357 Copyright o 1976 by Academic Press, Inc. All rights of reproduction in any form reserved.
358
JOURNAL
OF SURGICAL
RESEARCH:
TABLE 1 Histamine-Stimulated Gastric Secretion’ Time W 2 4 6 8 10 12 14 16 18 20 22 24
5mgat 50 mg/ml 16.2 7.4 6.2 5.6 1.1 0.5
IOmgat 20 mg/ml 9.5 13.6 12.3 11.3 8.8 9.5 7.8 6.5 4.3 3.0 1.4 1.2
15mgat 20 mg/ml 12.6 14.3 14.5 12.2 10.7 9.5 6.2 5.0 2.0 I .4 1.3 0.8
“All values expressed in milliequivalents per 2-hr collection period. The 5 and IO-mg doses had no associated ill effects. The 15-mg dose was always accompanied by nausea and/or diarrhea.
construction using the Billroth II method with closure of the duodenal stump and endto-side gastrojejunostomy. In Group II, the antrum was resected and the duodenal stump was closed, but reconstruction was performed with a Roux-en-Y gastrojejunostomy. The animals were permitted a recovery period of 3 weeks, at which time food intake, nutritional state, and body weight were stabilized. The emulsified histimine injections were then begun. In Group I (gastrojejunostomy), marginal ulcers appeared in 3 to 31 days in 4 of the 10 animals (40%). In the Roux-en-Y group, as one would expect from the classic experiment of Mann
VOL. 21, NO. 5, NOVEMBER
1976
and Williamson [3], the incidence of marginal jejunal ulcer was 100%. Surviving animals were all sacrificed at the end of 31 days. SUMMARY
AND CONCLUSIONS
A method has been described for the emulsification of crystalline histamine in incomplete Freund’s adjuvant. Intramuscular injection results in slow release with strong stimulation of gastric secretion over a period of 18 to 24 hr. The ulcerogenic potential for this method was established in two groups of dogs. In dogs with antrectomy and Billroth I1 gastrojejunostomy, 40% developed a marginal ulcer with a dose of 10 mg of histamine base per day. In a comparable group of antrectomized dogs with reconstruction by Roux-en-Y gastrojejunostomy, diversion of alkaline duodenal fluids from the stoma increased the ulcerogenic potential of chemical gastric secretory stimulation and 100% of the animals developed marginal jejunal ulcer. REFERENCES Hay, L. J., Varco, R. L., Code, C. F., and Wangensteen, 0. H. The experimental production of gastric and duodenal ulcers in laboratory animals by the intramuscular injection of histamine in beeswax. Surg. Gynecol. Obsfet. 75:170, 1942. Freund, J. The effect of paraffin oil and mycobacteria on antibody formation and sensitization: A review. Amer. J. Clin. Pathol. 21:645, 195 1. Mann, F. C., Williamson, C. S. The experimental production of peptic ulcer. Ann. Surg. 77:409-422, 1923.
OF SURGICAL
21, 357-358 (1976)
RESEARCH
A Simplified
Method
of Gastric JAMES
for Chemical Hypersecretion
Induction ’
B. WEEKS, M.D., GEORGE C. PETRIDIS, EDWARD R. WOODWARD, M.D., AND LESTER R. DRAGSTEDT, PH.D., M.D.”
M.D.,
Department of Surgery, College of Medicine, University of Florida, Gainesville, Florida 32610. Submitted for publication July 13, 1976
In 1942, Hay et al. [l] introduced a method of inducing gastric hypersecretion using intramuscular injections of crystalline histamine mixed with beeswax and mineral oil. This resulted in gradual histamine release with prolonged gastric hypersecretion in the experimental animal. This classic experiment established the hypothesis that peptic ulcer could be produced by an overproduction of acid gastric juice. The method used in these pioneer efforts is cumbersome, in both preparation and administration. We have, therefore, developed and tested an alternative method. With the advent of immunology, Freund’s adjuvant has become a commonly used medium for prolonged antigen exposure. While Freund’s complete adjuvant contains a protein moiety (Mycobacterium), the incomplete adjuvant is merely paraffin (mineral) oil [2] which can easily be emulsified with crystalline histamine dissolved in normal saline.
and all air is evacuated. An identical volume of Freund’s incomplete adjuvant is drawn into a second glass syringe and, again, all air is evacuated. The two syringes are connected by a double Luer-lok adapter with a small central aperture. The two solutions are emulsified by injecting back and forth between the two syringes for 5 min. A milky white viscous emulsion results. When emulsification is complete, a single drop placed in water will not show any dispersion. RESULTS
Multiple trials with various dosages and concentrations of histamine were tried in dogs in an attempt to produce gastric secretion for 24 hr without side effects. An intramuscular dosage of 10 mg/day in a concentration of 20 mg/ml was found to give the best gastric secretory response without side effects. Heidenhain pouch acid output, though tapering off with time, did last a full 24 hr with an average daily acid output of 90 METHODS mequiv. Table 1 demonstrates the volume Histamine was used in the dihydrochloride and milliequivalents of acid output at various form which is very hygroscopic and, times. Since acid output decreases with time therefore, must be desiccated immediately after injection, maximal stimulation can best prior to use. One gram of the desiccated be achieved by giving the injection apdihydrochloride contains 604 mg of histamine proximately 6 hr after feeding. Thus the base. The desired quantity is dissolved in a animal’s endogenous gastric secretory output volume of saline appropriate to make a solu- in response to the meal is followed by the tion twice the actual desired concentration. long-lasting exogenous stimulus of the This solution is drawn into a glass syringe emulsified histamine solution. The ulcerogenic potential for emulsified histamine was tested in two groups of 10 ‘This work has been supported by National Institutes dogs each. In Group I, resection of the of Health Grant No. AM-04178. ‘Deceased. gastric antrum was performed with re357 Copyright o 1976 by Academic Press, Inc. All rights of reproduction in any form reserved.
358
JOURNAL
OF SURGICAL
RESEARCH:
TABLE 1 Histamine-Stimulated Gastric Secretion’ Time W 2 4 6 8 10 12 14 16 18 20 22 24
5mgat 50 mg/ml 16.2 7.4 6.2 5.6 1.1 0.5
IOmgat 20 mg/ml 9.5 13.6 12.3 11.3 8.8 9.5 7.8 6.5 4.3 3.0 1.4 1.2
15mgat 20 mg/ml 12.6 14.3 14.5 12.2 10.7 9.5 6.2 5.0 2.0 I .4 1.3 0.8
“All values expressed in milliequivalents per 2-hr collection period. The 5 and IO-mg doses had no associated ill effects. The 15-mg dose was always accompanied by nausea and/or diarrhea.
construction using the Billroth II method with closure of the duodenal stump and endto-side gastrojejunostomy. In Group II, the antrum was resected and the duodenal stump was closed, but reconstruction was performed with a Roux-en-Y gastrojejunostomy. The animals were permitted a recovery period of 3 weeks, at which time food intake, nutritional state, and body weight were stabilized. The emulsified histimine injections were then begun. In Group I (gastrojejunostomy), marginal ulcers appeared in 3 to 31 days in 4 of the 10 animals (40%). In the Roux-en-Y group, as one would expect from the classic experiment of Mann
VOL. 21, NO. 5, NOVEMBER
1976
and Williamson [3], the incidence of marginal jejunal ulcer was 100%. Surviving animals were all sacrificed at the end of 31 days. SUMMARY
AND CONCLUSIONS
A method has been described for the emulsification of crystalline histamine in incomplete Freund’s adjuvant. Intramuscular injection results in slow release with strong stimulation of gastric secretion over a period of 18 to 24 hr. The ulcerogenic potential for this method was established in two groups of dogs. In dogs with antrectomy and Billroth I1 gastrojejunostomy, 40% developed a marginal ulcer with a dose of 10 mg of histamine base per day. In a comparable group of antrectomized dogs with reconstruction by Roux-en-Y gastrojejunostomy, diversion of alkaline duodenal fluids from the stoma increased the ulcerogenic potential of chemical gastric secretory stimulation and 100% of the animals developed marginal jejunal ulcer. REFERENCES Hay, L. J., Varco, R. L., Code, C. F., and Wangensteen, 0. H. The experimental production of gastric and duodenal ulcers in laboratory animals by the intramuscular injection of histamine in beeswax. Surg. Gynecol. Obsfet. 75:170, 1942. Freund, J. The effect of paraffin oil and mycobacteria on antibody formation and sensitization: A review. Amer. J. Clin. Pathol. 21:645, 195 1. Mann, F. C., Williamson, C. S. The experimental production of peptic ulcer. Ann. Surg. 77:409-422, 1923.
