Upsala J Med Sci 84: 247-254, 1979 

The Effects of Endogenous Hypergastrinemia and 
Hypogastrinemia on the Exocrine and 

Endocrine Rat Pancreas 
H. Johansson'.2, L. Grirnelius3, Ph.U. Heitz', G. Lundqvist4, 

P. Peterson5, G. Portela-Gomes3 and E. Wilande? 
From the Depurtments of Surgery', Clinicul Research 112, Pathology3, and Clinicul Chemistry4, 

University Hospital, Uppsala, the Department of Histology5, Uppsala University, Uppsalu, 
Sweden and the Department of Pathology6, University of Basel, Switzerland 

ABSTRACT 

The effects of endogenous hypergastrinemia and hypogastrinemia on the exo- 

crine and endocrine pancreas were studied in the rat. Hypergastrinemia was in- 

duced by antral exclusion, and hypogastrinemia by antral resection. The studies 

were made 14 weeks after surgery. 

The total weight of the pancreas was increased both in hypergastrinemic and 

hypogastrinemic animals, due to hypertrophy of the exocrine cells. In contrast, 

the volume and total weight of the pancreatic islets were decreased. 

There was no numerical difference in the A-, D-, PP-cells between the hyper- 

and hypogastrinemic animals, respectively, and the controls. The number of in- 

sulin-producing (B-) cells was certainly reduced after the induction of hypo- 

gastrinemia. There was, however, signs of increased B-cell activity, which 

might contribute to an underestimation of the number of B-cells with the tech- 

nique used. 

These findings do not support the hypothesis that antral gastrin has trophic 

influence on either exocrine or endocrine pancreas. 

INTRODUCTION 

Gastrin is known to influence the gastrointestinal tract in many ways. Its 

stimulatory action on the secretion of hydrochloric acid and pepsinogen and its 

trophic effect on the parietal cell area of the stomach are well defined ( 3 , 1 9 ,  

2 0 ) .  Pentagastrin, administered in pharmacological doses, has also been re- 

ported to exert a trohpic action on the exocrine pancreas in rats ( 1 , 9 ) .  

Hitherto, however, no such action has been found following various surgical 

procedures leading to an increased endogenous serum gastrin concentration ( 1 3 ) .  

In the experiments by the latter authors no increase in the total dry weight of 

the pancreas was observed. A link between gastrin and the endocrine pancreas 

has been discussed by Larsson ( l o ) ,  who found islet cell hyperplasia in some 

patients with hypergastrinemia due to tumor production and in patients with 

atrophic gastritis. Furthermore, indirect evidence of a gastro-insular axis 

247 



mediated by gastrin has been produced, as an altered glucose tolerance has been 

found in patients with hypergastrinemia compared to normogastrinemic patients 

(16,17). 

The present experiments were designed in order to further examine the effect 

of endogenous hypergastrinemia not only on the exocrine but also on the endo- 

crine pancreas. 

MATERIAL AND METHODS 

Adult male Sprague-Dawley rats (body weight 250-300 9) were used. They were 

fed on laboratory food. The animals were randomly assigned to the following 

g roups : 

(1) 5 rats with exclusion of the antral part of the stomach (AE) 

( 2 )  4 rats with resection of the antral part of the stomach (AR) 

( 3 )  4 control rats - untreated ( C ) .  
Antral exclusion and antral resection were performed by techniques described 

previously ( 5 ) .  

Serum gastrin concentrations were determined by solid-phase radio-immuno- 

assay ( 1 1 )  12 weeks after operation, after a starvation period of 48 h. 

The animals were killed 14 weeks after the stomach operation. The pancreas 

was extirpated, dissected free from fat tissue and weighed. The specimens were 

then fixed in Bouin‘s fluid for 18 h, dehydrated, cleared in xylene and em- 

bedded in paraffin. Deparaffinized 5 um thick sections were stained with hema- 

toxylin-eosin and by the van Gieson method. 

To calculate the percentage volume of the pancreatic islets, the point sam- 

pling technique described by Chalkley ( 2 )  was employed. At least 3000 inter- 

sections (hits) were analysed on acinar or islet parenchyma of each pancreas. 

The weight of the endocrine parenchyma was calculated by multiplying its per- 

centage volume by the weight of the entire pancreas. The densities of the exo- 

crine and endocrine tissue were assumed to be equal. 

The density of the exocrine cells per unit volume exocrine tissue was deter- 

mined in a microscope equipped with a square grid placed in the focal plane of 

the eyepieces at a magnification of x 400. In each case three randomly selected 

areas of exocrine tissue were examined and the nuclei or nuclear fragments 

within the square grid were counted. The nuclear sizes of 50 randomly selected 

exocrine cells were also determined with the aid of a mm scale placed in one 

of the eyepieces. As the observed nuclear frequency is influenced by the nu- 

clear size, the formula of Flodbrus was used to correct these values (7). The 

relative number of exocrine cells was calculated by multiplying the corrected 

number of exocrine cells per unit volume by the exocrine pancreatic weight. 

Gomori’s aldehyde fuchsin technique (12) with ponceau fuchsin as a counter- 

stain (8) was used to visualize B-(insu1in)cells. D-(somatostatin)cells were 



stained with the Davenport alcoholic silver nitrate technique as modified by 

Hellerstrom & Hellman (6). A-(g1ucagon)cells and PP-(pancreatic polypeptide) 

cells were identified with the immunoperoxidase (PAP) technique described by 

Sternberger ( 1 8 ) .  The sections were counterstained with Hernechtrot. Controls 

a s  described by Goldman (4) were used. 

Differential counts were made in order to estimate the frequency of differ- 

ent types of endocrine pancreatic cells (except PP-cells). At least 2000 endo- 

crine cells were counted in each animal and with each of the various staining 

procedures. Only cells with visible nuclei or nuclear fragments were included 

in the counts. 

The nuclear size of B- and non-B-cells was estimated in aldehyde-fuchsin- 

stained sections (counterstained with ponceau fuchsin) as described above, at a 

magnification of 1 2 5 0  x. 

Statistics: Student’s t-test was used. 

RESULTS 

The mean body weight at sacrifice was slightly higher in the control group 

than in the two experimental groups (Table 1 ) .  

The serum gastrin concentration was considerably increased in group AE and 

markedly decreased in group AR as compared with that in the controls (Table 2). 

The mean pancreatic weight was much higher in both experimental groups than 

in the controls (Table 3 ) .  

The observed number of exocrine cell nuclei per unit volume was highest in 

the control group and lowest in group AR. After correction concerning the RU- 

clear size the differences were more pronounced (Table 4). These findings indi- 

cate a hypertrophy of the exocrine pancreas both after antral exclusion and 

after antral resection. The relative values of the total number of the exocrine 

cells were also highest in the control group and lowest in group AR (Table 4 ) ,  

indicating a numerical decrease of the exocrine cells after antral resection. 

The volume of the pancreatic islet tissue in relation to the total pancre- 

atic volume was significantly lower in the experimental groups (Table 5). The 

calculated weight of the islets was significantly decreased in both experimen- 

tal groups in comparison with the control group (Table 5 ) .  The islet weights of 

the two experimental groups did not differ from each other. The amount of non- 

parenchymal tissue (blood vessels, connective tissue, fat cells) was approxi- 

mately equal in the three groups as judged by visual observation. 

In group AE and the control group the B-cells showed a strong and even 

staining reaction, while this reaction was highly variable in group AR. No dif- 

ference in the staining of A - ,  D- or PP-cells was found between the different 

groups. The frequency of B-cells in group AR was significantly lower than in 

the controls (Table 6), whereas the frequency of the other endocrine cell types 

249 



Table 1. Body weight at sacrifice 

Group Body weight (4) 
M C S.D. 

AE 4 1 7  f 3 4  
AR 4 1 1  2 2 9  
Controls 4 6 3  f 3 5  

Table 2. Serum gastrin 

Group Serum gastrin (mmol/l) 

AE 2 8 7  ? 2 6  
AR 1 3 8  f 3 5  
Controls 1 7 0  f 1 9  

Table 3. Weight of the pancreas 

Group Weight of pancreas (9) 
M f SE 

AE 2 . 2 9 7  C 0 . 1 0 6  
AR 2.319 2 0 . 1 0 7  
Controls 1 . 9 8 2  C 0 . 1 1 5  

(A- and D-cells) did not differ significantly between the three groups. PP- 

cells were only seen in some islets, mainly located in the periphery. They 

varied in number, but were most often sparse. No differences in the distribu- 

tion were observed between the different groups. 

The diameters of the B- and non-B-cell nuclei (Table 7 )  did not differ sig- 

nificantly between the three groups. However, in group AR the nuclear diameter 

of the weakly stained B-cells was significantly greater ( 6 . 0 5  C 0 . 1 3 )  than that 

of the heavily stained B-cells ( 5 . 5  f 0 . 1 0 ) .  

DISCUSSION 

In these experimental models endogenous hypergastrinemia was induced by 

antral exclusion and hypogastrinemia by antral resection. The total pancreatic 

weight was higher in both experimental groups than in the control animals. This 

enlargement of the pancreas was due to hypertrophy and not to hyperplasia of 

the exocrine cells. The cellular hypertrophy was especially prominent in group 

AR. The hypertrophied cells were also characterized by nuclear enlargement, 

250 



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probably reflecting increased cellular activity. On the other hand the total 

volume of pancreatic islet tissue was decreased both after antral exclusion and 

after antral resection. These findings do not indicate a trophic effect of 

antral gastrin on the exocrine o r  endocrine pancreas, since similar changes 

were observed in the two experimental groups. 

When adding the percentages of the different cell types in the same group, 

the content exceeded 100 % for group AE and the controls. This was due to the 

fact that with the staining procedures used A-, B-, and D-cells are visualized 

in different ways. The reason for this was that the conditions for differential 

counting varied with staining technique, as not only the cell type in question 

but also other endocrine cells could appear differently depending on the back- 

ground counterstain. Nevertheless, a comparison of the frequency of a particu- 

lar type of cell between the individual groups is considered justifiable. 

The only difference noted between the two experimental groups was a lower 

frequency of B-cells in group AR, but on the other hand, the 3-cells in this 

group showed an increased rate of synthesis (nuclear enlargement) and release 

(decreased staining reaction to insulin). This lower frequency of B-cells might, 

however, be misleading, since some of the more or less degranulated cells might 

have been overlooked in the differential counting. The reason for this appar- 

ently altered function of B-cells after antral resection is not known. 

It must be emphasized than in these experiments the duodenum was excluded 

from continuity with the gastrointestinal tract after both surgical procedures. 

It is therefore possible that some duodenal factor or factors might have been 

involved in the pathogenesis of the observed changes in both the endocrine and 

the exocrine pancreas. Such factors might well include hormones, e.g. chole- 

cystokinin, secretin, somatostatin and duodenal gastrin. It has been suggested 

that exogenous cholecystokinin may exert a trophic effect on the exocrine pan- 

creas (14,15), while secretin and somatostatin are believed to counteract the 

trophic action of gastrin (13,21). In our study the secretion of these hormones 

was not analysed. However, it may be assumed that the secretion of cholecysto- 

kinin and secretin was decreased, as stimulation by intraluminal contents of 

the duodenum is excluded. In order to test these speculations further, studies 

on the possible trophic action of duodenal hormones in the rat are planned. The 

findings of the present study with the experimental model used do not support 

the view that antral gastrin has a trophic effect on the rat pancreas. 

ACKNOWLEDGEMENTS 

Pure HPP and anti-HPP serum were a generous gift from Dr R.E. Chance, Lilly 

The expert technical assistance of Miss M. Kasper is gratefully acknowledged. 
The study was supported by grants from the Swedish Medical Research Council 

Research Laboratories, Indianapolis. 

(Nos 1 0 2 ,  4534 and 2 2 9 7 ) .  

253 



REFERENCES 

1. Barrowman, J.A. & Mayston, P.D.: The trophic effect of pentagastrin on the 
pancreas. In: Gastrointestinal Hormones (ed. J.C. Thomson), pp. 231-247, 
Univ of Texas Press, Austin and London, 1975. 

2. Chalkley, H.W.: Method for the quantitative morphological analysis of 
tissues. J Nat Care Inst 4:347-353, 1943. 

3. Enochs, M.R. & Johnson, L.R.: Hormonal regulation of the growth of gastro- 
intestinal tract; biochemical and physiological aspects. In: Progress in 
Gastroenterology (ed. G.B.J. Glass), Vol. 111, pp- 3-28, Grune and Stratton, 
New York, 1977. 

York, 1968. 

mental study in the rat with special reference to gastrin and some hyper- 
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Received August 10, 1979 

Address for reprints: 

H. Johansson, M.D. 
Department of Surgery 
University Hospital 
5-750 14 Uppsala, Sweden 

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