SOME ACUTE EFFECTS OF X-IRRADIATION (LDl00) o n

PLASMA AND ADRENAL TISSUE HISTAMINE IN RATS

APPROVED:

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Professor

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Minor Professor

MIA - ~K/ SK. Committee Member

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Director of the Departmeny jjf Biological Sciences

Deari of the Graduate School

Ferguson, James L. Some acute effects of X-irradiation

X L D i o o l O R Plasma and adrenal tissue histamine in rats.

Master of Science (Biology), May, 1972, 34 PP»? 6 figures,

3 tables, bibliography 43 titles.

The effects of a lethal dose (1380 r) of X-irradiation

on plasma and adrenal tissue histamine levels of rats were

studied. Histamine levels (determined fluorometrically),

adrenal weights, and blood counts were made at 1, 3, S, 9,

and 24 hours post-irradiation. X-irradiation was delivered

from a G. E. beryllium window X-ray unit at 120 KVP, 5 ma

with a | mi A1 filter at a target distance of 30 cm.

The plasma histamine response was triphasic (increase

at 1-3 hours, decrease at 5 and 9 hours and return to control

at 24 hours post-irradiation). The adrenal tissue histamine

response was found to be biphasic (decrease at 1 to 9 hours

and a return to control level at 24 hours post-irradiation).

A slight, but sustained, hypertrophy of the adrenal

glands was noted. A sustained severe eosinopenia occurred

while a slight leucocytosis was observed at the third hour

post-irradiation. No change in erythrocytes was noted.

The data suggests that histamine may be involved in

the action of the adrenal-pituitary axis following X-irradiation.

SOME ACUTE EFFECTS OF X-IRRADIATION (LD10Q) ON

PLASMA AND ADRENAL TISSUE HISTAMINE IN RATS

THESIS

Presented to the Graduate Council of the

North Texas State University in Partial

Fulfillment of the Requirements

For the Degree of

MASTER OF SCIENCE

By

James L. Ferguson, B. S,

Denton, Texas

May, 1972

TABLE OF CONTENTS

Page

LIST OF TABLES iv

LIST OF ILLUSTRATIONS v

INTRODUCTION 1

MATERIALS AND METHODS 6

RESULTS 12

DISCUSSION 23

SUMMARY 2 9

LITERATURE CITED 31

xix

LIST OF TABLES

Table Page

I. Effect of 1380 r Whole-body X-irradiation on Adrenal Weights 16

II. Effect of 1380 r Whole-body X-irradiation on Plasma Histamine Levels 19

III. Effect of 1380 r Whole-body X-irradiation on Adrenal Histamine Levels . . . . . . 21

xv

LIST OF ILLUSTRATIONS

Figure Page

1. Histamine Elution Columns 10

2. Effect of 1380 r Whole-body X-irradiation on Erythrocyte and Leucocyte Counts . . . 13

3. Effect of 1380 r Whole-body X-irradiation on the Eosinophil Count 14

4. Effect of 1380 r Whole-body X-irradiation on Mean Adrenal Weights, Per Pair . . . . 15

5. Effect of 13 80 r Whole-body X-irradiation on the Plasma Histamine Level 18

6. Effect of 1380 r Whole-body X-irradiation on the Histamine Content of Adrenal Glands 20

INTRODUCTION

Ellinger (12, 13) proposes that radiation sickness can

be classified as a typical case of the General Adaptation

Syndrome of Selye. This syndrome as described by Selye (37)

states that any severe change of an animal's environment may

initiate compensatory actions by the animal. This compen-

satory action is the unspecific response of the body to stress,

disregarding the causative agent. Such factors as cold,

electrical stimulation, bacterial toxins, ionizing radiation,

and other stressors have been employed experimentally to

demonstrate the adaptive defense mechanisms of an animal.

These defense mechanisms, according to the Selye concept, are

triggered by the stimulation of the hypothalamic centers, which

results in the stimulation of the anterior pituitary gland to

effect the secretion of tropic hormones.

Bacq and Fischer (9) reported an increased output of ACTH

in X~irradiated animals. Such increases in ACTH cause the

cortex of the adrenal gland to discharge an increased quantity

of adrenocorticosteroid hormones into the blood (the mineralo-

corticoids and the glucocorticoids). The mineralocorticoids

act mainly on the equilibrium of sodium and potassium in the

body, while the glucocorticoids act on the carbohydrate, pro-

tein, and lipid metabolism (8).

Schayer (36) implicated the adrenal cortex as a possible

site for histamine regulation by showing that glucocorticoids

have an inhibitory effect on the formation and binding of new

histamine in rat skin tissues. Moreover, in the case of rat

skin, he suggests that the action may be at any point in the

complex chain of events which follows the entry of free his-

tidine into the circulation of the skin and terminates with

the decarboxylation and binding of the histidine. This

decarboxylation and binding is thought to occur in mast cells,

Telford and West (40, 41) state that after injections of

corticoids into rats the histamine content of skin and small

intestine was reduced while that of the stomach tissue was

increased. The extent of change of histamine was found to

be roughly proportional to the glucocorticoid activity.

When administered to animals, histamine provokes respon-

ses which are typical of the so-called "Alarm Reaction" of

the stress concept proposed by Selye (37). This concept in-

cludes three pituitary-adrenal reactions: (a) alarm phase,

during which the adrenal gland undergoes hypertrophy and

increases secretion; (b) resistance phase, during which the

animal increases resistance to the stress; (c) exhaustion

phase, which terminates with the death of the animal if the

stress is too great. The development and duration of each

of the phases are dependent on the type and intensity of the

stressor that is applied. The alarm phase is characterized'

by the inhibition of growth, adrenal hypertrophy, thymus-

lymphatic atrophy, eosinopenia, and a depletion of the ascorbic

acid content of the adrenalglands, as reported by Fortier (17).

Fortier also indicated that the discharge of ascorbic acid by

the injection of histamine is indirect through stimulation of

the pituitary gland, because hypophysectomy abolished such an

effect. Gordon (19) suggests that the discharge of ascorbic

acid from the adrenals initiated by a small dose of histamine

was extremely rapid and was not prevented by a previous in-

jection of cortico-adrenal extracts.

Hameed and Haley (22) have reported a biphasic increase

of plasma and adrenal corticosterone in the rat at 2.5 and

48 hours post-X-irradiation with 650 r. Comparable data of

biphasic adrenal responses after whole-body X-irradiation

have been reported by a number of workers; however, the se-

cond response is more variable with respect to onset and

magnitude (16, 26). The synthesis and secretion of the adreno-

corticoids is thought to be dependent upon a neuro-endocrine

mechanism. This neuro-endocrine mechanism has been called by

Bacq and Alexander (8) the adrenal-pituitary axis. This

adrenal-pituitary axis regulates the production of ACTH, which

in turn controls the release of the adrenocorticoids. Neither

ACTH nor the adrenocorticoids have been shown to bring about

the liberation of histamine or affect its action after it is

released. However, Carryer and Code (10) indicate that both

the pituitary hormone (ACTH) and the glucocorticoids of the

adrenal glands may inhibit the biogenesis of histamine.

Although much has been written to indicate that there is

a close correlation between the conditions of shock and the

liberation of histamine from damaged tissue, Prosser et al.

(32) were the first to show an increase in histamine release

from tissues of animals exposed to mid-lethal to lethal doses

of X-irradiation in the first few hours after exposure. Haeger

et al. (21) have shown a leucocytosis in the rat to occur two

to twenty-four hours after X-irradiation. However, with this

leucocytosis a drastic reduction in the absolute eosinophil

count has been observed by Lott and Gaugl (26, 27) following

X-irradiation. Archer (5) has explained the eosinopenia that

is observed in rats under stress conditions on the basis of

changes in free histamine found in the tissues. Patt and his

associates (30) studied adrenal weight and histamine tolerance

in the rat and observed changes that seemed to be dependent

upon radiation dose and time after irradiation. However,

Wilson (43) noted that the histamine blood level in adrenal-

ectomized animals is lowered and resistance to injected

histamine is raised by adrenocortical extracts.

Although the role of histamine in the inflammatory process

is now relatively well known, its role in the radiation syn-

drome has not been fully elucidated. Since, as seen in the

foregoing review, metabolism of histamine appears to be closely

associated with the adrenal gland, it would seem feasible that

an attempt to correlate histamine changes in irradiated animals

with certain changes in the adrenal-pituitary axis ought to

be made. Such was the general aim of the present study.

The specific aims of the present study were to determine

the effect of lethal dosages of X-irradiation upon levels of

histamine in the blood plasma and adrenal tissues and see if

these histamine changes could be used to explain the initial

shock reaction (within 24 hours) associated with the radiation

syndrome. The parameters measured were erythrocyte count,

leucocyte count, absolute eosinophil count, adrenal weights,

blood plasma histamine, and adrenal tissue histamine.

MATERIALS AND METHODS

Male albino Sprague-Dawley rats of an average weight of

204 + 22.6 gm were used in this study. The stock animals were

supplied by the Ferguson Laboratory Animal Supply, Inc.,

Lewisville, Texas. Animals were kept in a community cage in

a temperature-controlled animal house which was continually

lighted. The lights were left on constantly to try to negate

any diurnal histamine level changes.

The rats were fed Wayne Dog Chow and tap water ad_ libitum,

Following X-irradiation, the rats were transferred to the

laboratory and maintained there until the designated time of

sacrifice. In all of the test experiments, irradiation and

testing were performed in groups of three to five animals.

Filtered X~irradiation mm aluminum filter) was de-

livered from a General Electric beryllium window unit at 120

KVP and 5 ma. All test rats in this study received a lethal

dose of 1380 r (138 r/min). The target distance was 30 cm,

with an exposure time of 10 minutes. To insure uniform ex-

posure, all X-irradiated rats were placed in a plexiglass

tube-motor apparatus and rotated at the rate of one rpm. The

sham-irradiated rats were treated identically to the test

rats.

This study was divided into three series: (a) pre-tests,

(b) sham-irradiated series, and (c) X-irradiated series. A

minimum of six rats was used for each sampling time of the

sham-irradiated series. Eighteen rats were used for the

resting controls. The sham-irradiated and test animals were

sacrificed at predetermined intervals of 1, 3, 5, 9, and 24

hours after X-irradiation or sham-irradiation. To check the

effects of rotation, another group of rats, the pre-test

series, was taken directly from the animal quarters and sac-

rificed after being rotated, but not sham-irradiated or

X-irradiated. Six physiological parameters were monitored

during this study: (a) red blood cell count, (b) white blood

cell count, (c) the absolute eosinophil count, (d) the weights

of decapsulated adrenal glands, (e) the plasma histamine, and

(f) the adrenal tissue histamine. The red and white cell

counts were made from tail samples obtained after the rats had

been anesthetized with intraperitoneal injections of Nembutal

(33 mg/gm body weight). To aid in the blood sampling, each

rat's tail was immersed in warm water. Following blood sam-

pling from the tail, the animals were immediatedly guillotined

and more blood samples taken for the plasma histamine determi-

nations . The adrenal glands were then removed and decapsulated,

weighed (mg/pair/100 gm weight^ and stored for tissue histamine

analysis.

The red and white blood cell counts were made using the

direct-chamber counting method. Absolute eosinophil counts

were made following the direct-chamber counting method of

Pilot (31).

-8

Anton and Sayre1 s (1) modification of the Shore ejb al.

(38) method was employed to determine the adrenal tissue

histamine in this study. The procedure is outlined as follows:

The paired adrenals were homogenized in 2.0 ml of 0.4 N HCIO^

in a glass homogenizer submerged in ice, and the homogenate was

transferred to a 12 ml pyrex centrifuge tube. One-half ml of

H2O was added with thorough mixing and centrifuged at 10,000

rpm in a refrigerated centrifuge for 15 minutes. Two ml of

the supernatant were transferred to another centrifuge tube

containing 2.0 ml of H2O and mixed. The dilute supernatant

fraction was then poured into a 50 ml pyrex centrifuge tube

containing 3.5 gm of K2PHO4 and mixed. Twenty ml of isoamyl

alcohol were added, the mixture was mechanically shaken, and

centrifuged at 3,000 rpm for 5 minutes to separate the phases.

Twenty ml of the organic layer were pipetted into another 50

ml pyrex centrifuge tube containing 4.0 ml of 0.01 N HC1 plus

15 ml of heptane. The mixture was mechanically shaken for 6

minutes and then centrifuged for 5 minutes at 3,000 rpm after

the organic phase was aspirated and discarded. Three and

one-half ml of the acid extract were pipetted into another

centrifuge tube containing 0.5 ml of 10 N NaOH and mixed. To

this mixture 2.5 gm of NaCl and 20 ml of CHCl^ were added and

mechanically shaken for 3 minutes, followed by centrifugation

at 3,000 rpm for 3 minutes to separate the phases. The washed

extract (top layer, 4.0 ml) was then pipetted into another

centrifuge tube to which 1.5 gm of NaCl were added and thoroughly

9

mixed. To this mixture were added 20 ml of isoamyl alcohol,

and the resultant mixture was mechanically shaken for 6 min-

utes and centrifuged at 3>000 rpm for 5 minutes. Twenty ml

of the organic phase were pipetted into another pyrex centri-

fuge tube containing 2.0 ml of 0.1 N HC1 plus 15 ml of heptane

and this mixture was shaken for 6 minutes, centrifuged at

3,000 rpm for 5 minutes, and the organic layer aspirated and

discarded. The aqueous layer was saved for analysis.

Fluorophor formation was accomplished by adding the fol-

lowing reagents: 0.25 ml of sample, 0.25 ml of 0.1 N HC1,

1.10 ml of 1.0 N NaOH, 0.225 ml of OPT reagent, wait 4 minutes,

and add 0.05 ml of 2.0 M citiric acid. The fluorophor was then

measured in a Turner fluorometer, model 110. The fluorometer

was equipped with a No. 7-60 primary filter and a No. 2A sec-

ondary filter. The data are expressed in mg of histamine per

gm of adrenal tissue.

Noah and Brand's (29) procedure was used to determine

plasma histamine levels of the animals in this study. This

technique, also a modification of the Shore et ajl. (38) method,

employs glass elution columns which have two constrictions

with the following dimensions: total length 160 mm, 22 mm

opening, 3*5 mm internal diameter at the first constriction,

which is 50 mm below the opening, 1 mm internal diameter at

the second constriction, which is 120 mm below the opening

(See Fig. 1). The columns were packed with 50 mg of cellulose

acid succinate, (CAS).

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The CAS was prepared by mixing 1.25 gm anhydrous sodium

acetate, 10 gm of succinic anhydride, 75 ml of glacial acetic

acid, and 2.5 gm of cellulose all in a heavy-walled pyrex

bottle which was stoppered with a drying tube. The bottle

was then placed in an oven at 100°C. for 48 hours and removed

occasionally for gentle shaking.

The CAS was filtered on filter paper with gentle suction.

Then the CAS was washed with 500 ml of water, 500 ml of 1%

HC1, and again with 1,000 ml of water. The washed CAS was

resuspended in water, filtered, and washed with water. Then

the CAS was dried with 95%> ethanol, absolute ethanol, and

then in a vacuum oven at 50°C. for 3 hours. The CAS was then

ready for packing the columns. After the columns were packed,

the packing was used for ten extractions before being replaced

with new CAS. The columns were cleaned by washing them with

95% ethanol and absolute ethanol between extractions. After

fluorophor formation from the extracts from the columns the

plasma histamine concentration was stated in terms of aig/L.

RESULTS

In the graphs each circle represents the mean value of

at least six animals.. The letter "C" represents unrotated

animals that were brought directly from the animal quarters

and sacrificed. The pre-test animals "were sham-irradiated

animals that were brought from the animal quarters and sac-

rificed after they had been rotated ten minutes in the plexi-

glass chamber.

Figure 2 depicts the effects 1380 r X-irradiation on the

total erythrocyte (A) and leucocyte (B) count in rats. It is

clear that such dosage did not alter the red blood counts

during the 24 hours post-irradiation. On the other hand a

significant increase in leucocyte count was observed at the

3rd hour post-irradiation. Following a return to control levels

at the 5th and 9th hours post-irradiation a significant de-

crease in leucocytes was noted at the 24th hour post-irradiation.

As shown in Figure 3, the eosinophil response to a lethal

dose (1380 r) of whole-body X-irradiation was striking. It

was noted that there was an immediate and sustained eosinopenia

in the irradiated animals. The sham-irradiated animals did

not vary markedly from the resting controls or pre-test animals

in their absolute eosinophil counts.

Figure 4 and Table I show the effects of a lethal dose

(1380 r) of X-irradiation on the paired wet adrenal weights.

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From the data shown here it is evident that there was a sus-

tained hypertrophy of the adrenal glands after whole-body

exposure to X-irradiation. The data, summarized in Table I,

were found to be statistically significant regarding the in-

crease in adrenal wet weight at all sampling times post-

irradiation. It was also evident that the observed hypertrophy

appeared to become larger with time. The sham-irradiated

groups showed no significant change from the pre-test series.

The effects of a lethal dose (1380 r) of whole-body

X-irradiation on plasma histamine may be seen in Figure 5 and

are summarized in Table II. As seen in Figure 5, the plasma

histamine concentration showed a significant rise from the

initial concentration at the third hour post-irradiation.

However, the histamine level dropped to sub-control levels at

the fifth and ninth hour post-irradiation. Recovery in the

histamine level occurred at the twenty-fourth hour post-

irradiation. The sham-irradiated animals were at no time

significantly different in plasma histamine levels from the

pre-test series.

As shown in Table II, the differences in the means of the

histamine levels observed in all of the X-irradiated animals

were statistically significant from the means of the pre-test

series.

The effects of 1380 r X-irradiation on adrenal tissue

histamine are depicted in Figure 6 and summarized in Table

III. In Figure 6 it can be seen that there was a decrement

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22

in histamine level in the adrenal tissues at 1, 3, 5, and 9

hours post-irradiation. The decrease, however, as indicated

in Table III, was significant from the means of the pre-test

series only at 1, .3, and 5 hours post-irradiation. At 9 and

24 hours post-irradiation, the differences observed between

the test series and the sham-irradiated controls were not

significantly different.

If one compares the data in Figures 5 and 6, it may be

noted that there was little correlation between the changes

in tissue histamine and circulating histamine. X-irradiation

brought about an immediate and relatively sustained decrease

in adrenal histamine. On the other hand, the same dosage of

X-irradiation brought about a biphasic response in re circu-

lating histamine levels; ici est, and initial enhancement fol-

lowed by a decrease in histamine. Interestingly, recovery

to control levels were noted in both parameters at the same

time: id est. 24 hours post-irradiation.

DISCUSSION

The experimental procedures used in this study for

determining plasma (Noah and Brand, 29) and adrenal tissue

(Anton and Sayre, 1) histamine levels were both modifications

of the method of Shore et al. (38). Multiple assays were

carried out in an effort to give statistical validity to the

use of these newer extraction and analysis procedures for

histamine. Preliminary trials using these modified methods

resulted in recoveries of eighty-nine percent for Noah and

Brand's method for plasma histamine and eighty-six percent

for Anton and Sayre's method for tissue histamine. It was

concluded that the newer methods yielded more valid histamine

determinations than Shore's procedure.

Regarding the erythrocyte data, Bacq and Alexander (8)

stated that following LD-^QQ radiation anemia will develop if

the animal lives long enough. However, the data here indicate

that this anemia did not develop within the first twenty-four

hours post-irradiation. This finding was not unexpected since

Supplee et al. (39) have shown that, due to the 80-120 day

turnover time of erythrocytes in the rat, anemia would not

likely appear until late in the radiation syndrome. Although

the erythrocyte count may not be directly related to hista-

mine concentration of the blood or plasma, Fulton et al, (18)

have indicated that after whole-body irradiation there is an

23

24„

increased extracellular fluid and plasma volume without a

concomitant decrease in the total red blood cell count.

Archer, in his monograph (6) on the role of eosinophils

in the body, has presented evidence that almost all of the

blood histamine is contained in the white blood cells and

particularly in the eosinophils. Indeed, he stated that the

eosinophil was the main carrier of blood histamine. Code

(11), however, believes that the basophil leucocyte, like the

tissue basophil mast cell, is an important storage place for

histamine. Samson and Archer (35) presented data to show that

the basophil contains significant amounts of histamine while

other white cells do not. However, they admit that the hista-

mine content of human basophils was much lower than the

histamine content of rat mast cells.

The role of certain leucocytes as histamine liberators has

been reported by Lutz et al. (28). They noted that two basic

peptides in lysosomes of normal bovine polymorphonuclear leuco-

cytes were potent histamine liberators. These histamine-liberating

peptides were found to contain high amounts of arginine and

demonstrated more effective histamine liberating properties when

there was an alternative sequence of arginine and glycine. They

further suggested that these arginyl peptides increased vascular

permeability through mast cell disruption.

Riley and West (33) have presented evidence that tissue

mast cells are the likely site for the intracellular location

of much of the histamine found in rats. According to Archer

25

(2,3) and Archer and Jackas (4)* the disruption of eosino-

phils causes the release of eosinophil peroxidase. This peroxi-

dase initiates the lysis of the basophil granule. Both the dis-

ruption of eosinophils and lysis of basophils could result in

an increased free histamine. Lott (25) presented data showing

that there was a sharp and sustained eosinopenia after exposure

to sublethal and lethal dosages of X-irradiation. If this drop

in eosinophils in fact was due to lysis of these cells, then

they would release the peroxidase which in turn would result in

lysis of mast cells, initiating a release of histamine into the

plasma or tissues. Such action might account for the initial in-

crease in the plasma histamine noted in this study. However, it

does not explain the secondary decrease in plasma histamine and

then the return to control levels that followed. One might con-

jecture that there would have to be another mechanism besides

the eosinophil lysis (decreased binding?) to account for the

return of histamine to control levels. The hypothesis of

Archer (5) that eosinophils inactivate histamine cannot be dis-

regarded on the basis of the present data; however, his hypothesis

would be strengthened if the eosinophil response had followed the

plasma histamine response after X-irradiation.

When administered to animals, histamine elicits responses

which are typical of the so-called "Alarm Reaction" of Selye

(37). After administration of histamine, Fortier (17) has

shown that there is an inhibition of growth, adrenal hyper-

trophy, thymus-lymphatic atrophy, eosinopenia, and a depletion

26

of the ascorbic acid content of the adrenal glands. Indeed,

adrenal hypertrophy was one of the earliest used indices of

adrenal activity during stressful conditions. Patt et aJL. (30)

have shown unilateral hypertrophy to follow six hours after

exposure to X-irradiation. The slight adrenal hypertrophy

that was observed in the present study might be explained on

the basis of Selye's (37) General Adaptation Syndrome. In

the alarm phase an animal that has been exposed to a stress

agent demonstrates eosinopenia, a hypertrophy of the adrenal

gland, and an increased secretion, supposedly due to the influ-

ence of ACTH, Indeed eosinopenia has been used as a bioassay

tool for ACTH.

Arora and Lahiri (7) have presented data to show that

the metabolism of histamine is intimately associated with the

adrenal glands. Rose and Brown (34) have demonstrated that

adrenalectomized animals become very sensitive to the toxic

effects of histamine. After adrenalectomy the histamine level

rises in the blood as well as in several other tissues; for

example, the gut lining. Moreover, Houssay (23) has demon-

strated that hypophysectomy also sensitized animals to the

toxic action of histamine, although in a lesser degree than

adrenalectomy. Guillemin (20) and Fleming and Geierhass (15)

have shown that an increased sensitivity to histamine is not

only an indication of an increase in histamine, but may also

be due to a lack of corticoids and a deficiency in body cate-

cholamines. In this respect, whole-body irradiation has been

2'7

shown by a number of workers (14, 16, 22, 26) to stimulate a

biphasic plasma corticoid response in rats. The first ele-

vation in the glucocorticoids observed in this study at 5

hours post-irradiation might be due to the depressing action

of the glucocorticoids on histamine levels. Thus, when the

circulating corticosterone is at a high level, the histamine

level decreased in the plasma.

The initial increase in plasma histamine that was noted

at 1 and 3 hours post-irradiation fits very well with Weber

and Steggerda's (42) data on blood pressure drops in rats at

the second and third hours post-irradiation. The very effec-

tive vasodilator, histamine, has been shown to cause a pro-

nounced drop in blood pressure when injected into the blood

stream. The blood pressure in X-irradiated rats has also

been shown to return to normal after the first reaction. This

would seem to follow very closely the plasma histamine re-

sponse to whole-body X-irradiation noted in this investigation.

This study has shown that the irradiated animals demon-

strated a significant and sustained decrease in adrenal

histamine at 1, 3, and 5 hours post-irradiation. This decrease

occurred during the same period at which the plasma histamine

was at its highest level after irradiation. One must remember,

however, that the relative concentration of the tissue hista-

mine is drastically higher than that found in the plasma.

This is probably due to the greater number of mast cells in

the tissues as compared with the blood, or it may be due to

28

changes in the release and/or binding of histamine in the

blood and tissues. The diminution of histamine of the

adrenal glands might be explained by the observations of

Leinweber and Braun (24). These workers found that there

is a suppression of histidine decarboxylase activity of rat

tissues under moderate to severe stress. With this sup-

pression there would be a decreased production (release?)

of histamine from histidine in the tissues. On the other

hand, the initial rise in plasma histamine may be due to

the fact that X-irradiation brings about a faster release

and/or production of histamine than corticoid production:

therefore, the depressant action of the corticoids could

not be manifested until they were in sufficient quantities

by the 3-5 hours post-irradiation.

The early symptoms in the radiation syndrome are similar

to that of the histamine response in some animals. One might,

therefore, conjecture that when the histamine levels were

decreased in both the blood and adrenal tissues post-radiation

and during an eosinopenic period, one would expect to see a

pronounced histamine response by the animal in toto. at least

clinically.

The question then arises as to whether one might use

plasma histamine analysis as a radio-biological indicator

tool and/or a prognostic tool in dealing with animals, in-

cluding humans, exposed to ionizing radiation.

SUMMARY

This study concerned the effects of a lethal dose

(1380 r) of X-irradiation on the histamine response in rats.

Parameters measured were (1) plasma and adrenal tissue

histamine determined fluorometrically; (2) adrenal weights;

(3) circulating eosinophils using Pilot's method; and (4)

total red and white blood cell counts. Blood samples were

taken prior to and at various intervals post-irradiation.

It was found that

(1) the plasma histamine response was triphasic, id est

an increase at one and three hours post-radiation,

a decrease at five and nine hours post-radiation,

and a return to control levels by the twenty-fourth

hour post-radiation;

(2) the adrenal tissue histamine response, on the other

hand, was biphasic, id est a decrease at the one,

three, five, and nine hours post-radiation and a

return to control levels at the twenty-fourth hour;

(3) a slight, but sustained, hypertrophy of the adrenal

glands occurred;

(4) an immediate, pronounced, and sustained eosinopenia

developed;

(5) no change in erythrocyte count was evident; and

29

30

(6) a slight leucocytosis was observed at the third

hour post-radiation.

The rise in plasma histamine was explained on the basis

of a possible delayed corticoid production. The observed de-

crease in adrenal histamine was explained on the basis of a

possible depression of histidine carboxylase activity in var-

ious tissues and cells including eosinophils and mast cells.

The data in this study indicate (1) a definite histamine

response to X-irradiation that may be used as a radio-biological

indicator tool; (2) that the initial symptoms in the radiation

syndrome may be histaminic in nature; and (3) that histamine

may play an important role in the action of the adrenal-

pituitary axis following X-irradiation.

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