SOME ACUTE EFFECTS oOF X-IRRADIATION (LD l00/67531/metadc163992/m2/1/high_res_d/n_04448.pdfduring...
Transcript of SOME ACUTE EFFECTS oOF X-IRRADIATION (LD l00/67531/metadc163992/m2/1/high_res_d/n_04448.pdfduring...
SOME ACUTE EFFECTS OF X-IRRADIATION (LDl00) o n
PLASMA AND ADRENAL TISSUE HISTAMINE IN RATS
APPROVED:
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Professor
f;
Minor Professor
MIA - ~K/ SK. Committee Member
^ J ' t\ < X r-g.
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).
10
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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.
12
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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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