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Effect of Histamine and of the Anaphylactic Reaction on
Isolated Guinea Pig Atria
By
MARIO PENNA, M.D.,
ALEJANDRO ILLAXES, M.D.,
SERGIO MUJICA,
MARIO UBILLA, M. D., AND
M.D.
Addition of specific antigen to the isolated atria of sensitized guinea pigs and of histamine to those of nonsensitized animals resulted in an increase of both frequency and
amplitude of contraction and in the appearance of rhythmic activity in the isolated left
atrium. On a quantitative basis the anaphylactic response looks like the effects of histamine
in concentrations between 10~3 and 10~2 mM/L. Xo qualitative differences between the
effects of histamine and those of the anaphylactic reaction were observed. Since isolated
atria thus are seen to participate in tissue sensitization, the heart should be considered
as a potential shock organ in the anaphylactic reaction.
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XPERIMENTS 1 - 3 performed during the
second decade of this century showed
that during induced anaphylaxis there were
disorders of cardiac frequency and rhythm
which were not prevented by vagotomy.
Electrocardiographic studies disclosed increases in heart rate, alterations in atrioventricular conduction, and changes in the
origin and propagation of the excitation
wave. The conditions prevailing in these experiments did not permit a decision on
whether these observed changes were due to
a direct effect of the anaphylactic reaction
upon the myocardium or were attributed to
myocardial ischemia and other alterations
in circulatory dynamics.
Wilcox and Andrus in 1939 4 - 5 studied
this question on the isolated heart. During
the anaphylactic reaction induced by the
addition of specific antigen they observed
sinus tachycardia, prolongation of P-R interval, A-V dissociation, nodal and ventricular
tachycardias, and other disorders of rhythm.
Since a decrease of coronary flow was also
present, the authors suggested that these
changes in rhythm might be the consequence
of an impairment of myocardial oxygenation.
Kellner, Penna and Schweid 0 reported that
the addition of minute quantities (0.02 to
2 /xg.) of the sensitizing antigen to the fluid
perfusing isolated guinea pig hearts, previ-
E
ously sensitized to crystalline streptococcal
proteinase, bovine y-globulin, or crystalline
egg albumin, resulted in a considerable increase in frequency, usually accompanied by
a marked increase in amplitude of contraction and certain disorders of rhythm (premature contractions, A-V dissociation, even
complete heart block). This reaction was
highly specific for the agent used to sensitize
the animal. Coronary flow was simultaneously measured and no consistent changes
were observed. Experiments on isolated atria
of sensitized guinea pigs, reported in the
same paper, showed reactions similar to
those observed in the whole isolated heart,
when such preparations were exposed to the
sensitizing agent in concentrations as low as
0.001 jug./ml.
Wilcox and Andrus5 studied the effects of
histamine and of the anaphylactic reaction
on the isolated guinea pig heart and observed
that both involved an increase in heart rate
arrhythmias, and a decrease of coronary
flow. They also reported that atropine antagonized the histamine effect on coronary
flow, but not the tachycardia or the arrhythmias, and that this drug did not alter the
anaphylactic reaction in the isolated guinea
pig heart.
In order to elucidate this phenomenon
further by avoiding changes in coronary flow
it seemed desirable to study tti£ effect of
histamine and of the anaphylactic reaction
in isolated atria while registering simultan-
From the Instituto de Farmaeologia, Escuela de
iledicina, Universidad de Chile, Santiago, Chile.
Received for publication November 28, 1958.
521
Circulation Research, Volume VII, July 1950
PENNA, ILLANES, UBILLA, MUJICA
522
TABLE 1.—Effect
of Histamine and of Anaphylactic
Effect of liistaminc
1 X 10"' 24
1 X 10"3 23
1 X 10- 25
1 X lO"1 20
+ 2.3± 1..5
+ 14.2± 1.7
+41.5± (i .0
+43.0±12..0
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Effect of antigen
Sensitized
guinea
12
+19.4± 4
pigs
Nonscnsitized
guinea
8
+ 0.5± 0.C}
Pigs
1
C
o
Appearance
of rhythmic
activity in
isolated left ntriuni*
amplitude
1
(per cent of initial)
Changes of
frequency
(per cent of initial)
Number of
experiments
Histaminc
concentration
(mM/L.)
Reaction on Isolated Guinea Pig Atria
+13.6± 2.05
+ 28,.3± 5.5
+ 68..1±15.9
+66. 6± 7.5
0/11
3/1.1
3 0/11
10/11
+40 ±17.1
5/8
— 8 ± 3.2
—
•Positive cases of total isolated left atrium.
eously the electrical and mechanical manifestations of the contractions. Since Highland Tiemann 7 reported that histamine induces
automaticity in the isolated left rabbit atria,
it was also decided to see if the auaphylactic
reaction and histamine would elicit automatieity in corresponding preparations of
the guinea pig.
METHODS
The experiments were performed on isolated
atria of adult guinea pigs of both sexes. The animals were previously sensitized with horse serum.*
Isolated Atria. The isolated atria were suspended in 50 ml. of oxygenated tyrode solution at
29 C. The left atrium was attached lower than
the right and was tied with a silk thread to a
glass hook. The upper part of the right atrium
then was joined to an isotonic lever which recorded
on the smoked paper of a kymograph.
In some experiments the right and left atria
were carefully isolated, placed in separate baths,
and attached to writing levers. The electrocardiogram was recorded with a model III D Grass eleetroeneephalograph. A unipolar lead was obtained
•Provided by the Iustituto Baeteriologico de Chile,
diluted with twice its volume of isotonie sodium
chloride solution, or with fresh egg white diluted
similarly. Sensitization was produced bj* 5 ml. of
diluted antigen intraperitoneally and 5 ml. intramuscularly, 50 to 60 days before the experiment.
by means of a U-shaped silver electrode in contact
with the central part of the left atrium, with an
indifferent electrode lying free in the bath. This
lead is similar to the one used by Garb and Chenoweth8 for recording the unipolar electrogram on
the isolated cat papillary muscle.
The anaphylactic reaction was elicited by the
addition of the specific antigen to the bath. In
other preparations obtained from normal guinea
pigs, the effect of various doses of histamine was
studied.
KESULTS
Effects of Histamine
The effects of different doses of histamine
on the isolated atria are summarized in
table 1. Changes in the rate and amplitude
of contraction of the preparation containing
both atria and of the isolated right atrium
are considered together, since in both cases
the spontaneous rhythm originated in the
sinus node. In view of individual variations,
the changes in amplitude are expressed in
per cent of the initial value.
Frequency and Rhythm. The addition of
different amounts of histamine to the bath
was followed by an increase in rate which
progressed with the concentration of the
drug between 1 X 10"4 mM/L. and 1 X 10""
niM/L., beyond which higher concentrations
elicited no further acceleration.
As a rule, lower concentrations of histamine (10-4 and 10"3 mM/L.) did not disturb
the rhythm, but higher concentrations (l(h 2
and 1(H) frequently produced irregularities
in the electrocardiogram unaccompanied by
changes in the shape of the complexes.
Amplitude of Contraction. The addition of
histamine induced an increase in the amplitude of contraction that appeared shortly
after the increase in frequency. The effect
on amplitude was also correlated with the
concentration up to 1 X 10~2 mM/L.
The increase in amplitude observed at the
latter concentration was 68.1 ± 15.9 per
cent of the control value. This increase was
not exceeded with higher histamine concentrations (table 1). An example of the effect
of various histamine concentrations appears
in figure 1.
Automaticity of Isolated Left Atrium.
The isolated left atrium bathed in tyrode
ANAPHYLACTIC REACTION
523
FIG. 1. Arrows, addition of histamine (£f) in various concentrations, 10 *, 10", 10~=, 10'1
mM/L., from left to right. Numbers, contractions per minute.
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solution at 29 C. usually does not exhibit
rhythmic aetivitj', either mechanical or electrical. An occasional spontaneous contraction
may be observed occurring at intervals
ranging from 0.5 to 3 min. Histamine
induced a regular automaticity in both electrogram and kymogram. Data summarized
in table 1 shows that the proportion of cases
in which histamine induced this effect increased with the concentration. In low concentrations ( I X 10"4 mM/L.) histamine did
not induce any automaticity while higher
concentrations succeeded in almost all cases.
Figure 2 shows an example of the automaticity induced by histamine. This effect generally appeared 2 to 3 min. after the addition
of the drug.
Dose-Response Relationship. The results
indicate that, with respect to effects on rate,
amplitude of contraction of both atria and
automaticity of the left atrium, a concentration of 10"4 mM/L. of histamine can be considered the threshold level and 10~2 mM/L.
the maximum. Concentrations ten times
higher than the latter had no greater effect.
Effect of the Anaphylactic Reaction
The results obtained in experiments with
specific antigens are summarized in table 1.
Frequency and Rhythm. The addition of 1
ml. of the antigen solution to the 50 ml.
bath containing the atria of a sensitized
guinea pig induced an increase in frequency.
The average increase observed in 12 preparations was 19 ± 4 per cent of the initial rate.
No disorders of rhythm were observed.
In similar experiments on 8 isolated atria
from normal nonsensitized guinea pigs, the
addition of the same amount of antigen had
no effect on frequency of contraction.
FIG. 2.—Isolated atria from the same normal guinea
pig in separate baths. Upper record, right atrium;
Lower record, left atrium. Time, 5 sec. intervals.
Arrow, addition of histamine (1 X 10'3 mM/L.).
Figures, contractions per minute.
Amplitude of Contraction. The addition of
the antigen caused the isolated atria of sensitized animals to show a constant increase in
amplitude of contraction. The average of 12
experiments was 40 ± 17.1 per cent of control values. In 8 isolated atria from normal,
nonsensitized animals the addition of the
antigen induced an insignificant decrease in
amplitude. Figure 3 shows the result of one
experiment in which the antigen was added
to the isolated atria of a normal and a sensitized animal.
Automaticity of Isolated Left Atrium. This
was studied in 8 isolated atria from sensitized
animals. The addition of 1 ml. of the antigen
PENNA, ILLANES, UBILLA, MUJICA
524
O,tM«
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FIG. 3.—Isolated atria from two different guinea
pigs. Upper record, atria of an animal sensitized to
horse serum; Lower record, from a normal. Arrow,
addition of 1 ml. of diluted horse scrum to the bath.
Figures, contractions per minute.
solution to the 50 ml. bath induced automatic! ty in 5 cases (fig. 4).
Delay of Ana,phyla.ctic Reaction. The data
summarized in table 2 show that the effect
of antigen on the rate and amplitude of contractions of the right atrium, or both atria appeared after a latent period ranging from 1
to 2 min. Automaticity of the left isolated
atrium appeared after a slightly greater delay (fig. 4).
Relationship Betiveen Effects of Antigen and
Histamine
The effects induced by the anaphylactic
reaction on the rate and amplitude of contraction are similar to those induced by histamine in concentrations between 1 X 10~3
and 1 X 10"2 m l l / L The proportion of
cases in which automaticity was induced by
the anaphylactic reaction in the isolated left
atrium was similar to that observed with hislamine at these concentrations. The anaphylactic reaction did not induce changes in
rhythm and these were seen after histamine
only in concentrations 1 X 10~2 mM/L. or
higher.
DISCUSSION*
The effects of histamine observed on the
preparation of both isolated atria of guinea
pigs are in agreement with those reported
by Biegler and Tiemann7 in the same prepara-
FIG. 4. Electrograms and kymograms of isolated
right and left atria from a sensitized guinea pig.
Upper records, left atrium; Lower records, right
atrium. A, 1 ml. of diluted egg albumin was added
to the bath. Figures, times at which the electrograms were taken. Vertical lines, 0.2 sec. intervals.
tion of the rabbit and by Burn9 in that of
the guinea pig. These experiments do not
reveal whether the effects on rate and amplitude of contraction are separable, or whether
the latter is merely secondary to the former,
as it has been shown to be in other isolated
heart preparations when stimulated electrically (Szent-Gyb'rgyi,10 Bowditch," Kruta,1Gold and Catell1*).
These experiments performed on the isolated left guinea pig atrium confirm in this
animal the automaticity-inducing effect of
histamine in the isolated left rabbit atrium
reported by Eiegler and Tiemann.7 The histamine concentrations used by these authors
were lower (about 1 X 10"3 mM/L., however,
than in this experiment and the frequency of
the automatic rhythm induced by histamine
was also lower.
The results reported here indicate that the
heart is a potential "shock organ" during the
anaphylactic reaction. The latent period of
about 1 min. observed in the anaphylaetic
reaction in the atria is in agreement with
AXAPHYLACTIC REACTION
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the observation of Schultz-Dale phenomenon
on the isolated uterus. The effects of the
anaphylaetic reaction are similar to those
induced by histamine and are quantitatively
equivalent to the effects of histamine in concentrations between 1 X 10"s and 1 X 10~2
mM/L. This relationship of concentrations
is similar to that observed by Varela14 in
the Schultz-Dale phenomenon on the isolated
guinea pig uterus.
These findings support the concept that
the cardiac effects of histamine and of the
anaphylaetic reaction are exerted directly on
the myocardium and are not dependent on
changes in coronary flow, since in the isolated atria preparation the coronary circulation is not involved and the antigen or drug
reaches the cell by diffusion from outside.
The fact that both histamine and the anaphylaetic reaction induce automaticity in the
isolated left atrium increases the evidence
supporting the similarity of histamine to the
substances liberated during the anaphylaetic
reaction.
SUMMARY
The comparative effects of histamine and
of the anaphylaetic reaction on the isolated
guinea pig atrium were studied. The dose:
response relationship for histamine showed
that, with regard to the increase of frequency
and amplitude of contraction as well as the
appearance of rhythmic activity in the left
isolated atrium, 1 X 10"4 mM/L. was a threshold concentration and 1 X 10"2 mM/L. was
maximal. In the isolated atria from sensitized guinea pigs, the addition of the specific
antigen induced effects to those of histamine in concentrations between 1 X 10"s and
1 X 10"2 mM/L., but only after a latent
period of about 1 min. Ilistamine and the
anaphylaetic reaction led to the appearance
of rhythmic activity in the isolated left
atrium. In both cases the automatic activity
began after a latent period of from 1 to 6
mins. No qualitative differences between
the effects of histamine and the anaphylaetic
reaction were observed. Since the latter effects
are due to sensitization of cardiac tissue
itself, the heart should be considered as a
525
TABLE 2.—Latent Period of Effect of Histamine and
Anaphylaetic Reaction on Isolated Guinea Pig Atria
Both Atria
and right atrium
Histamine
concentration
(mM/L.)
Number of
experiments
Effect of histamine
1 X 10"'
2-t
3
23
1 X 1025
1 X 10 =
26
1 X 10Effect of antigen
Sensitized
guinea pigs
12
Isolated
left atrium
Time Number of
range
experi(min.)
ments*
0
Time
range
(min.)
0
—
0
3
0
10
0
10
2 - 5
1 - 6
1 - 3
5
1, 8 - 2
1 - 2
*Onl3' cases exhibiting an effect considered.
potential shock organ during the anaphylaetic reaction.
SUMMARIO IN INTERL1NGUA
Esseva studiate le effectos comparative de
histamina e del reaction anaphylaetic super
le isolate auricula de porcos de India. Le
relation inter dosage e responsa pro histamina
monstrava que, con respecto al augmento del
frequentia e del amplitude de contraction e
etiam con respecto al apparition de activitate
rhythmic in le isolate auricula sinistre, le
concentration limine esseva 1 X 10"1 mM/L e
le concentration de efflcacia maximal 1 X
10-- mM/L. In auriculas isolate ab sensibilisate porcos de India, le addition del antigeno
specific induceva effectos simile a illos de
histamina in concentrationes inter 1 X 10~3
e 1 X 10"- mM/L, sed solmente post un per
iodo latente de circa un minuta. Histamina si
ben como le reaction anaphylaetic resnltava
in le apparition de un activitate rhythmic in
le isolate auricula sinistre. In ambe casos le
activitate automatic commenciava post un
periodo latente de inter un e sex minutas,
Xulle differentias qualitative inter le effectos
de histamina e le reaction anaphylaetic esseva observate. Yiste que le effectos hie discultite es causate per sensibilisation del
histos cardiac mesme, le corde debe esser
considerate como potentialmente un organo
de choc durante le reaction anaphylaetic.
PEXNA, ILLAXES, UBILLA, MUJTCA
526
Wirkung der "Substances actives." Pfliiger.
Arch. f. ges. Physiol. 222: 450, 1929.
REFERENCES
1.
J., AND ROBINSON, G. C.: An eleetrocardiographic study of the anaphylaetic
rabbit. J. Exper. Med. 18: 450, 1913.
AUER,
S. GARB, S., AND CHENOWETH, M. B.:
2. HECHT, A. F., ASD WENGRAF, F . : Elektro-
kardiographische Untersuchungen iiber Anaphylaktische Storungen der Herzehlage beiin
Kaninchen. Ztsehr. ges. Exper. Med. 2:
271, 1913-14.
3. CRIEP, L. H.: Electrocardiographic studies of
effect of anaphylaxis on cardiac mechanism.
Arch. Int. Med. 48: 1098, 1931.
4. WILCOX, EL, AND ANDRUS, E. C.: Anaphylaxis
in the isolated heart. J. Exper. Med. 67:
169, 1938.
Downloaded from http://circres.ahajournals.org/ by guest on June 11, 2017
5. ANDRUS, E. C, AND WILCOX, H.:
The effects
of anaphylaxis and of histamine upon the
coronary arteries in the isolated heart. J.
Exper. Med. 69: 545, 1939.
fi. KEM-NER, A., PEXNA, M., AND SCHWEID, A.:
Cardiac anaphylaxis. Circulation 12: 730,
]955.
7. RIGLER, R., AND TIEIIANN, F . : Fortgesetzte
Untersuchungen iiber das Hcrzhorinon. Die
9.
10.
11.
12.
The T
deflection of isolated mammalian heart
muscle electrograii). Circulation Research
1: 135, 1953.
BURN, J. H.: Practical Pharmacology. Oxford
Blackwell Scientific Publication, 1952, pp.
23-24.
SZEXT-GYORGI'I, A.: Chemical Physiology of
contraction in Body and Heart Muscle.
New York, Academic Press, 1953, pp. 71-73.
BOWDITCH, II. P.: Cited by (10), p. 71.
KRUTA, V.: Sur F activite rythmique du muscle cardiaque; variations de la reponse mecanique en fonction du rythme. Arch.
Internat. physio]. 45: 332, 1937.
13. GOLD, H., AND CATTETJIJ, M.: Mechanism of
digitalis action in abolishing heart failure.
Arch. Int. Med. 55: 263, 1940.
14. VARELA, A.: Estudio sobre la accion de algunos inhibidores de la flbra muscular lisa
en el Fenomeno de Sehultz Dale. Tesis de
Licenciatura, Facultad de Medicina, Univ.
de Chile. Santiago, Chile, Stanley, 1944.
Effect of Histamine and of the Anaphylactic Reaction on Isolated Guinea Pig Atria
MARIO PENNA, ALEJANDRO ILLANES, MARIO UBILLA and SERGIO MUJICA
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Circ Res. 1959;7:521-526
doi: 10.1161/01.RES.7.4.521
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