Effect of Tourniquet Shock and Acute
Hemorrhage on the Circulation of
Various Organs in the Rat
By Louis Takdes, M.D., Kdlmdn Kdllay, M.D., and
Josephine H. Skolnik, Ing. Chem.
With the technical assistante of V. Vajda, A. Turcsdnyi, K. Albert, and A. Karal
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• Changes in cardiac output and the circulatory responses of certain organs to various
forms of stagnant hypoxia (hemorrhage,
shock, dehydration, etc.) have been studied
by many investigators, including "Wiggers,1
Gomori and Takacs,2 and Sapirstein, Sapirstein, and Bredemeyer.3 It has been gradually
accepted that blood flow in various organs
does not change uniformly; compared to the
limited change in coronary and cerebral blood
flow, the renal blood flow is severely affected.
In earlier experiments, Gomori et al., Takacs,
and Kail ay have studied the cardiac output
and circulatory response of the heart,4 kidneys,5"8 and of the extremities,9 in shock,
dehydration, and arterial hypoxia in dogs.
Having observed a similar shifting in the
distribution of cardiac output (increase of the
coronary fraction and decrease of the fractions of the kidneys and of the extremities),
both in "hypovolemic" hypoxia (low cardiac
output) and in "hypervolemic" (arterial)
hypoxia with high cardiac output, we have
assumed that the same factor, namely, hypoxia of the tissues, may be responsible for
the changes involved.
Experimental methods used in the past
were not suited for measuring the circulation
of several organs simultaneously. In the
above-mentioned states, most authors measured only the blood flow and vascular resistance of the organs and neglected the simultaneous determination of cardiac output.
Thus, they failed to point out the changes in
the distribution of reduced cardiac output.
From the Second Department of Medicine (Head:
Professor P. Gomori), University Medical School,
Budapest, Hungary.
Received for publication August 30, 1961.
Circulation Research, Volume X, May 1962
Some years ago, Sapirstein10'11 described
an "indicator fractionation" method using
first K42 and later Rb86. This method offers
a possibility of measuring simultaneously the
fractionation of cardiac output among all
organs except the brain,12 because the K4~
and Rb86 extraction ratios of all organs other
than the brain are virtually the same as those
of the whole body during the period of the
experiment. As the isotope injected is diluted
in the total cardiac output, the quantity of
the isotope accumulating in the various organs is proportional to their fraction of the
cardiac output. Determination of cardiac output and blood pressure then makes possible
the calculation of the blood flow and the vascular resistance of the organs.
Sapirstein investigated with K42 the circulation of rats after "mild" (10 ml./Kg.) and
"moderate" (21 to 25 ml./Kg.) bleeding.13
After repeating Sapirstein's experiments with
Rb86, we studied the effects of more severe
hemorrhage and of tourniquet shock on the
circulation in rats.14 Regarding the effect of
moderate hemorrhage, our results with K 4213
and Rb8e 3 show close agreement with his.
Methods
Essentially, we followed Sapirstain's method.11
Seventy-one male rats, weighing 140 to 280 Gin.
and fasted for 18 to 24 hours, were used. The
animals were anesthetized with 40 mg./Ivg.
sodium pentobarbital intraperitoneally (I.P.), »s
we had found that this drug is better suited for
the actual purpose in the rat than urethane or
chloralose.15
Tourniquet shock was produced by ligation of
both hind legs with a rubber band for three hours
in unanesthetized rats. Circulatory studies were
begun one to two-and-a-half hours after the removal of the ligature and after giving sodmmpentobarbital I.P.
753
754
TAKACS, KALLAY, SKOLNIK
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Two bleeding' groups were set tip: from one
group tin average of 23 (20 to 26 ml./Kg-.) nnd
from the other an average of 32 ml./Kg. (27 to
39 ml./Kg.) blood were withdrawn through a
catheter in the ciirotid artery. Measurements were
taken 5 to 20 minutes after hemorrhage.
Blood pressure wns measured in the carotid
tirtery with membrane manometer, Vorsatz-Kiillay.
Coagulation of the blood was prevented by injocting 2.5 nig1, heparin in 0.2 ml. of saline into
the tail vein. Cardiac output was determined
with Evans blue dilution10: 0.3 ml. 1.5 per cent
dye solution was injected into the femoral vein and
blood samples were taken from the carotid artery
with the Krillay-Vorsntz type fractionating blood
collector at 0.66-second intervals from the controls and at one- and two-second intervals from
the shocked and bled groups, respectively. Dye
concentration was measured in 20 /U. blood diluted
with physiological saline to 3.0 ml. A Beckman
B. spectrophotometer, set at 590 m/.i, was used.
Tbe cardiac output fractions of the various
organs were determined by the injection of 5 to
10 ixc. Rbs« Cl in 0.5 ml. physiological NnCl solution into the femoral vein. The animals were
sacrificed after 70 to 80 seconds (those subjected
to severe hemorrhage after 120 seconds) by injecting1 0.5 ml. saturated KC1 into the tail vein.
The experiment thus proceeded as follows: anesthetization, measuring of blood pi'essure, administration of Rbso, and after 55 to GO seconds
determination of cardiac output, sacrifice. The
organs were then dissolved in a 20 per cent KOI1
solution, and the radioactivity of the samples was
measured with a Geiger-Miiller tube.
Having thus obtained values for the cardiac
output and measured tbe weight of the organs,
we calculated the blood flow of each organ. Circulatory resistance of the whole body and of the
organs were calculated from the cardiac output
or blood flow and arterial blood pressure (taking
venous pressures as 0) nnd were expressed in
dynes/sec./cnr-"'. Flow and resistance values were
calculated for ]00-Gm. organ weight.
STATISTICAL ANALYSIS
As the within-sample variances proved to be
practically equal (Bartlett's test), the standard
deviations were computed from the pooled sum-ofsquares. Although in a few cases this pooling was
not entirely legitimate, it was used in order to
compile a, uniform and synoptical table, as it did
not materially affect the results. The data were
tested by the Student's t.
Results and Discussion
The results are summarized in table 1.
The average cardiac output of control animals was 28.6 ml./lOO Gm., which is somewhat
higher than Sapirstein's17 value (23.1). The
mean value of blood pressure was 120 mm.
Hg; mean resistance was 336 X 103 dynes
/sec./cnr 5 .
The mean value of cardiac output in tourniquet shock Mras 10.1 ml./lOO Gm., while
blood pressure decreased to an average level
of 50 mm. Hg. The corresponding mean resistance was 407 X 103 dynes/sec./cm~r>.
The mean value of cardiac output in the
moderately bled group (23 ml./Kg.) was very
low, not more than 6.0 ml ./100 Gm. with a
blood pressure of 47 mm. Hg, while resistance
rose to 709 X 103 dynes/see./enrr>. After more
severe hemorrhage (32 ml./Kg.) mean blood
pressure was only 18 mm. Hg.
TOURNIQUET SHOCK
The blood flow showed the following pattern : there was a reduced blood flow in all
of the organs. The smallest decrease was
found in the heart, i.e., to 58 per cent of the
control; the greatest changes were found in
the kidney (to 26 per cent) and in the skin
(to 23 per cent). While in the controls, renal
blood supply was nearly twice the coronary
(calculated for 100 Gm. weight of organ) ;
in shock, coronary blood flow was greater than
renal.
, ,
There was a significant increase in the vascular resistance of the skin and of the "carcass" (skeleton, muscles, and endocrine
organs). Due to the great standard deviation,
there was no change in the blood flow of the
other organs which •would be statistically
significant, although,the differences between
some of the means were fairly large. Thus
there was an average increase in renal resistance from 29 to 71 (the difference is near the
significance level, P—0.05), while the circulatory resistance of the intestines increased
from 136 to 183. The resistance of the
"lungs" (bronchial circulation) showed a
minor but significant decrease.
Considerable changes •were observed in the
distribution of the cardiac output: the coronai-y fraction increased definitely from 2.8
to 4.7 per cent, and the fractions in the
"lungs" and liver to a lesser degree. The
Circulation Research, Volume X, May 1062
SHOCK AND HEMORRHAGE
755
TABLE 1
Effect of Tourniquet-Shock or Hemorrhage on the Circulation in the Hat
Number of cases
Control
Tourniquetshock
22-30
19-20
Hemorrhage
ml./Kg.
ml./KB.
23
32
10-13
g
Within-sample
standard
deviation
Mean values
Total body:
Cardiac output
Blood pressure
Resistance
Heart:
Jilood flow
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Resistance
fraction
Kidney:
Blood flow
Resistance
Fraction
Lung:
Blood flow
Resistance
Fraction
10.lt
28.6
120.0
336.1
50.5}
407.0
6.0}
47.3}
708.6}
2 l I .3
50.9
2.8
122.0}
41 .6
4.7}
74.8}
61.9
4.6}
415.0
29.0
14.5
107.2}
70.8
9.8}
50.1}
106.2t
6.4}
97.7
124.5
49.6}
95.6*
4.11
27.2}
156.5
4.8}
3.1.4}
8.6
189.4
11.31
13.4}
344.81
7.0
75.8
135.8
17.0
34.8}
182.2
19.1
15.0}
299.01
1.5.2
14.6
3.9
25.6
30.4t
oo o
21.4
5.8
12.0
964.6
8.8
2.9}
2423.3*
5.6}
19.6
526.8
45.8
6.5}
786.2t
45.2
2.9
18.0}
6.6
17.6
219.9
10.0}
74.2
18.6
1.0
5.7}
113.5
70.9
3.3
7.1}
4.4.7
1.1
32.3
T-jiver:
Blood flow
Resistance
Fraction
Intestine:
Blood flow
Resistance
Fraction
Splanchnic bed
liver and intestine:
Fraction
Skin:
Blood flow
Resistance
Fraction
Carcass:
Blood flow
Resistance
Fraction
Scales:
67.2
] 70.4
19.9
140.2
6.8*
3.1
27.8
119.2
1.6}
3.9
3496.lt
2120.3
5.4}
2.5}
2.5
4.9}
4.6
887.7t
55.1}
299.9
53.3}
6.4
Cardiac output—nil./niin./lOO Gin. body weight
Blood pressure—nun. Hg
Resistance (body)—.10" dynes sec. cnr'/lOO Gin. body weight
Blood flow (organ)—nil./niin./JOO Gm. weight of orgai l
Resistance (organ)—10"' dynes sec . cnf'/lOO Gm. weight of organ
Fraction—blood flow of total organ expressed in per cent of total cardiac output
The mean values from experimental groups were compared with the mean values of the
control group. The differences were statistically evaluated with the following results:
*P < 0.05.
XT < 0.01.
}P < 0.001.
No symbol, V > 0.05.
Circulation lirsearak. Volume X, May 11)02
756
fraction in the spanchnic bed (liver + intestines) rose from 25.6 per cent to 30.4 per
cent. A significant decrease occurred in the
fractions distributed in the kidney and the
skin.
HEMORRHAGE
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As cardiac output was not measured in
more severe hemorrhage, blood flow and vascular resistance in hemorrhage were calculated in the moderately bled groups only. The
blood flow of all organs decreased considerably. Coronary circulation suffered the least;
nevertheless it reached only 35 per cent of
the controls. Eenal and cutaneous circulations were the most severely affected, the blood
flow being not more than 12 to 13 per cent
of the controls. The vascular resistance of the
organs except coronary and bronchial vessels
increased significantly.
Compared to the controls, the distribution
of cardiac output showed a considerable
change both in moderate and severe hemorrhage. Changes were in the same direction in
both groups, although generally greater in
severe hemorrhage. Increases in the fractions
of the heart and the "lungs" were considerable and approximately of the same order.
At both hemorrhage levels, the fraction of
the "carcass" was significantly greater, i.e.,
55.1 and 53.3 per cent, respectively, than the
45.8 per eent of the controls. Eenal and cutaneous fractions showed considerably lower
values. Fractions of the liver and intestines
were somewhat lower in both groups, but this
change only becomes significant in severe
hemorrhage for the liver fraction. The total
splanchnic fraction of the cardiac output
dropped in hemorrhage from 25.6 per cent to
22.2 and 21.4 per cent, respectively. These
changes considered separately were not significant, but when the two groups were
pooled they reached the limit of significance
(P < 0.05).
Eegarding the moderately bled group, our
data are in complete agreement with Sapirstein's results.3
Thus, we found in the rat some essentially
common circulatory features, developing in
20 minutes after hemorrhage and which ap-
TAKACS, KALLAY, SKOLNIK
pear a few hours after the ligation of the
extremities (tourniquet shock). In both cases,
blood flow in all organs decreases and
vasoconstriction appears in all the organs
observed except the heart and the "lung." Increase in the coronary fraction of cardiac output and decrease of the renal and cutaneous
fractions represent a further common characteristic feature. Similar changes in renal
circulation in hemorrhage, tourniquet shock,
and in dehydration have been demonstrated
by a corrosion method in dogs.18
In these states, there is a circulatory shifting by which the myocardium is favored at
the expense of the kidney and the skin. The
basic facts of circulatory redistribution observed in the rat are similar to those which
others and we ourselves have earlier observed
in dogs and men.
In tourniquet shock and moderate hemorrhage, blood pressure was more or less the
same, but there was a more pronounced decrease in cardiac output and in the blood
flow of all organs after hemorrhage, that is
to say, hemorrhage caused a greater rise in
vascular resistance. We did not study the
cause of this difference, but it may be assumed
that at an identical reduction in blood pressure there is a greater decrease in the circulating blood volume after hemorrhage.
It is rather surprising that in both bleeding
groups we found increased "carcass" (bone
and muscle) fractions. Sapirstein et al.3 reports the same observation after moderate
hemorrhage. The reason for this phenomenon
is not yet clear.
Summary
Cardiac output, blood pressure, and organ
fractions of cardiac output, except the brain,
were determined in rats subjected to tourniquet shock and to hemorrhage. Bleeding involved the withdrawal of 23 and 32 ml./Kg.
blood, respectively, and determinations were
carried out 5 to 20 minutes after bleeding.
Sapirstein's'' indicator fractionation'' method
with Eb86 was used for the determination of
the cardiac output fractions. Blood flow and
circulatory resistance of the organs were calculated from the experimental data. Cardiac
Circulation Resoarch, Volume X, May 1962
757
SHOCK AND HEMORRHAGE
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output was not measured after severe (32
ml./Kg.) hemorrhage. In tourniquet shock at
50 mm. Hg blood pressure, the cardiac output
and the blood flow in all organs decrease
while vascular resistance of the skin and of
the "carcass" (skeleton, muscles, endocrine
organs) increases. The heart, "lung," and
the liver fractions of cardiac output increase,
renal and skin fractions decrease. After moderate hemorrhage at 47 mm. Hg blood pressure, cardiac output and the blood flow of
the organs are low. Vascular resistance of all
organs other than the heart and the "lung"
rises. Cardiac output fractions of the heart,
"lung," and "carcass" increase; renal and
cutaneous fractions decrease. In severe hemorrhage, the redistribution of cardiac output
lias a similar tendency but is usually more
pronounced.
W.: Control of venal circulation in hypoxia.
Acta med. Acad. sci. hung. 16: 43, 1960.
7. TAKACS, L., AND KALLAY, K.: Changes in renal
circulation in exsiccosis. Acta med. Acad. sci.
hung. 10: 309, 1957.
8. TAKACS, L., AND KALLAY, K.: Renal circulation
in traumatic shock. Acta physiol. Acad. Sci.
hung. 12: 373, 1957.
9. TAKACS, L.: Circulation of limbs in hypoxic
conditions. Acta physiol. Acad. sci. hung. 11:
189, 1957.
10. SAPIRSTEIN, L. A.: Fractionation of the cardiac
output of rats with isotopic potassium. Circulation Eesearch 4: 689, 1956.
11. SAPIRSTEIN, L. A.: Regional blood flow by
fractional distribution of indicators. Am. J.
Physiol. 193: 161, 1958.
12. SAPIRSTEIN, L. A., AND HANUSEK, G. E.: Cere-
bral blood flow in the rat. Am. J. Physiol.
193: 272, 1958.
13. SAPIRSTEIN, L. A.: Effect of hemorrhage on distribution of cardiac output. Fed. Proc. 16:
111, 1957.
14. TAKACS, L.,
Acknowledgment
We wish to express our thanks to Dr. Ireneus
Juvancz and Mrs. J6zsef Tamassy (of the Biomotric Department of the Mathematical Research
Institute, Hungarian Academy of Science) for having kindly carried out the statistical calculations.
KALLAY, K.,
AND SKOLNIK,
J.:
Studies on the renal, cardiac and skin fraction of the cardiac output in rats with Rb80
in ischemic shock and hemorrhage (preliminary communication)- Acta med. Acad. sei.
hung. 14: 457, 1959.
15. KALLAY, K-, AND TAKACS, L.: Study of various
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Effect of Tourniquet Shock and Acute Hemorrhage on the Circulation of Various Organs in
the Rat
Louis Takács, Kálmán Kállay and Josephine H. Skolnik, Ing. Chem
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Circ Res. 1962;10:753-757
doi: 10.1161/01.RES.10.5.753
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