Renal Vein Renin and Juxtaglomerular
Activity in Sodium-Depleted Subjects
By Horacio Ajxen, M.D., John L. Simmons, M.D., and
James W. Woods, M.D.
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• In both man and experimental animals,
a low sodium intake or sodium depletion
increases aldosterone excretion1 and secretion.2 Observations on dogs with hyperaldosteronism secondary to chronic sodium depletion revealed that aldosterone secretion
decreased markedly following nephrectomy.:!
The large body of evidence showing that the
aldosterone-stimulating hormone arises from
the kidney and is renin has recently been summarized by Davis.4 Measurements show an
elevated peripheral plasma level of renin in
normal human subjects on a low sodium intake5"7 or depleted of sodium.7 Goormaghtigh's hypothesis8 that juxtaglomerular (JG)
cells secrete renin has recently been strengthened by: correlations between renin content
and degree of granulation of JG cells under
different experimental conditions in rats, 9 ' 10
microdissection studies,11 and application of
the fluorescent antibody technique.12 In a
postmortem study, the degree of granulation
of the juxtaglomerular apparatus was found
to be correlated inversely with levels of plasma sodium present during the week prior
to death.13
Since correlation of renal vein renin and
JG granularity in human subjects depleted
of sodium has not been reported, the present
study was designed to accomplish this.
Methods
Ten patients who required surgery for renal
From the Departments of Medicine and Urology,
University of North Carolina School of Medicine,
Chapel Hill, North Carolina.
Supported by U. S. Public Health Service Grant
HE-03794 from the National Heart Institute and
Grant FR-46 from the General Clinical Research
Centers Branch of the Division of Research Facilities
and Resources.
Accepted for publication January 12, 1965.
130
calculi or cysts were admitted to the hospital
and were, as volunteers, used for special study.
None had hypertension, nephrosis, cirrhosis of the
liver or heart failure and all were normally hydrated. Age and sex are shown in table 1. Eight
of ten patients required pyelolithotomy, one had
a renal cyst excised (tumor could not be excluded), and one, in whom renal tumor was suspected, was found to have a normal kidney at
exploration. Base line studies, in addition to history, physical examination, hemogram, and urinalysis, included serum electrolytes, fasting blood
sugar, bromsulphalein test, endogenous creatinine clearance,14 blood urea nitrogen, urinary sodium, urinary potassium, and bio-assay for renin
(Helmer aortic strip method) 15 in peripheral venous blood. Five patients ate a regular diet and
served as controls. Five patients ate a diet containing 87 mEq of sodium, 75 mEq of potassium with a 40 mEq potassium supplement and
received 1 g of chlorothiazide daily for ten
days. More drastic sodium restriction was not
instituted because of the impending surgery. Urinary sodium and potassium were measured daily
and serum electrolytes every third day.
Anesthesia was induced by thiopental sodium
and nitrous oxide, and was maintained by means
of nitrous oxide and halothane. At operation,
samples of peripheral and renal venous blood
were obtained for renin bio-assay. An open renal
biopsy was taken and an intravenous infusion of
physiological saline solution started immediately
thereafter. Specimens were always taken shortly
after exposure of the kidney and prior to renal
surgery. No operative or postoperative complications occurred.
Renal biopsy material was fixed in Helley's
solution and stained with the Bowie stain for
determination of the "juxtaglomerular index"
(JGJ). 16 The JGI was determined without knowledge of the biopsy's origin by one of the investigators (HA). Blood specimens for bio-assay
were collected in heparinized syringes and immediately placed in ice. They were then centrifuged, the plasma removed, the latter acidified
to pH 5.5 with 0.1 N hydrochloric acid, and
frozen for subsequent bio-assay on the rabbit aortic strip.
Circulation Research, Vol. XVII, August 1965
RENIN AND JG ACTIVITY IN SODIUM DEPLETION
131
TABLE 1
Renal Vein Renin and Juxtaglomerular Index in Sodium-depleted and Control Subjects
Subject
no.
1
2
3
9
10
Age
Sex
Sodium-depleted
Renin
ng/ml*
51 F
60 F
53 F
24 F
46 M
30
62
37
40
80
Controls
Juxtaglomerular
index
Subject
no.
Age
Sex
Renin
ng/ml
Juxtaglomerular
index
12.9
25.8
25.4
19.0
17.6
4
58 F
51 F
58 F
38 M
64 F
0
0
0
0
0
3.1
4.0
—
2.2
5.5
5
6
7
8
*Renin is reported as nanograms of angiotensin n formed per ml plasma.
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5
DAYS
6
FIGURE 1
Mean daily sodium excretion of five subjects receiving the sodium-depletion regimen. Vertical broken
lines indicate range of excretion. C (at left) represents
control day prior to institution of the regimen.
Results
Sodium balance data for the five experimental subjects are shown in figure 1 and
table 2. Markedly negative sodium balance
was present during the first two days of the
regimen and net balance for the experimental period was negative in three of the subjects. Two subjects showed net positive balance during the period by reason of reduced
urinary sodium excretion. Body weight of
each subject decreased approximately 2 kg
during the first three days and thereafter remained stable. Serum sodium decreased an
average 4.0 mEq/liter (range 2 to 5.5). Potassium intake was 115 mEq/day and depletion did not occur. Blood urea nitrogen
rose slightly and creatinine clearance decreased in three of the subjects while on the
regimen. These values returned to normal
during the immediate postoperative period.
Circulation Research, Vol. XVII, August 196)
Results of the renin bio-assays and JG index
determinations are shown in table 1. Renal
vein renin was significantly elevated in all of
the subjects on the sodium-depletion regimen
as compared with controls, none of whom
had measurable renin (P < 0.01). Calculation of the JG index was not possible in one
control subject due to.an inadequate biopsy,
but the difference between experimental and
control subjects was also statistically significant (P < 0.02) with the former showing
hypergranularity. An example (subject 2)
of a bio-assay record is shown in figure 2.
An example of a JG apparatus from a control
individual (subject 4) is shown in figure 3
and from a sodium-depleted individual (subject 2) in figure 4. Tests for renin in peripheral
FIGURE 2
Effect of 1 vd plasma samples and standards on spirally cut strip of rabbit aorta. 1: 0.1 fig synthetic
angiotensin 11 (standard). 2: peripheral venous plasma
of a patient with renal artery stenosis. 3,4: peripheral
venous plasmas of two patients with essential hypertension. 5: 0.1 tig synthetic angiotensin II. 6: peripheral venous plasma of subject number 2 prior to
sodium-depletion regimen. 7: right renal vein plasma
of subject number 2 after 10 days on an 87 mEq
sodium diet and 1 g of chlorothiazide daily. 8: peripheral venous plasma at the same time. 9: 0.1 ng
synthetic angiotensin II.
132
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Section of kidney from subject number 4. Lower half
of a glomerulus (Clom.) occupies the upper half of
the photomicrograph. Glomerular vascular pole is on
the left, the macula densa, lower right, and a tangentialhj cut arteriole, on the right. Note the absence of
juxtaglomerular granules and of cellular hyperplasia
(JG index = 3.1). Bowie stain.
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FIGURE 4
Section of kidney from subject number 2. The lower
margin of a glomerulus (Glom.) is in the upper center,
the glomerular vascular pole occupies the greater part
of the photomicrograph and a small section of an
arteriole (Art.) is at the upper right. Hypergranularity
and hyperplasia of JG apparatus are present (JG index = 25.8). Bowie stain.
Circulation Research, Vol. XVII, August 1965
133
RENIN AND JG ACTIVITY IN SODIUM DEPLETION
venous plasma were negative in all ten subjects at the beginning of the study and were
positive in two of five experimental subjects
and in none of the controls at the end of ten
days.
Discussion
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The role of the juxtaglomerular apparatus
as a volume receptor is supported by data
from many laboratories and results described here parallel those previously reported
from animal experiments. It is interesting
that hyperplasia and hypergranularity of the
JG apparatus and increased renal vein renin
were present despite only moderate sodium
depletion and also present in two subjects
who showed net positive sodium balance at
the time the specimens were obtained. Net
positive sodium balance in subjects 1 and 9
may be more apparent than real, however,
since the sodium in rejected food was not
measured, and their intake of sodium may
have been less than the 87 mEq allowed.
Body weight measurements suggest that
positive sodium balance did not occur. In
this study, it is not possible to discriminate between reduced plasma volume and decreased
total body sodium content as the continuing
stimulus for renin secretion. Extrarenal sodium loss was not calculated during this
study, but it is assumed that these figures
would not have altered significantly the sodium balance data. Creatininc clearance decreased in three of the five patients (numbers 2,3,10). Reduced glomerular filtration
with a decrease of filtered sodium is an additional explanation for decreased urinary excretion of sodium. However, net sodium balance was negative in the subjects whose
creatinine clearance became less; whereas
in the two with no significant change in
creatinine clearance, sodium balance was
positive.
As pointed out by several workers previously, comparison of results of renin bio-assay
from different laboratories is made difficult
by the large number of bio-assay methods
in use, as well as by the lack of an international standard for angiotensin. The
aortic strip method as used in our laborCirculation Reiearch, Vol. XVII, August 796.5
atory yields consistently a 4 to 4.5 cm contraction in response to 0.1 fig of synthetic
angiotensin (l-L-asparaginyl-5-L-valyl angiotensin octapeptide-Ciba). In sensitive preparations, as little as 0.005 fig of angiotensin,
added to 20 ml of modified Krebs solution
in the muscle chamber, can be detected.
Helmer has shown that the method is specific
for renin.15 We have not been able to demonstrate renin in the peripheral venous plasma
of patients with acute glomerulonephritis or
coarctation of the aorta. We have found renin
rarely in normal subjects and in those with
benign essential hypertension, but we have
found large amounts in the majority of patients with malignant hypertension. Renin
has been found in the peripheral venous
blood of only three of sixteen patients with
hypertension associated with renal artery
stenosis. It was present in the renal venous
blood in five of these sixteen individuals. That
increased renin activity in peripheral venous
blood could be demonstrated in only two of
five sodium-depleted subjects is in keeping
with the sensitivity of the method as performed in our laboratory. Others5'Oi 7 using
different bio-assay methods have consistently
demonstrated renin in the peripheral blood
of sodium-depleted subjects.
Summary
Ten patients requiring surgery for renal
cysts or calculi and having no evidence of
secondary hyperaldosteronism were divided,
as volunteers for special study, into experimental and control groups. After base line
studies, the experimental group was given a
diet containing 87 mEq of sodium and 1.0 g
of chlorothiazide daily for ten days. Control
subjects ate a normal diet. At operation,
samples of peripheral and renal venous blood
were obtained for measurement of renin activity and renal biopsy was taken for determination of the juxtaglomerular index.
Increased renin in renal venous plasma and
hypergranularity of the juxtaglomerular apparatus were found in sodium-depleted subjects but not in control subjects. These findings confirm results previously found in
animals.
134
AJZEN, SIMMONS, WOODS
genie de l'hypertension arterielle. Rev. Beige
Sci. Med. 16: 65, 1945.
Acknowledgment
We are indebted to Dr. Jack Hughes for his generous help in the procurement of suitable subjects
and to Miss Billie S. Bush for valuable technical
assistance.
References
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Circulation Research, Vol. XVII. August 1965
Renal Vein Renin and Juxtaglomerular Activity in Sodium-Depleted Subjects
HORACIO AJZEN, JOHN L. SIMMONS and JAMES W. WOODS
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Circ Res. 1965;17:130-134
doi: 10.1161/01.RES.17.2.130
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