(CANCER RESEARCH 39, 1269-1278, April 1979]
0008-5472/79/0039-0000$02.00
Effects of Mannitol or Furosemide Diuresis on the Nephrotoxicity and
Physiological Dispositionof cis-Dichlorodiammineplatinum-(ll)
in Rats1
Martin F. Pera, Jr.,2 Bernard C. Zook, and Harold C. Harder3
Departments of Pharmacology (M. F. P., H. C. H.J and Pathology (B. C. Z.J, The George Washington University Medical Center, Washington, D.C. 20037,
and Departmentof Pharmacology,OralRobertsUniversity,Tulsa,Oklahoma,74171(H. C. HJ
ABSTRACT
Although furosemide and mannitol have been reported to
protect against cis-dichlorodiammineplatinum(ll)
(COOP)induced nephrotoxicity, the possibility that these diuretics
might act in different ways to alter the physiological dispo
sition) and in vivo antitumor activity of COOP has not been
adequately assessed. Therefore, we compared the effects
of furosemide (12.5 mg/kg i.p. with 6.0 ml 0.9% NaCI
solution) and mannitol (300 mg in 3.0 ml 0.45% NaCI
solution as a 30-mm i.v. infusion) on the nephrotoxicity and
physiological disposition of COOP (6 mg/kg i.v.) in male
F344 rats. Serial histopathological evaluation of kidneys
indicated that an approximately equivalent degree of proxi
mal tubular necrosis occurred in rats given COOP alone or
with either furosemide or mannitol 1 to 4 days after drug
administration. Thereafter, a trend developed toward less
persistence of damage in the mannitol groups and progres
sion of tubular injury in furosemide-treated rats compared
to COOP alone (5 to 10 days after drug administration).
However, renal function, assessed by measurement of
blood urea nitrogen, estimation of glomerular filtration rate,
and p-aminohippurate clearance was partially protected in
rats given either diuretic.
The administration of furosemide or mannitol did not
markedly affect the triphasic plasma decay of platinum. A
similar uptake of platinum into spleen and small intestine
was seen at early time points (2 mm to 2 hr) in groups given
COOP alone or with either diuretic. Kidney platinum content
in rats receiving mannitol was similar to that of animals
receiving COOP alone. Kidneys of furosemide-treated ani
mals contained higher levels of platinum than did kidneys
of rats given COOP alone or with mannitol at 1, 2, 24, and
96 hr after treatment. Total 24-hr urinary excretion of
platinum was decreased, although not significantly, by both
diuretics. Furosemide or mannitol diuresis resulted in a
significant decrease in platinum concentration in 0- to 24hr urine samples. Thus, under these conditions the partial
protection of renal function observed with diuretics is not
the result of increased excretion of platinum in urine, faster
plasma clearance of platinum, or decreased renal levels of
platinum. However, a mannitol-induced diuresis may re
duce the duration of tubular necroses. It is possible that'
while tubular necrosis might be related to cumulative plati
num uptake in the kidney, the reduction of platinum con
centration in the tubular fluid caused by the diuretics might
account for the protection of renal function.
INTRODUCTION
COOP4 is an inorganic coordination complex which dis
plays activity against a variety of experimental and human
neoplasms (5, 14, 15, 25). However, administration of the
compound at therapeutic doses produces renal toxicity in
rats, dogs, monkeys, and humans (8, 12, 14, 26, 32).
Clinical interest in CODP has increased considerably since
the publication of several reports indicating that induction
of diuresis prior to COOP administration to cancer patients
ameliorated the renal damage caused by the drug (3, 13,
20). However, the possibility that various diuretics might act
in different
ways to alter
the physiological
disposition,
toxicities, and the in vivo antitumor action of CDDP has not
been adequately assessed. Therefore, we have undertaken
to investigate the toxicological, pharmacokinetic, and ther
apeutic aspects of the interaction between COOP and di
uretic drugs. In this paper, we evaluate the effects of
furosemide- and mannitol-induced diuresis on CDDP renal
toxicity and physiological disposition in rats. In an accom
panying report (23), we describe effects of diuretics on
acute lethality, gastrointestinal and hematopoietic toxicity,
and antitumor activity of COOP.
Cvitkovic et a!. (7) found that coadministration of COOP
with mannitol or large doses of i.v. fluids prevented the rise
in BUN and serum creatinine which occurs following COOP
administration at toxic doses in dogs. These investigators
also presented some data to suggest that the level of
platinum in serum after COOP was similar in dogs given
mannitol or prehydration. The data provided no comparison
with dogs given COOP alone. The same study stated that
the mannitol and prehydration treatments did not alter total
urinary clearance of platinum, although apparent increases
in the
recovery
of platinum
in urine
were
reported
in
prehydration and mannitol groups.
Ward et a!. showed that the male F344 rat is sensitive to
Grant CA-02978from the NationalCancerInstitute, Departmentof Health,
Education,and Welfare.A portion of this work was presentedpreviouslyin the nephrotoxic effects of CODP (32) and that renal dam
abstractform (24).
age, evidenced by elevation of BUN and necrosis of cells in
the
proximal tubule, could be ameliorated by administration
Sciences, The George Washington University, in partial fulfillment of the
I
Supported
2 From
by
a
dissertation
Grant
Cl-107
to
be
from
the
presented
American
to
the
Cancer
Graduate
Society
School
and
of
Arts
by
and
requirements for the Ph.D. degree.
3 To
whom
all
correspondence
should
be
addressed
University. Tulsa, Okla. 74171.
ReceivedJuly 24, 1978;acceptedJanuary3, 1979.
at
Oral
Roberts
4 The
abbreviations
used
are:
CDDP,
cis-dichlorodiammineplatinum(lI);
BUN,blood urea nitrogen; DTPA,diethylenetriaminepentaacetate;
PAH,p
aminohippurate; [3HJPAH,p-[glycyI-2-3H@aminohippuric acid.
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1269
M. F. Pera, Jr. et a!.
of furosemide prior to the platinum drug on an acute or
chronic basis (33, 34). In contrast to the results of Cvitkovic
5 to 6 weeks of age, the animals were separated into 4
treatment groups. The first group received 0.5 ml of 0.9%
et al. (7)with mannitolindogs,the reportof Wardet al. (34) NaCI solution i.v. All i.v. injections were made into the
mentioned unpublished work indicating that furosemide
lateral tail vein using a 26-gauge x 0.5-inch needle. The
treatmentof ratsresulted
inincreasedlevels
of platinumin second group received COOP (6 mg/kg i.v.), the highest
blood and kidney. We therefore extended the studies of dose of COOP that can be administered to these rats
these groups to provide a more detailed comparison of the without producing acute lethal toxicity. The third group
received furosemide (12.5 mg/kg i.p.) in 3.0 ml 0.9% NaCI
effects of furosemide and mannitol on the nephrotoxicity
and physiological disposition of COOP in the rat.
solution 30 mm prior to COOP (6 mg/kg i.v.); 4 hr after
Since data on BUN levels and renal histopathology did COOP administration, these animals were given 3.0 ml 0.9%
not always agree in our experiments (24), we evaluated
NaCI solution i.p. to reduce the possibility of excessive fluid
or electrolyte derangement. The last group received man
additional parameters of renal function in our studies.
Administration of other nephrotoxic compounds, such as nitol infusions, performed after animals were lightly anes
mercuric chloride and uranyl acetate, causes a decrease in thetized with pentobarbital (25 to 30 mg/kg i.p.) to facilitate
glomerularfiltration (9, 11). Therefore, we used [“ln]OTPA insertion of the 27-gauge x 0.5-inch needle of an infusion
blood clearance to estimate the effects of COOP alone or catheter set (EZ-Set Infusion Set; Deseret Pharmaceutical
with diuretics on glomerulan filtration rate. Because other Co., Sandy, Utah) into the lateral tail vein. The rats were
agents causing proximal tubular necrosis, such as paraquat
placed in restraining devices with their tails secured. After
or uranyl acetate, depress the organic acid secretion system the i.v. line was primed with 0.5 ml heparinized mannitol
solution, an infusion of 10% mannitol in 0.45% NaCI solu
in vivo (10, 11) and because the pars recta of the proximal
tion was begun at the rate of 0.10 mI/mm using a Harvard
tubule, the site of organic acid secretion (31), is apparently
most sensitive to COOP-induced damage in the rat (32), we Infusion Pump Model 975 (Harvard Apparatus Co. Inc.,
measured clearance of PAH as a further indication of Millis, Mass.). Following 25 mm of mannitol infusion, COOP
nephron function. In other experiments, we studied the (6 mg/kg) was injected as a bolus into the freely flowing i.v.
effects of furosemide and mannitol on the physiological
line. The infusion was continued for an additional 5 mm so
disposition of COOP by monitoring plasma, urine, kidney,
that rats received a total of 300 mg mannitol in 3.0 ml 0.45%
intestine, and spleen levels of platinum, the active center of NaCI solution.
COOP, at various time points. Through these measure
Collectionof PlasmaTissue, and Urine Samples. Four
series of experiments were performed to study BUN levels,
ments, we hoped to determine if diuretics were causing
major alterations in the disposition of COOP, a compound
kidney pathology, and platinum disposition. In Experiment
1, ratsin each treatmentgroup were housed in plastic
which is eliminated predominantly by the kidneys.
cages. Blood samples (150 to 200 @tI)
were obtained from
the
netroorbital
sinus
using
hepaninized
Natelson capillary
MATERiALS
AND METHODS
tubes 1 and 2 hr after COOP administration; these samples
Chemicals. COOPwas obtainedfrom the Drug Liaison were spun down in a Bninkmann Model 3200 Micro Centri
fuge (Brinkmann Instruments, Inc., Westbury, N. V.) to
and Distribution Section, Division of Cancer Treatment,
obtain plasma, which was later analyzed for platinum con
National Cancer Institute, NIH, Bethesda, Md. and diluted
to 1 mg/mI in 0.9% NaCI solution immediately prior to use. tent. Four days later, the animals were anesthetized with
ether, and blood was removed from the abdominal aorta
Furosemide, a gift of Hoescht-Roussel Pharmaceuticals,
Sommerville, N. J., was diluted so as to yield a dose of 12.5 using a heparinized syringe. After centnifugation, the
plasma was analyzed for platinum content and BUN. Fol
mg/kg in 3.0 ml sterile 0.9% NaCI solution. Mannitol,
obtained from Sigma Chemical Co., St. Louis, Mo., was lowing exsanguination of the rat, the left ventricle was
cannulated, and the tissues were slowly perfused with 0.9%
prepared as a 10% solution in 0.45% sterile NaCI solution.
NaCI solution. One kidney was fixed in 10% buffered for
[111InJOTPAwas purchased from Diagnostic Isotopes, Inc.,
Bloomfield, N. J. and diluted in 0.9% NaCI solution to an maim and sectioned at 5 to 6 @m,and the sections were
activity of 20 @Ci/mlat the time of injection. [3H]PAH was stained with hematoxylin-eosin and periodic acid-Schiff
The contralateral
kidneywas analyzedforplatinum
obtained at a specific activity of 1.26 Ci/mmol from New stains.
content.
England Nuclear, Boston, Mass. The compound was pre
Experiment 2 was performed in a similar fashion, except
pared as a 10 pCi/mI solution in 0.9% NaCI solution. Chloral
that blood samples were obtained prior to COOP adminis
hydrate was obtained from Sigma Chemical Co. and diluted
to 75 mg/kg in 0.9% NaCI solution. Pentobanbital was a tration and at 3 hr, and 3-, 4-, and 5-day time points
thereafter by retroorbital sinus bleeding. At 7 days, animals
product of Veterinary Laboratories Inc., Lanexa, Kans.
Drug Treatment. Male F344 rats weighing75 to 100 g were sacrificed, and terminal blood and kidney samples
were obtained as described above. The 0-, 3-, 4-, 5-, and 7were obtained from Microbiological Associates, Bethesda,
day plasma samples were analyzed for BUN; 3-hr samples
Md . or Charles River Breeding Laboratories, Wilmington,
were analyzed for platinum, and the kidneys were prepared
Mass. and fed a diet of standard lab chow (Wayne Lab
Blox, Allied Mills, Chicago, Ill.) and water ad !ibitum. They for platinum analysis and histopathology as described
were housed in groups of 5 to 6 animals in plastic cages above.
In Experiment 3, rats were housed in plastic metabolic
with corncob bedding (Experiments 1 and 2), stainless steel
wire bottom cages (renal function study and Experiment 4), cages (Maryland Plastics, Inc., New York, N. Y.). After an
or individually in plastic metabolic cages (Experiment 3). At adjustment period of 2 days, the rats were given the de
1270
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Diuretic Effects on CDDP Pharmacology and Renal Toxicity
scnibed drug treatments. Four, 8, and 12 mm after CDDP
administration, blood samples were obtained from the
retroorbital sinus. At 24 hr, the animals were sacrificed, and
blood and kidney samples were obtained as described
above. Plasma samples from each time point and one
kidney from each rat were analyzed for platinum content.
The contralateral kidney was prepared for histopathological
evaluation. Urine output and water intake volumes for the
24-hr period were determined, and an aliquot of urine
collected under mineral oil was removed for platinum anal
ysis.
In Experiment4, ratshoused in stainless
steel,wire
bottom cages were given COOP alone or with diuretics. At
2- and 15-mm and 1- and 2-hr time points, the animals were
sacrificed, and kidneys, spleens, and sections of proximal
duodenum were removed and rinsed in 150 mM NaCI
solution. Approximately 500 mg of each tissue was homog
enized in 10 ml ice-cold 150 mM NaCI solution. Two aliquots
were removed; one for determinations of platinum and a
second for measurement of protein by the method of Lowry
et al. (18). Results were then expressed in terms of ng of
platinum per mg protein.
@
@
where C is clearance in mI/mm, dose is total dpm adminis
tered, b1 is slope of slow terminal component, b2 is slope of
early rapid component, A is intercept of slow terminal
component, and B is intercept of early rapid component.
Clearance of PAH was estimated 3 days after drug treatment
using an i.v. dose of 15 @tCi
of [3H-PAH] per rat and a single
injection technique similar to that described above for
[“ln]DTPA(2).
Experiments 1, 2, and 3 were studied under light micros
copy and assigned a score which was a relative index of the
degree of tubular necrosis on a scale of 0 to 5 (0, no
necrosis; 5, extensive necrosis). The extent of tubular
epithelial
@
Platinum Assay, Determination of Radioactivity, and
BUN
Determination.
Platinum
was measured
by flameless
atomic absorption as described previously (22). “Inwas
counted in a Model 1195 Automatic ‘y
Counting System
(Amersham/Searle, Arlington Heights, III.) using a window
from 125 to 300 keV. Plasma samples of [3H]PAH were
counted in 10 ml of Biofluor aqueous scintillation fluid (New
England Nuclear, Boston, Mass.) using a Beckman LS-255
Liquid Scintillation Counter (Beckman Instruments, Inc.,
Fullerton, Calif.) and external standard quench calibration.
BUN was determined by colorimetry using the diacetylmo
noxime reagent (6).
Statistical Analysis. Results were analyzed first by single
factor analysis of variance; if a significant difference be
tween means was indicated, comparisons of means were
carried out using the Student-Newman-Keuls multiple range
test (35). The histopathological ratings were evaluated us
ing nonparametric statistics, first by the Kruskai-Wallis test
and then by the Mann-Whitney U test with error rate set on
Renal Function Experiments. To estimate glomerular a pen experiment basis (16). Differences were considered
filtration rates, we measured clearance of [“ln]DTPA(4, significant at the level ofp < 0.05.
28) using a single injection clearance technique (1, 2).
Three days after drug treatment, rats (housed in stainless
RESULTS
steel, wire-bottom cages) were anesthetized with chloral
Body Weight and 24-Hr Fluid Balance. Chart 1 shows
hydrate (300 mg/kg i.p.) and given injections of 10 pCi of
results from Experiment 2, where the nadir of weight loss in
[1111n]DTPA i.v. Blood samples (100 @l)
wene obtained at 5,
treated rats occurred on Day 5 (5 days after administration)
10, 15, 20, 40, 50, 60, and 70 mm by retroorbital puncture,
and at the final point, the rats were sacrificed. After plasma with body weights 88.8 ±3.4%, 83.2 ±1.9%, and 89.8 ±
4.5% (S.E.) of Day 0 weights in rats given COOP, COOP with
was obtained and counted as described below, a biphasic
furosemide, and COOP with mannitol, respectively. By Oay
plasma decay curve was plotted and clearance calculated
using the formula
7, all rats were gaining weight. No significant differences in
weight loss among treatment groups were noted, although
C —dose x b1b2
furosemide-treated rats consistently lost more weight. The
Ab2 Bb
Renal Histopathology.Stainedsectionsof kidneysfrom
@
toxicity described in an accompanying paper (23). These
kidneys were processed and examined in the same fashion,
and we report the results here for purposes of confirmation.
regeneration
was graded in a similar fashion. The
sections were evaluated on at least 2 different occasions; in
no case did the score vary more than one grade between
examinations. When variance occurred, the 2 scores were
averaged. The observer scoring the slides was unaware of
the code identifying the treatment group to which each
specimen belonged . Because histopathological
results
from this set of experiments
did not correlate
with renal
function studies, we also studied kidneys from rats given
identical treatments in a later experiment on bone marrow
120
0
110
z
0
U
0
>.
100
4
0
0
@90
I..
I
80
0
0
70
0
TIME POST TREATMENT
IDAYSI
Chart 1. Effect of COOP (6 mg/kg iv.) alone or with diuretics on body
weight. Data are from Experiment 2. Values are mean weights relative to Day
0 ±SE. , versus time after drug injection in days, with n = 6 for each group.
0, control; •,COOP; A, COOP with furosemide; •,COOP with mannitol.
There are no significant differences among treatment groups.
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1271
M. F. Pera, Jr. et a!.
inhibition of weight gain seen in control animals during
Days 3 to 5 may be due to the repeated bleeding from the
retroorbital sinus, a procedure which induces a degree of
stress (21). In a later experiment reported in the following
paper where no bleeding was performed, it was found that
recovery of body weight in mannitol-treated rats signifi
cantly outpaced that from other treatment groups during
this period (23).
Chart 2 from Experiment 3 indicates fluid output (as urine
volume) and fluid intake (as water consumption plus injec
tions or infusions) for the 24-hr period following drug
administration . Mannitol and furosemide groups excreted
significantly greater urine volumes (p < 0.05 by the multiple
range test) compared to COOP or 0.9% NaCI solution
treated control groups. The apparent positive fluid balances
seen are due to evaporation
100
a,
E
z
of urine and the fact that
insensible fluid loss is not measured. However, since the
net fluid balance appears similar for the groups, it is
unlikely that diuretic-treated animals were significantly de
hydrated due to increased urine output. Evaluation of fluid
balance over the short period immediately following COOP
administration proved difficult because of variable urine
output in animals not given diuretics. However, in other
studies, we were able to show that hematocnit values and
plasma chloride concentrations were within control limits
at the time of COOP administration.
Renal Toxicity: BUN Measurements.Serial BUN deter
minations from Experiments 1 and 2 are shown in Chart 3.
A dose of COOP (6 mg/kg i.v.) produced significant eleva
tion of BUN above control levels to a peak of 98.8 ±24.0
mg/dl on Day 5, followed by incomplete recovery toward
control levels on Day 7. Furosemide and mannitol groups
had BUN levels that were elevated, although not signifi
TIME
POST TREATMENT
IDAYSI
Chart 3. Effect of COOP (6 mg/kg iv.) alone or with diuretics on BUN
levels. Data are from Experiments 1 and 2. Values are mean BUN in mg/dl ±
SE. versustime in daysafter drug injection,with n = 6 except as noted in
parentheses. 0, control; C, COOPalone; A, COOPwith furosemide; U,
COOPwith mannitol. @,
valuessignificantly larger than other groups; p <
0.05.
cantly, above control; the BUN levels in these rats were
significantly
below those in rats given COOP alone (COOP
> furosemide,
mannitol,
or control
< 0.05 by the multiple
range test).
on Days 3, 4, 5, and 7; p
Renal Histopathology.Aftertreatmentwith COOP(6 mg/
kg i.V.) alone or with diuretics, a mild degree of early
degenerative changes was observed on Day 1. The lesions
were limited to epithelial cells of the straight proximal
E
‘U
tubules in the outer stripe of the outer zone of the medulla.
-I
0
>
£
0
CONTROL
CDDP
COOP
WITH
FUROSEMIDE
CDDP
WITH
MANNITOL
Chart 2. Effect of COOP (6 mg/kg iv.) alone or with diuretics on 24-hr
fluid balance. Data are from Experiment 3. Values for water intake and urine
output are mean ±S.E. with n = 6, except as noted in parentheses. @,
urine
outputs significantly greater than control or COOP alone; p < 0.05.
1272
The changes consisted of early pyknosis and cytoplasmic
vacuolation with focal loss of the brush border and intralu
minalhyaline
casts.Lesionson Day 3 were more advanced,
consisting of pyknosis, karyorrhexis, karyolysis, and cyto
plasmic eosinophilia with partial loss of the brush border.
Necrotic epithelial cells were sloughed into tubular lumina.
In addition to the straight proximal tubules, a few convo
luted tubular cells were affected to a lesser degree. The
necrotizing changes were more advanced by Day 4 and
even more marked by Day 7. Many necrotic epithelial cells
dotted with chromatin debris were contained in the lumina
of the straight proximal tubules (Figs. 1 to 4). Lesions
outside of the outer stripe were much less marked, but
necrotic tubular cells were observed in the proximal tubules
in medullary rays and especially in tubules adjacent to the
capsule at the top of medullary rays. By Day 10, the
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Diuretic Effects on CDDP Pharmacology and Renal Toxicity
@
necrotizing changes were much less marked, but lumina of
affected tubules were dilated. Tubular dilation first ob
served on Day 7 was much more marked by Day 10 and
involved tubules in the outer stripe and, to a lesser degree,
tubules in medullary rays. Distal tubules (even in the outer
stripe) and collecting tubules, as well as glomeruli were
largely unaffected except an occasional pyknotic nucleus.
Regenerative changes were first observed in occasional
outer stripe proximal tubules on Day 3; they were much
more obvious by Day 4 and continued to involve more
tubules by Days 4 through 10. The regenerative cells had
large nuclei with prominent chromatin granules, nucleoli,
and extensive basophilic cytoplasm (Figs. 2 to 4). Such
regenerating epithelial cells were often seen flattened be
tween tubular basement membranes and dying proximal
tubular epithelia. As the necrotic epithelia sloughed in the
lumina, the large regenerating cells took their places (Fig.
4). Some of the regenerative cells underwent karyorrhexis
and death; a few appeared to die during mitosis.
Chart 4 plots the course of the indices of necrosis from
individual rats, as well as the median indices, as a function
of time for the 3 treatment groups. Treatment with diuretics
did not have a noticeable effect on the site of necrotizing
changes within the kidney. There were no significant differ
ences in the degree of tubular necrosis on Days 1, 3, or 4.
Thereafter, particularly on Days 5 and 7, there was a trend
towards longer persistence of necrotizing processes in
furosemide-pretreated rats, and a tendency for mannitol
pretreated animals to display less damage compared to rats
given COOP alone (Figs. 2 to 4). Analysis of variance
showeda difference
inthe3 groupson Days5 and 7 (p<
0.05 by the Kruskal-Wallis test), but the data were insuffi
cient to allow a localization of the difference within groups
(per experiment, p > 0.05 by the Mann-Whitney U test). The
time course of appearance of regenerative lesions tended
to be accelerated in mannitol-treated rats and to lag some
what in rats given furosemide. As necrotizing lesions sub
sided on Days 7 and 10 in rats receiving COOP with
mannitol, there were also relatively fewer regenerating cells
compared to the other treatment groups. Because the
histopathological evaluation of kidneys from Experiments
1, 2, and 3 was in conflict with the observed protection
against BUN elevation, another set of kidneys from an
experiment in which rats were given identical drug treat
ments (23) was also evaluated histopathologically. The
above description and Chart 4 incorporate results from this
latter study.
GlomerularFiltrationand PAHClearance. BecauseBUN
0
U
S
0
K
a
S
TIME POST TREATMENTIDAYSI
Chart 4. Effect of diuretics on the degree of proximal tubular necrosis
after COOP(6 mg/kg iv.) Dataare from Experiments1, 2, and 3, and one
additional experiment. Values are indices of proximal tubular necrosis
versustime in daysafterdrug injectionon a scalefrom 0 = no necrosisto 5
=
extensive
necrosis.
Individual
ratings
for
well
as
median
ratings
COOP(6were given injections of 0.5 ml 0.9% NaCI solution or
glomerularfiltration
mg/kg iv.) alone or with diuretics. Three days later,
clearanceusing
rate was estimated
from [“ln]DTPA blood
a single injection technique (1 , 2).[1
clearanceTreatment
ln]OTPA
(ml/min/loogbodywt)Control n
±COOP
6
0.926
0.00629k'COOP
8bCOOP
with furosemide
6
5
0.0752±
0.529 ±0.11
with mannitol
5
a Values are mean ±S.E. determined
b Control > COOP plus furosemide
0.294 ±
3 days posttreatment.
> COOP plus mannitol
>
COOP;p < 0.05.
Table 2
clearanceRats
Effect of CDDPalone or with diuretics on PAH
COOP(6
were given injections of 0.5 ml 0.9%NaCIsolution or
PAHclearance
mg/kg i.v.) alone or with diuretics. Three days later,
radiolabeledcompound
was estimated from blood clearanceof the
using a single injection technique (2).PAH
clearanceTreatment
wt)Control
n
(ml/min/100 g body
0190a.bCOOP
5
2.59 ±
0•084bCOOP
5
0.530 ±
0.225@@COOP
with furosemide
with mannitol
5
5
1.59 ±
1.57 ±0.186@@
a Values are mean ±SE. determined
furosemide
CODP;p <0.05.
ance of [“ln]DTPA, was markedly
than that of untreated
controls
3 days after
as
Table 1
Effect
rateRats
of CDDPalone or with diuretics on glomerular filtration
with
depressed
rat
COOPwith furosemide;U, COOPwith mannitol.
measurements and histopathological results did not come
late, additional parameters of renal function were meas
ured. Glomerular filtration, estimated from plasma clear
COOP (6 mg/kg) (Table 1). Rats given furosemide or man
nitol with COOP had glomerular filtration rates which were
significantly higher than those in animals given COOP alone
but also significantly lower than those of untreated controls
(COOP < mannitol < furosemide < control; p < 0.05 by the
multiple range test). Clearance of PAH (Table 2) was also
significantly depressed by COOP. Again, diuretic treatment
significantly lessened the severity of the depression, but the
PAH clearance rate in diuretic-treated rats was still lower
each
are shown. Symbols for individual ratings: o•,COOP only; E@A,COOP with
furosemide; DU, COOP with mannitol (0, t@,and 0 and •,A, and U are
from 2 separate series of experiments). Median ratings: C, COOP only; A,
b Control
>
COOP
3 days after treatment.
or
COOP
with
mannitol
>
(COOP < furosemide
=
mannitol < control; p < 0.05 by the multiple range test).
Thus, diuresis as induced in these studies partially pro
tected renal function, as measured by these parameters.
Effect of Diuresison the Dispositionof Platinum.The
plasma decay of platinum following a dose of COOP (6 mg/
kg) is illustrated in Chart 5, a composite curve constructed
from
Experiments
1, 2, and 3. Tniphasic
decay
was ob
served, with half-lives of 8.8 and 49 mm and 1.7 days for a,
13,and y phases in animals given COOP only. Neither
APRIL 1979
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 1979 American Association for Cancer Research.
1273
M. F. Pera, Jr. et al.
z
‘U
0
a.
a,
E
a.
U)
4
0
0
z
4
z
S
4
TIME POST TREATMENT
TIME
POST
TREATMENT
(MIN)
ml.)
Chart 5. Effect of diuretics on plasma decay of platinum after COOP (6
mg/kg iv.) Composite curves constructed from Experiments 1, 2, and 3.
Valuesare mean plasmaplatinum concentrationsin pg/mI ±SE. versus
time in mm (lowerpanel) or hr (upperpanel)with n = 6 exceptas noted in
parentheses.Upperpanel, continuation of lower panel on a different time
scale with 180-mm points replotted. •,
COOP alone; A, COOPwith furosem
ide; U, COOPwith mannitol. @,
values significantly greater than COOP alone,
p < 0.05.
z
‘U
0
a.
E
a.
Cl)
diuretic
@
had marked effects
on the rate of plasma decay.
Significantly higher plasma platinum levels during the sec
ond phase of plasma decay were observed in furosemide
treated rats compared to rats receiving COOP only for 2and 3-hr time points (p < 0.05 by the multiple range test).
Mannitol treatment did not alter plasma platinum levels
significantly. The lower plasma platinum levels seen in this
group at 4, 8, and 12 mm may be the result of an expansion
of the central compartment by rapid infusion of hypertonic
solute.
Chart 6 shows results from studies of platinum levels at
early time points following administration of COOP in
spleen, small intestine, and kidney, all of which are suscep
tible to COOP toxicity. Significantly lower spleen platinum
levels were measured in both furosemide and mannitol
groups than in spleens from rats receiving COOP alone 2
mm after COOP (p < 0.05 by multiple range test). Thereaf
ten, no significant differences were seen in platinum content
among the 3 treatment groups in spleen or in small intestine
at any time points. In contrast, kidneys from mannitol
treated rats had significantly lower platinum levels than did
those of the other groups 2 mm after COOP administration.
At 15 mm post-COOP injection, there were no differences
among the groups. However, by 60 and 120 mm, kidneys
from funosemide-treated rats had significantly higher plati
num content than did kidneys from rats given either COOP
alone or COOP plus mannitol. This latter finding is consist
ent with more extensive studies at later time points (below).
On the basis of data on the amount of platinum associated
with acid-insoluble material, it appears that the slow rate of
loss of platinum from the above tissues was the result of
rapid binding to macromoleculam components and that the
diuretics did not affect this process. However, much addi
tional work is required to substantiate this conclusion.
Table 3 shows kidney platinum levels at 24 and 96 hr
1274
4
C,
0
z
4
z
TIME POST TREATMENT(MIN)
z
‘U
0
a.
E
a:
Cl)
C,
4
0
z
4
z
TIME
POST TREATMENT(MIN)
Chart 6. Effect of diuretics on spleen (a), small intestine (b), and kidney
(c) platinumcontent at earlytime pointsafterCOOP(6 mg/kg i.v.). Dataare
from Experiment 4. Values are mean tissue platinum concentrations in ng/
mg tissue protein ±SE. versus time in mm, with n = 3 except as noted in
parentheses. •,COOP alone; A, COOP with furosemide, U, COOP with
mannitol. @,
values significantly higher than those of all other treatment
groups.
values significantly lower than those of all other treatment
groups (p < 0.05 by multiple range test).
CANCER RESEARCHVOL. 39
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 1979 American Association for Cancer Research.
Diuretic Effects on CDDP Pharmaco!ogy and Renal Toxicity
post-drug administration . Kidneys from furosemide-treated
animals contained significantly more platinum at 24 hr than
did those from mannitol-treated rats (p < 0.05 by multiple
range test). At 96 hr, kidneys
from the funosemide
group
contained more platinum than did those from animals given
COOP on COOP plus mannitol (p < 0.05 by the multiple
range test). Mannitol-treated animals did not differ signifi
cantly from those given COOP alone in terms of kidney
platinum content 24 on 96 hr after drug administration.
In Chart 7, and the cumulative 24-hr urinary excretion of
platinum and the 24-hr urine platinum concentration are
plotted, respectively. Roughly 40% of injected platinum was
recovered in 0 to 24 hr urine samples. Both furosemide and
mannitol tended to decrease, although not significantly, the
amount of platinum excreted in urine over a 24-hr period.
The 2 diuretics did cause a significant drop in the concen
tration
ofplatinumin0-to24-hoururinesamples,shown in
Chart 7b (COOP < furosemide = mannitol; p < 0.05 by the
multiple mangetest). Thus diuresis did not cause a more
rapid plasma clearance, lower tissue platinum levels, or
increased urinary excretion of platinum but resulted in a
drop in urinary platinum concentration.
DISCUSSION
Cvitkovic et al. (7) showed that induction of mannitol
diunesis prevented the increase in BUN and serum creati
Rats were given injections of COOP(6 mg/kg iv.) alone or in nine caused by COOP administration to dogs. Ward et a!.
(34), on the basis of blood chemistry data and histopathol
combination
with diuretics and sacrificed 24 or 96 hr later. Kidneys
were removed,and platinum content was determined by flameless ogy, reported that when furosemide was administered to
atomic absorption spectrometry(22). Dataare from Experiments1 F344 rats prior to COOP, there was a decrease in the acute
and 3.
renal toxicity of the platinum compound, although toxicity
tissue24 (@.tg/gwet weight of
still occurred. Furthermore, the same investigators showed
that when COOP was administered in low weekly doses (3.0
hrnCOOP
TreatmentPlatinum
hrn96
mg/kg i.v.) for periods of 5 to 6 weeks, the renal failure that
Table 3
Effect of diuretics on kidney levels of platinum after 6 mg/kg iv.
COOPwithfuro
12.1 ±0.93c 5
6
885÷0598b
semide
COOPwith man
nitolg.64±1.10a
6.17 ±0.8l6@@6 6
6.27 ±1.38k'6 65,44±0.4l8@@
a Values are mean ±S.E.
b COOP with furosemide
> COOP with mannitol
or COOP; p <
0.05.
C
COOP
with
furosemide
>
COOP
A
with
mannitol;
p
<
0.05.
B
150
30 r
P
developed
in rats given COOP alone
(33). We shall restrict our comments
was not
to acute toxicity
after
a single, high, nonlethal dose because it is possible that the
pathogenesis of chronic renal failure induced by COOP is
different from that seen after acute intoxication. We chose
to study a high dose in light of current clinical trends
towards escalation of COOP dosage in combination with
diuretics
(3,13,25).
Our data on BUN levels confirm previous work. Two
additional parameters were studied to confirm that renal
function was protected in rats. Glomerular filtration rate
was shown to be depressed markedly 3 days after COOP
administration. Similar findings have been reported for
other nephrotoxic
or umanyl nitrate
a.
compounds,
such as mercuric chloride
(9, 11). Diuretic treatment partially pre
vented this decrease in filtration. Clearance of PAH was
also greatly diminished 3 days after COOP administration.
a,
z
CDDP WITH
0@
The
cells of the straight portion of the proximal tubule, the
FUROSEMIDE
z
predominant site of COOP toxicity in the rat (32), are
0
CDDP WITH
20 @.@ 100
Iresponsible for organic acid secretion (31), and PAH clear
MAN NITOL
4
ance
is depressed in vivo by other compounds that damage
I.
z
the proximal tubule (10, 11). Rats given diuretics with COOP
‘U
U
z
cleared PAH faster than did rats given COOP alone. This
0
U
faster clearance could be the result of improved function of
PAH-secreting cells on improved renal hemodynamics. We
a.
>-..
50
‘U
believe that it is important to point out that although these
z
4
-I
parameters of renal function were partially protected and
although BUN levels in diuretic-treated rats were not sign if
C.,
icantly higher than controls, both glomerular filtration and
PAH clearance were significantly lower in diuretic-treated
groups compared to untreated controls. Thus BUN level is
a reliable but relatively insensitive indicator of filtration
Chart 7. Effect of diuretics on cumulative urinary excretion (a) and con
function. When Dentino et a!. (8) studied the long-term
centration (b) of platinum in urine during a 0- to 24-hr collection period after
COOP(6 mg/kg iv.). Dataarefrom Experiment3. Valuesaremeanrecovery effect of COOP without diuretics on human renal function,
of platinum in urine, expressed as g.tgplatinum per 24 hr per 100 g body
they also concluded that moderate increases in BUN or
weight ±SE. (a) or mean concentration of platinum in 24-hr urine sample.
plasma creatinine levels may be accompanied by substan
@.tg/ml±S.E. (b) with n = 6 except as noted in parentheses. @,
values
EJ CDDP
U.
0
@
ultimately
observed in rats receiving furosemide with each course
E
a,
1001
significantly lower than COOP alone; p < 0.05.
tial decrements
in glomerular
filtration
nate. We emphasize
APRIL 1979
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1275
M. F. Pera, Jr. et a!.
that the protection of function by diuretics in our studies
was partial, not complete.
Despite protection of renal function, the diuretic-treated
rats still exhibited tubular necrosis. The trend was toward
longer persistence of histological damage in the fumosemide
group and faster recovery in rats given mannitol relative to
rats given COOP alone. Our results here disagree with those
of Ward et a!. (34), who reported histological protection
with furosemide on an acute basis. Theme are 2 possible
reasons for the discrepancy. First, Ward et a!. reported
histological data on rats given a slightly lower dose of
COOP (4.8 versus 6.0 mg/kg i.v. in the present work).
Second, Ward et a!. reported histological data only for
kidneys studied 3 days after drug exposure. Inspection of
Chart 4 shows that in our study on Day 3 the median
necrosis index for matsgiven fumosemide was in fact slightly
but insignificantly lower than that for rats given COOP
alone. Later, on Days 4, 5, 7, and 10, the median necrosis
index for the furosemide group exceeded those of the other
groups. In Chart 3, it may be seen that BUN levels also
peaked later in furosemide-treated mats. Larger studies
would be required to statistically verify the differences in
the degree of necrosis between the groups. Nonetheless,
the persistence of the COOP-induced lesions in fumosemide
treated rats 9bserved in 2 experiments was disturbing, as
was the tendency of this group to lose more weight.
Fumosemide is a potent diuretic agent with toxicities that
overlap those of COOP. This diuretic drug has been shown
to produce ototoxicity in humans (27), and it was recently
demonstrated that at high doses the compound can un
dergo a process of metabolic activation in animals to
reactive intermediates which bind covalently to macromol
ecules, resulting in proximal tubular necrosis (19). Unless
further study proves that themeis any advantage to the use
of furosemide, these data suggest that fumosemide should
be contraindicated for use in prevention of COOP-induced
nephmotoxicity. It would, therefore, seem prudent to use
other
compounds
for prevention
of COOP-induced
renal
failure
inhumans.
The contradiction between renal structure and renal func
tion measurements observed in these studies is not unpnec
edented. OiBonaetal. (9) compared chronically salt-loaded
matsgiven HgCI2 to rats given HgCl2 alone and found less
depression of glomerular filtration and less elevation of
BUN in the 0.9% NaCI solution-loaded animals, despite
equivalent degrees of proximal tubular necrosis. Further
more, Thiel et a!. (30) found that pretreatment with chronic
0.9% NaCI solution loading plus deoxycorticosterone ace
tate, acetazolamide, or furosemide protected mats from
mercuric chloride impairment of renal function but did not
alter the degree of necrosis in the straight portion of the
proximal tubule (30).
Our data on the kidney platinum
content
for the 3 treat
ment groups tend to correlate with the histopathological
pharmacological significance. A greater degree of persist
ent histological damage in the furosemide group was asso
ciated with a higher amount of platinum in the kidneys of
these animals. The protection of renal function by both
diuretics was accompanied by a drop in the urinary piati
num concentration, a phenomenon also observed in dogs
with mannitol (7).
Two recent reviews have discussed current concepts
concerning the pathogenesis of acute renal failure and the
possible association between tubular necrosis and depres
sion of filtration mate. It has been hypothesized that the
depression in filtration rate might be associated with back
leak of normally nonreabsombed substances through dam
aged epithelium, obstruction of tubules by debris, persist
ent vasoconstriction in the afferent arteriole secondary to
activation of the intmamenalmenin-angiotensis system, or a
direct effect on the glomerulus to decrease the glomerular
capillary ultrafiltration coefficient (17, 29). Backleak of
nonreabsorbed substances through damaged epithelium is
unlikely to account for our results, since the damage to the
tubule was similar in the diuretic-treated animals that ex
hibited improved filtration. However, any of the other path
ophysiological alterations might be triggered by high tubu
lamlevels of toxins such as COOP, and dilution
of the urine
could ameliorate the ensuing decrease in renal function.
Further studies are required to investigate these possibili
ties, but we mention them to illustrate that the diluting
effect of diuretics might reduce functional impairment of
the kidney without markedly altering the course of tubular
necrosis.
The concentration of platinum in plasma and intestine
was not altered by diuretic administration, nor did the
diuretics
increase the total urinary excretion
of platinum.
Cvitkovic et a!. (7) reported that mannitol diumesis did not
affect serum platinum levels in dogs, although a direct
comparison of dogs given COOP alone to mannitol-tmeated
dogs was not provided. Another report mentioned unpub
lished findings
of increased
blood levels of platinum
with
fumosemide in mats(34). Although we have noted a tendency
toward higher plasma platinum levels in fumosemide-treated
rats, particularly in their later phases of plasma decay, the
effect was not marked. Our studies on intestine and all but
the 2-mm time point on spleen showed no difference in
uptake of platinum into these tissues, consistent with the
plasma clearance data. No significant alterations in urinary
recovery of platinum were observed with furosemide or
mannitol, similar to previous reports with mannitol in dogs
and fumosemide in rats (7, 34). Thus, it seems unlikely that
mannitol
or furosemide
diuresis markedly alters the overall
cumulative disposition of platinum outside of the kidney. If
the level of platinum in plasma, spleen, intestine, and urine
reflects the disposition of the cytotoxic form(s) of COOP,
the preceding results suggest that combined administration
of diuretics with COOP should reduce renal toxicity without
results. Kidneys of mats given COOP alone or with mannitol
altering toxicity to host renewing cell populations
contained similar amounts of platinum and exhibited a
similar degree of necrosis. While the apparent more rapid
recovery from tubular necrosis in mannitol-treated rats
compared to matsgiven COOP alone correlates with sign ifi
cantly lower 2-mm kidney platinum levels, further investi
gations are required to substantiate and determine its
testinal mucosa and bone marrow), and without changing
1276
antitumor
activity.
The accompanying
publication
(gastmoin
will ad
dressthesemattersdirectly.
REFERENCES
1. Blaufox, M. 0., and Cohen, A. Single injection clearance of iothalamate
CANCER
RESEARCH
VOL. 39
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 1979 American Association for Cancer Research.
Diuretic Effects on CDDP Pharmacology and Renal Toxicity
I in the rat. Am. J. Physiol., 218: 542-544, 1970.
2. Blaufox,M. 0., Guttmann,R. 0., and Merrill,J. P. Measurementof renal
function in the rat with single injection clearances. Am. J. Physiol. , 212:
629-632,1967.
3. Chary, K. K., Higby, 0. J., Henderson, E. S., and Swinerton, K. 0. Phase
I study of high-dose cis-dichlorodiammineplatinum(ll)
with forced di
uresis. CancerTreat. Rep., 61: 367-370, 1977.
4. Chervu, L. R., Lee, H. B., Goyal, 0., and Blaufox, M. 0. Use of―Tc-Cu
diammine platinum (therapeutic efficacy in previously treated wide
spread and recurrent testicular tumors). Proc. Am. Assoc. Cancer Res.,
17:243,1976.
21. Migdalof, B. H. Methods for obtaining drug time course data from
individual small laboratory animals: serial microblood sampling and
assay. Drug Metab. Rev., 5: 295-310, 1976.
22. Pera,M. F., andHarder,H.C. Analysisfor platinumin biologicalmaterial
by flameless atomic absorption spectrometry. Clin. Chem., 23: 1245-
1249,1977.
DTPAcomplexasa renalfunction agent.J. NucI.Med.,18:62-66,1977.
5. Connors,T. A. Antitumoreffectsof platinumcompoundsin experimental 23. Pera,M. F., and Harder,H.C. Effectsof mannitolor furosemidediuresis
animals. In T. A. Connors and J. J. Roberts (eds.), Platinum Coordination
on cis-dichlorodiammineplatinum(ll)
Complexes in Cancer Chemotherapy,pp. 113-123. Berlin: Springer
Verlag,1974.
host-renewingcell populations in rats. Cancer Res., 39: 0000-0000.
6. Crocker, C. L. Rapid determination of urea nitrogen in serum or plasma
without deproteinization.Am.J. Med.Technol.,33: 361-365,1967.
7. Cvitkovic, E., Spaulding, J., Bethune, V., Martin, J., and Whitmore, W.
F. Improvement of Cis-dichlorodiammineplatinum (NCS 119875): thera
peutic index in an animal model. Cancer, 39: 1357-1361 , 1977.
8. Oentino, M., Luft, F. C., Vum, M. N., Williams, S. C., and Einhorn, L. H.
Long term effect of cis-diamminedichloride platinum (COOP) on renal
function and structure in man. Cancer, 41: 1274-1281 , 1978.
9. OiBona, G. F., McDonald, M. 0., Flamenbaum, W., Oammin, G. J., and
Oken, 0. E. Maintenanceof renal function in salt loaded rats despite
severe tubular necrosis induced by [email protected], 8: 205-220, 1971.
10. Ecker, J. L., Hook, J. B., and Gibson, J. E. Nephrotoxicity of paraquat in
mice. Toxicol. AppI. Pharmacol., 34: 178—186,
1975.
11. Foulkes,E. C. Glomerularfiltration and renal plasmaflow in uranium
poisoned rabbits. Toxicol. AppI. Pharmacol., 20: 380-385, 1971.
12. Gonzalez-Vitale,J. C., Hayes,0. M., Cvitokovic,E., andStemberg,S. S.
The renal pathology in clinical trials of cis-platinum (II) diammine
dichloride. Cancer, 39: 1362-1371, 1977.
13. Hayes, 0. M., Cvitkovic, E., Golbey, R. B., Schemer, E., Helson, L., and
Krakoff, I. H. Highdosecis-platinumdiamminedichloride: amelioration
of renal toxicity by mannitol diuresis. Cancer, 39: 1372-1381 . 1977.
14. Kociba, R. J., and Sleight, S. 0. Acute toxicologic and pathologic effects
of cis.diamminedichloroplatinum(NSC-1119875)in the malerat. Cancer
Chemother. Rep., 55 Part 1: 1-8, 1971.
15. Kociba, R. J., Sleight, S. 0., and Rosenberg, B. Inhibition of dunning
ascitic leukemia and Walker 256 carcinosarcoma with cis-diamminedi
chloroplatinum (NCS-119875). Cancer Chemother. Rep., 54: 325-328,
1970.
16. Lehmann,E. L. Nonparametrics:statistical methods basedon ranks.
San Francisco: Holden-Oay. Inc., 1975.
17. Levinsky, N. G. Pathophysiology of acute renal failure. N. EngI. J. Med.,
296: 1453-1458, 1977.
18. Lowry, 0. H., Rosebrough, N. J., Farr, A. L., and Randall, R. J. Protein
measurementwith the Folin phenol reagent.J. Biol. Chem.,193:265275,1951.
19. McMurty, R. J., and Mitchell, J. R. Renal and hepatic necrosis after
metabolic activation of 2-substituted furans and thiophenes, including
furosemide and cephaloridine. Toxicol. AppI. Pharmacol., 42: 285-300,
1977.
20. Merrin, C. A new method to prevent toxicity with high doses of cis
antitumor activity and toxicity to
1979.
24. Pera,M. F., and Harder,H. C. Effectsof furosemide(FM)-and mannitol
(MN)-induceddiuresison the nephrotoxicityand physiologicaldisposi
tion of cis-dichlorodiammineplatinum (COOP) in rats. Proc. Am. Assoc.
Cancer Res., 19: 100, 1978.
25. Rozencweiz, M., Von Hoff, 0. 0., Slavik, M., and Muggia, F. M. Cis
diamminedichloroplatinum
(II): A new anticancer drug. Ann. Intern.
Med.,86: 803—812,
1977.
26. Schaeppi, U., Heyman, I. A., Fleischman, R. W., Rosenkrantz, H.,
Ilievski, V., Phelan, R., Cooney, 0. A., and Oavis, R. 0. Cis-dichlorodiam
mineplatinum(ll) (NCS-119875): Preclinical toxicologic evaluation of in
travenous injection in dogs, monkeys and mice. Toxicol. AppI. Pharma
col.,35: 230-241, 1973.
27. Schwartz, G. H., David, 0. S., Riggio, R. R., Stenzel, K. H., and Rubin,
A. L. Ototoxicity induced by furosemide.N. EngI. J. Med., 282: 14131414, 1970.
28. Sziklas,J.J., Hosain,R., Reba,A. C., andWagner,H.N.,Jr. Comparison
of ‘‘Vb-OTPA,
llamln@0TPA,‘4C-inulinand endogenous creatinine to
estimate glomerular filtration. J. NucI. Blol. Med., 15: 122-125, 1971.
29. Stein, J. H., Lifschitz, M. 0., and Barnes, L. 0. Current concepts on the
pathophysiology of acute renal failure. Am. J. Physiol., 234: F171-F181,
1978.
30. Thiel, F., Brunner, F., Wunderlich, P., Huguenin, M., Bienko, B.,
Torhorst, J., Peters-Haefeli, L., Kirchertz, E. J., and Peters, B. Protection
of rat kidneysagainstHgCI,-inducedacute renal failure by induction of
high urine flow without renin suppression. Kidney Int., 10: S191-S200,
1976.
31. Tune, B. M., Burg, M. B., and Patlak, C. S. Characteristics of p-amino
hippurate transport in proximal renal tubules. Am. J. Physiol. 217: 19571963,1969.
32. Ward,J. M., and Fauvie,K. A. Thenephrotoxiceffectsof cis-diammine
dichloroplatinum(II) (NCS-119785) in male F344 rats. Toxicol. AppI.
Pharmacol., 38: 535-547, 1976.
33. Ward, J. M., Grabin, M. E., Berlin, E., and Voung, 0. M. Prevention of
renal failure in rats receiving cis-diamminedichloroplatinum(Il)
by ad
ministration of furosemide. Cancer Res., 37: 1238-1240, 1977.
34. Ward, J. M., Grabin, M. E., Leroy, A. F., and Voung, 0. M. Modification
of the renal toxicity of cis-dichlorodiammineplatinum(lI) with furosemide
in male F344 rats. Cancer Treat. Rep., 61: 375-379, 1977.
35. Zar, J. H. Biostatistical Analysis. Englewood Cliffs, N. J.: Prentice-Hall,
Inc., 1974.
Figs. 1 to 4 were taken from the outer stripe of the outer zone of the renal medulla 7 days after the indicated treatments. The slides were stained with
periodic acid-Schiff and photographed at x 400.
Fig. 1. Untreated control. Histological grade of necrosis = 0. No lesions are observed.
Fig. 2. COOP.Histologicalgradeof necrosis= 3. Straightarrows, moderatedegreeof necrosisindicatedby focal dissolutionof proximaltubular cells.
Curved arrow, regenerated cells have large nuclei.
Fig. 3. COOP with furosemide. Histological grade of necrosis = 4. Marked necrosis of straight proximal tubular cells is revealed by pyknosis, cytoplasmic
disintegration and sloughing into lumina. Enlarged nuclei (curved arrows) indicate regenerative efforts. The proximal convoluted tubules, with dark-staining
brush border (top) and distal tubules (thick arrow) are essentially normal.
Fig. 4. COOP with mannitol. Histological grade of necrosis = 1. Mild degree of necrotizing changes is indicated by pyknotic cells and nuclear and
cytoplasmic debris, which are forced into lumina by regenerating cells (curved arrows).
APRIL
1979
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 1979 American Association for Cancer Research.
1277
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CANCER RESEARCH VOL. 39
Downloaded from cancerres.aacrjournals.org on June 18, 2017. © 1979 American Association for Cancer Research.
Effects of Mannitol or Furosemide Diuresis on the
Nephrotoxicity and Physiological Disposition of cis
-Dichlorodiammineplatinum-(II) in Rats
Martin F. Pera, Jr., Bernard C. Zook and Harold C. Harder
Cancer Res 1979;39:1269-1278.
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