[CANCER RESEARCH 47, 1239-1243, March 1, 1987]

Role of Phospholipase in the Genesis of Doxorubicin-induced Cardiomyopathy in

Rats
Yuuichi Ogawa, Taizo Kondo, Satoru Sugiyama, Kouichi Ogawa, Tatsuo Satake, and Takayuki Ozawa1
Departments of Internal Medicine [Y. O., T. K., K. O., T. S.], and BiomédicalChemistry (S. S., T. OJ, Faculty of Medicine, University ofNagoya, Nagoya 466, Japan

ABSTRACT in the development of cellular dysfunction. In this study, we

investigate the mechanism of doxorubicin-induced cardiomy
The role of phospholipaseon the mechanismof doxorubicin-induced
opathy, especially in relation to phospholipase activity.
cardiomyopathywas investigatedin the heart mitochondriaof Wistar
rats.
In the in vivo study, rats were dividedinto 3 groups:/, the control MATERIALS AND METHODS
group,untreated;2, the doxorubicin1-daygroup,in whichdoxorubicin
In VivoStudy
(4 mg/kg)was injecteds.c. once;and3, the doxorubicin4-daygroup,in
whichdoxorubicin(4 mg/kg)was injectedonce a day for 4 consecutive Experiments were carried out on male Wistar rats weighing 200 ±
days. In each group,the level of lipid peroxidesand the phospholipase 30 (SD) g.
activity, the phospholipidcontent, and the enzymic activities in the Measurement of Concentrations of Doxorubicin in Heart and Plasma.
respiratorychainwere measured.The doxorubicin4-day groupshowed We determined the doxorubicin concentration in the heart and plasma
significantincreasesof lipid peroxidelevel and phospholipaseactivity by use of high performance liquid chromatography with fluorescence
and an inhibitionof mitochondria!respiratoryfunctioncomparedwith detector (9, 10). To measure the heart doxorubicin concentration, 5
the controlgroup,whilethe doxorubicin1-daygroupshowedno signifi rats were given injections of doxorubicin (4 mg/kg); doxorubicin-HCl;
cantdifference. Kyowa Hakko Kogyo Co., Ltd., Tokyo, Japan) once s.c. and 5 other
In the in vitrostudy,Experiment1, intactratheartmitochondriawere rats were given injections of doxorubicin (4 mg/kg) s.c. once a day for
incubatedwith 0.1 unit of phospholipaseA2. After the incubation,the 4 consecutive days. Twelve h after the last injection of doxorubicin,
enzymicactivitiesof the respiratorychain were disturbedin the same each rat was killed and the heart was removed. The heart was homog
manneras in the in vivoexperiment.In Experiment2, rat heartmito enized and extracted with a mixture of butyl alcohol and toluene. The
chondriawereincubatedwithascorbateandferroussulfate.The experi extract was evaporated in a vacuum, and the residue was dissolved with
ment demonstratedthe elevation of phospholipaseactivity associated a mixture of phosphate buffer and methanol. An aliquot of this solution
withlipidperoxidation. was injected onto a reverse-phase Ol >S' column and eluted with a
These results suggested that the enhancedphospholipaseactivity mixture of formic acid and methanol. The fluorescence signal was
causedby lipid peroxidationis responsiblefor the mechanismof doxo monitored at excitation wavelength 470 nm and emission wavelength
rubicin-induced cardiomyopathy. 585 nm.
To measure the plasma doxorubicin concentration, doxorubicin (4
INTRODUCTION mg/kg) was injected s.c. Blood samples were withdrawn into heparin-
ized syringes at 0.5,1,3,10, and 24 h after doxonibicin administration.
In 1967, Di Marco et al. (1) developed a new antitumor drug, The plasma samples adjusted to about pH 3 by l M 11,!'<),,. were
doxorubicin, which has been widely used because of its effec injected directly onto the column and eluted stepwise with phosphate
tiveness on many kinds of human neoplasm. Unfortunately, buffer and a mixture of the same buffer and acetonitrile. The chromat-
the dose-dependent and lethal cardiotoxic effect induced by ograms were monitored at excitation wavelength 470 nm and emission
doxorubicin has limited its clinical use. Minow et al. (2) dem wavelength 585 nm.
onstrated that cardiomyopathy was seen in up to one-third of Preparation of Rat Heart Mitochondria. Rats were divided into three
the patients receiving more than 600 mg/m2 of the drug. Many groups: the control group, untreated; the doxorubicin 1-day group,
investigators have proposed that the lipid peroxidation of car doxorubicin (4 mg/kg) was s.c. injected once; and the doxorubicin 4-
diac membrane played an important role in generating the day group, doxorubicin (4 mg/kg) was injected once a day for 4
cardiac dysfunction caused by doxorubicin (3-5). Doroshow (6) consecutive days. Rats were killed 12 h after the last doxorubicin
has established that the proximal portion of the NADH dehy- injection, and the hearts were removed. The cardiac mitochondria
fraction was prepared by the method of Hatefi et al. (11) and finally
drogenase complex [NADH: (acceptor) oxidoreductase, EC suspended in 0.25 Msucrose/10 mM Tris-HCl (pH 7.8) buffer (5-6 mg
1.6.99.3] was the site of anthracycline reduction in mitochon protein/ml). Due to the large amount of protein required for measure
dria and that the Superoxide formation was in a dose-dependent ment of lipid peroxide level, phospholipase activity, electron-transport
manner. As biomembrane consists of phospholipids and pro activity, and phospholipid content, segments from three hearts were
tein, it is deduced that the peroxidation of the fatty acids of the combined for preparation of the mitochondria fraction; that is, each
membrane phospholipids deteriorates the cellular dysfunction. group consisted of 24 rats, and 8 samples were obtained from each
Indeed, Dobretsov et al. (7) reported that the lipid peroxidation group.
increased the phospholipid bilayer rigidity and modified the Measurement of Mitochondria! Lipid Peroxides. Mitochondrial lipid
peroxides were determined by the method of Ohkawa et al. (12). The
physical state of the membrane resulting in the loss of its
outline of the procedure used is as follows: 0.2 ml mitochondria
function, whereas Iwata et al. (8) reported that when using rats suspension was mixed with 0.2 ml of 8.1% sodium dodecyl sulfate, 1.5
breathing pure oxygen, activation of phospholipase induced by ml of 20% acetic acid solution of pH 3.5, and 1.5 ml of 0.8% aqueous
lipid peroxidation is responsible for the genesis of oxygen- solution of thiobarbituric acid. The mixture was made up to 4.0 ml
induced lung injury. Thus, it is suggested that not only lipid with distilled water and heated at 95*C for 60 min. After cooling with
peroxidation but also the activtion of phospholipase is involved tap water, 1.0 ml of distilled water and 5.0 ml of a mixture of «-butyl
alcohol and pyridine (15/1, v/v) were added, then the mixture was
Received 6/26/86; revised 11/6/86; accepted 12/4/86. shaken vigorously. After centrifugation at 3000 rpm for 15 min, the
The costs of publication of this article were defrayed in part by the payment
of page charges. This article must therefore be hereby marked advertisement in absorbance of the organic layer (upper layer) was measured at 532 nm.
accordance with 18 U.S.C. Section 1734 solely to indicate this fact. As an external standard, tetraethoxypropane was used, and lipid per-
1To whom requests for reprints should be addressed, at Department of
BiomédicalChemistry, Faculty of Medicine, University of Nagoya Tsuruma, 1 The abbreviations used are: ODS, octadecyl silane; ADAM, 9-anthryldiazo-
Showa-ku, Nagoya 466, Japan. methane.
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 PHOSPHOLIPASE AND DOXORUB1CIN CARDIOMYOPATHY

oxide level was expressed in terms of nanomoles of malondialdehyde. mitochondria by the method of Bligh and Dyer (13) and then dried
Phospholipase Activity. Phospholipase activity of heart mitochondria under a stream of N and dissolved in chloroform. The phospholipid
was measured by the amount of fatty acids released from the substrate content in cardiac mitochondria was determined by the phosphorus
in the reaction mixture. One ml of mitochondria suspension was content of the extract according to the method of Chen et al. (18).
incubated 37*C with 40 nmol of L-a-phosphatidylcholine dilinolenoyl
(Funakoshi Pharmaceutical, Tokyo, Japan) as the substrate. After In Vitro Study
¡m ub.itum for 30, 60, and 90 min, linolenic acid was extracted by the
method of Bligh and Dyer (13). Heptadecanoic acid (12.5 nmol: Gas- Cardiac mitochondrial suspensions (5-6 mg protein/ml) were pre
kuro Kogyo, Tokyo, Japan) was used as an internal standard. The pared from intact rats and used in the following experiments.
extracted fatty acids were measured by the method of Shimomura et al. Experiment 1, Effect of Phospholipase \: on Mitochondrial Function.
Using cardiac mitochondrial suspension, changes in the electron-trans
(14). They were mixed with 0.2 ml of ADAM (Funakoshi Pharmaceu
tical, Tokyo, Japan) solution (0.5 g/liter methanol) for derivation. Theport activity induced by 0.1 unit of phospholipase A2 (\tija naja venom;
Sigma) were determined after 10 min of incubation with phospholipase
mixture was incubated for 3 h in the dark at room temperature and A2 at 30'C. The activities of the electron-transport chain divided into
analyzed by high performance liquid chromatography. Ten /il of ADAM
three segments (the NADH-cytochrome c reductase, the succinate-
derivatives of fatty acid solution were injected into a Shimadzu ODS
guard column (0.21 x 5 cm), (Shimadzu, Ltd., Tokyo, Japan) plus cytochrome c reductase, the cytochrome c oxidase) were determined by
Zorbax ODS analytical columns (0.46 x 15 cm plus 0.46 x 25 cm) at the same methods used in the /// vivo study.
60*C (back pressure: 80 kg/cm2; flow rate: 1 ml/min; solvent: methanol/ Experiment 2, Effect of Lipid Peroxidation on Phospholipase Activity.
Rat heart mitochondria were incubated at 37°Cfor 60 min. Incubations
water (94.7/ 5.3, v/v). ADAM derivatives of fatty acids eluted from the
column were detected by fluorescene spectromonitor (RF-500 LCA; were performed in 0.15 M KCI/20 mM Tris-HCl (pH 7.4) buffer
Shimadzu) connected to a computerized recorder (Chromatopac C- containing 0.1 mM ascorbate and 0.04 mM FeSO4. Control samples
contained neither ascorbate nor FeSO4. After the incubation, the phos
Rl A; Shimadzu).
Estimation of Mitochondria! Function. The activity of the electron- pholipase activity and the lipid peroxide content were measured by the
transport chain represents the mitochondrial function. The frozen- same methods used in the in vivo study.
thawed myocardial mitochondria used for the estimation of the elec Proteins were determined by the biuret reaction (19) using bovine
tron-transport chain was divided into three segments: the NADH —»serum albumin as a standard. All results in this paper are presented as
a mean ±SD. Analysis of variance with Dunnett's test was used for
coenzyme Q —» cytochrome <r,the succinate -* coenzyme Q —» cyto-
chrome <-,and the cytochrome c —»
cytochrome a, a, —»
O2. The enzymic statistical analyses of the data and was considered significant when P
activities of the NADH-cytochrome c reductase (NADH:ubiquinone was less than 0.05.
oxidoreductase, EC 1.6.99.3 and ubiquinol: ferricytochrome c oxido-
reductase, EC 1.10.2.2), the succinate-cytochrome c reductase (succi- RESULTS
nate:ubiquinone oxidoreductase, EC 1.3.99.1 and ubiquinol: ferricyto-
chrome c oxidoreductase, EC 1.10.2.2) and the cytochrome c oxidase In Vivo Study
(ferrocytochrome c:oxygen oxidoreductase, EC 1.9.3.1) were measured.
NADH-Cytochrome c Reductase. The specific activity of NADH- Concentrations of Doxorubicin in Heart. The concentrations
cytochrome c reductase was determined by a modified method of Hatefi of doxorubicin in each group are shown in Table 1. Doxorubicin
and Rieske (15). The reaction mixture consisted of 0.06 ml K..II!'().,- concentration in the doxorubicin 4-day group was elevated
HCI buffer ( 1.0 M, pH 8.0), 0.1 mi of NaN3 (0.1 M), 0.06 ml of EDTA remarkably compared with the doxorubicin 1-day group.
(1 mM), 5 ni of 1% deoxycholic acid (pH 8.0), 0.18 ml of 1% ferricy Concentrations of Doxorubicin in Plasma. Plasma concentra
tochrome c (Sigma Chemical Co., St. Louis, MO), and 2.6 ml of tion of doxorubicin is shown in Table 2. Doxorubicin concen
distilled water. The reaction was initiated by adding 10 /il of mitochon tration reached its peak at 30 min after the injection and
dria suspension and 75 /il of 10 mM NADH. After 15 s incubation at decreased time dependently. Negligible concentration was ob
30*C, the reaction rate was followed for 1 min by recording the increase
in absorbance of cytochrome c at 550 nm. The activity of NADH- served 24 h after the injection.
cytochrome c reductase was deduced from the increasing rate of the
Levels of Lipid Peroxides. The level of lipid peroxides in
absorbance. heart mitochondria in each group is shown in Fig. 1. In the
Succinate-Cytochrome c Reductase. The specific activity of succinate- control group, it was 3.86 ±0.75 nmol malondialdehyde/mg
cytochrome c reductase was determined by the method of Tisdale (16). protein. In the doxorubicin 4-day group, there was a significant
The reaction mixture consisted of 0.3 ml K2HPO4-HCI (100 mM, pH increase in the level of lipid peroxides (6.78 ±1.10) compared
7.4), 0.03 ml of NaNj (0.01 M), 0.06 ml of EDTA (0.01 M), 0.15 ml of
10% bovine serum albumin, 0.3 ml of potassium succinate (0.1 M), 0.3 Table 1 Concentrations of doxorubicin in tHe Heart
ml of 1% ferricytochrome c, and 1.86 ml of distilled water. The reaction Concentrations of doxorubicin in the heart were determined as described in
"Materials and Methods." Number of experiments in each group was five.
was initiated by adding 10 /¿Iof mitochondria suspension. In the same
way as with NADH-cytochrome c reductase, after 15s incubation at Concentration dig of
30'C, the reaction rate was followed for 1 min by recording the increase compound/g tissue, wet wt)
in absorbance of cytochrome c at 550 nm. The activity of succinate- Doxorubicin, 1 day 1.55 ±0.27*
Doxorubicin, 4 days 6.28 ±1.30*
cytochrome c reductase was deduced from the increasing rate of absor
°Mean ±SD.
bance. * Statistically significant, at P < 0.01, compared with doxorubicin 1-day group.
Cytochrome c Oxidase. The specific activity of cytochrome c oxidase
was determined by a modified method of Wharton and Tzagoloff(17).
To prepare ferrocytochrome c, 1% ferricytochrome c was reduced Table 2 Concentrations of doxorubicin in the plasma
completely by dithionite, and excess dithionite was removed by passing Concentrations of doxorubicin in the plasma were determined as described in
"Materials and Methods." Number of experiments for each time was four.
the solution through a column of Sephadex G-25 (fine), k 111'O4 l K I
buffer (2.67 ml; 50 mM, pH 7.0) and 30 »il of 10% Triton X-100 were Time after
injection
(h)0.5 Oig/ml)0.1
added to 100 ¿ilof ferrocytochrome c solution. Immediately after the 79 ±0.028°
addition of 0.1 ml mitochondria suspension, the reaction rate was
1 0.077 ±0.035
followed for 1 min by recording the decrease in absorbance at 550 nm. 3 0.039 ±0.006
The specific activity of cytochrome c oxidase was deduced from the 10 0.006 ±0.01 1
decrease of absorbance. 24Concentration Not detected
Measurement of Phospholipids. Lipids were extracted from cardiac ' Mean ±SD.

1240
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 PHOSPHOLIPASE AND DOXORUBICIN CARDIOMYOPATHY
~8-1 Table 3 Electron-transport activities in each group
I'-ì6lì:«i The enzymic acliviiies of Ihe eleclron-lransport chain were determined as
described in "Materials and Methods." Number of experimente in each group
was eighl.
protein/min)Control (nmol/mg

-,•5l
3S cytochrome cytochrome
—11'2o c reducÃ-ase c reducÃ-ase c oxidase
(pH
30'C)426
8.0, (pH30'C)377
7.4, (pH
30'C)2494
7.0,
56"507
± ±56 ±216
QTT1_Lcontro! Doxorubicin, 1 day ±121 406 ±49 2102 ±434
doxorubicin
doxorubicinone 205 ±29*Succinate-
Doxorubicin, 4 daysNADH- 1624±
358 ±51Cylochrome 125*
day(8) day 4 ' Mean ±SD.
(8) (8) * Slalislically signifìcanl,al P < 0.01, compared wiih conlrol.
Fig. 1. Hearl mitochondria! lipid peroxides coment in the conlrol group, Ihe
doxorubicin 1-day group, and the doxorubicin 4-day group. There was a significant
difference between the conlrol group and the doxorubicin 4-day group (/>< 0.01).
Mean ±SD (bars) of lipid peroxide content. Numbers in parentheses, number of
samples.

1000 8^20-||liII
900-
800
700-
15-en_L_LTn
600
500-
ncontroln
400- doxorubicin
doxorubicinone
If 300-
day(8)
(8)
day 4
(8)
¡I 200- Fig. 3. Heart mitochondria) phospholipid content in Ihe conlrol group, Ihe
doxorubicin 1-day group, and Ihe doxorubicin 4-day group. There was a significanl

I! 100-

0
30 60 90
decrease in the doxorubicin 4-day group compared with the control group (/' <
0.05). Mean ±SD (hur.\¡
of samples.
of phospholipid contenÃ-.Numbers in parentheses, number

Time(ran)
Table 4 Electron-transport activities with phospholipase I.-pretreatment
Fig. 2. Phospholipase activity measured from the released amount of linolenic
acid in each group. There was a significant increase in the doxorubicin 4-day Enzymic activities of the electron-lransport chain incubaled wilh 0.1 unit of
group compared wilh the control group (/' < 0.01). O, control group; •,doxorub phospholipase A¡were determined as described in "Materials and Methods."
icin 1-day group; •doxorubicin 4-day group. Number of experiments in each group was eighl.
protein/min)Control (nmol/mg
with the control group (P < 0.01), although the level of lipid
peroxides in the doxorubicin 1-day group was not significantly cylochrome cytochrome
increased compared with that of the control group. c reductase c reductase c oxidase
(pH30'C)413
8.0, (pH30'C)385
7.4, (pH
30'C)2404
7.0,
Phospholipase Activity. Fig. 2 illustrates the phospholipase ±62"273 ±50 ±203
activity in heart mitochondria, which was measured by the Phospholipase \ • ±70*Succinate- 358 ±56Cytochrome 1439 ±383*
amount of linolenic acid released from dilinolenoyl phosphati- pretreatmentNADH-
dylcholine. There was a linear relationship between the release " Mean ±SD.
* Statistical!) significant, al P< 0.01, compared with control.
of linolenic acid and the incubation time in each group. The
releasing rate of linolenic acid as phospholipase activity de
duced from 60-min values was 311 ±40 (ng/mg protein/h) in was a significant decrease in the doxorubicin 4-day group when
the control group. In the doxorubicin 4-day group, marked compared with the control group.
elevation in phospholipase activity was observed with values of In Vitro Study
521 ± 160 (P < 0.01), although there were no significant
changes in the doxorubicin 1-day group. Experiment 1. The activities of three segments in the mito
Mitochondria! Function. The activities of the electron-trans chondria! electron-transport chain with phospholipase A2 pre
treatment are shown in Table 4. The activities of NADH-
port chain in heart mitochondria in each group are shown in
Table 3. There was no significant difference between the activity cytochrome c reductase and cytochrome c oxidase were de
of NADH-cytochrome c reductase in the control group and that creased significantly compared with the control group without
of the doxorubicin 1-day group, but there was significant de phospholipase A2pretreatment. The activity of succinate-cyto
crease in the activity of the doxorubicin 4-day group. There was chrome c reductase showed a tendency to decrease, but the
no significant difference in the activity of succinate-cytochrome decrease was not significant.
c reductase among the three groups, although it showed a Experiment 2. The level of lipid peroxides and the activity of
tendency to decrease in the doxorubicin 4-day group. The phospholipase in mitochondria incubated with ascorbate and
FeSO4for 60 min at 37°C are shown in Fig. 4. After incubation,
doxorubicin 4-day group showed a significant decrease in the
activity of cytochrome c oxidase compared with the control the phospholipase activity and the level of lipid peroxides were
group, whereas there was no significant decrease in that of the elevated significantly compared with the control.
doxorubicin 1-day group.
Phospholipid Content. Phospholipid content in cardiac mi DISCUSSION
tochondria is shown in Fig. 3. Although the phospholipid Cardiotoxicity caused by doxorubicin has been studied exten
content was not altered in the doxorubicin 1-day group, there sively by the morphological approach. Mettler et al. (20) re-
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 PHOSPHOLIPASE
a acid from L-a-phosphalidylcholine
26—
no-0I1H1400^
24I22IM116M12108g9
e—
-o11¡IIf
200-11II1_Lcontrol
with ascorbate
ascorbateand
andferrous
control
sulfate(8) sulfate
(8) (8)
Fig. 4. EfTectof incubation of mitochondria with ascorbate and ferrous sulfate.
Both lipid peroxides content and phospholipase activity were increased signifi
cantly compared with control (P < 0.01). Bars, SD; numbers in parentheses,
number of samples.
ported that histological changes in doxorubicin-treated rats
consisted of myocyte vacuolation and degeneration, interstitial
edema, and mild fibroplasia, and that these changes were com
monly observed in other species. These findings almost agree
with those of other reports. On the contrary, the biochemical
approach has not yet fully clarified the mechanism of doxorub-
k-m induced cardiomyopathy. In the present study, rat heart
mitochondria were used for analyzing the mechanism of cardi -
otoxicity. Mitochondria play a fundamental role as an energy-
generating site in cells through oxidative phosphorylation. Tak
ing this important function of mitochondria into consideration,
it is probable that their dysfunction would directly lead to
cardiac dysfunction. In our study, the doxorubicin 4-day group
showed a significant inhibition of the activity of NADH-cyto-
chrome c reducÃ-ase and cytochrome c oxidase. Mailer and
Petering (21) reported an inhibition of oxidative phosphoryla
tion in bovine heart mitochondria by doxorubicin. Ogura et al.
(22) also demonstrated that oxidative phosphorylation in rat
heart mitochondria was inhibited and the lipid peroxide level
in mitochondria was increased. Since mitochondria! phospho-
lipids are essential components in maintaining electron-trans
port activity (23, 24), peroxidation of mitochondrial phospho-
lipids has been believed to have a primary importance in the
genesis of doxorubicin-induced cardiotoxicity. In the present
study, the doxorubicin 4-day group showed a high tissue dox
orubicin concentration and a significant elevation of lipid per
oxide level, although the doxorubicin 1-day group showed a
low tissue doxorubicin concentration and no significant eleva
tion of lipid peroxide level. Accordingly, lipid peroxidation
might be linked with the genesis of doxorubicin-induced mito
chondrial dysfunction. On the contrary, many investigators
emphasized recently that the enhanced activation of phospho
lipase induced various pathological conditions associated with
the breakdown of membrane phospholipids (25-28). In our in
vitro experiment 1, we obtained the result that the disturbance
of the respiratory chain induced by phospholipase occurred
mainly in NADH-cytochrome c reducÃ-ase, and cytochrome c
oxidase as we observed in ihe in vivo study, whereas Yasuda
and Fujita (29) poinled oui lhal Ihe phospholipase lhal mighl
bind lo Ihe membrane slruclures are released by alterations in
the phospholipid structure of the membranes after lipid per
oxidation. In ihe present study, the doxorubicin 1-day group
withoul increasing lipid peroxide conlenl showed no significant
changes of phospholipase activily, while in the doxorubicin 4-
day group, phospholipase activily elevaled remarkably associ
ated with the increase of lipid peroxide conlenl. In our in vitro
experimenl 2, we also confirmed lhal Ihe release of linolenic
Downloaded from cancerres.aacrjournals.org on May 28, 2017. © 1987 American Association for Cancer Research.
AND DOXORUBICIN CARDIOMYOPATHY
dilinolenoyl increased as
sociated wilh the increase of lipid peroxide conlenl. Therefore,
il can be surmised lhal Ihe alterations of membrane phospho
lipid slruclures caused by lipid peroxidalion decrease Ihe re
sistance from Ihe phospholipase attack, resulting in degrading
the mitochondrial membrane. Indeed, the doxorubicin 4-day
group, ihe phospholipid conlenl was decreased. Olson et al.
(30) reported that accelerated calcium influx plays an important
role in ihe genesis of doxorubicin-induced cardiotoxicity. It is
interesling lhal calcium is Ihe essenlial factor in the activation
with of phospholipase. These results indicate that doxorubicin-in
ferrous
(8)
duced mitochondrial dysfunclion is based, al leasl in part, on
Ihe degradalion of milochondrial phospholipids by phospholi
pase, which in turn leads to cardiac dysfunction.
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Role of Phospholipase in the Genesis of Doxorubicin-induced
Cardiomyopathy in Rats
Yuuichi Ogawa, Taizo Kondo, Satoru Sugiyama, et al.

Cancer Res 1987;47:1239-1243.

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