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Effects of YM264, a novel PAF antagonist, on puromycin aminonucleoside-induced nephropathy in the rat
Vol. 176, No. 2, 1991
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Pages 78.1-785
April 30, 1991
EFFECTS OF YM26~, A NOVEL PAFANTAGONIST, ON PUROMYCIN AMINONUCLEOSIDE-INDUCEDNEPHROPATHYINTHERAT Toshimitsu Yamada, Kenichi Tomioka, Mami Horie, Yasuhiro Sakurai*, Hitoshi Nagaoka and Toshiyasu Mase Central Research Laboratories, *Product Development Laboratories, Yamanouchi Pharmaceutical Co., Ltd., 21, Miyukigaoka, Tsukuba, Ibaraki 305, Japan Received February 28, 1991
SUMMARY: We investigated the effects of YM264, WEB2086, methylprednisolone and ticlopidine on puromycin-induced nephropathy in the rat. Puromycin produces marked proteinuria, hypercholesterolemia, and hypoalbuminemia. The structurally differing PAF antagonists YM264 and WEB2086 inhibited proteinuria and improved hypercholesterolemia and hypoalbumlnemia. Methylprednisolone also exhibited a beneficial effect on these variables. However, ticlopidine, a platelet inhibitor, showed no inhibitory effect on nephropathy. These results indicate that PAF may play a major role in puromycin-induced nephropathy in the rat, and that PAF antagonists may prove of therapeutic value in the treatment of nephropathy in humans. © 1991 A c a d e m i c
Press,
Inc.
Platelet-activating
factor
(PAF)
is
a new
class
of phospholipid
mediators of allergic and inflammatory reactions (1,2), and is produced by various
inflammatory
cells
such
macrophages (5) and platelets (6).
as
neutrophils
(3), eosinophils
(4),
It has been recently shown that PAF is
synthesized and released in the kidney (7,8), playing an important role in renal
pathophysiology
(9).
For
example,
PAF
induces
contraction
of
mesangial cells (10), a loss of anionic charge in the glomeruli (11), an enhancement oxygen
of vascular permeability
(13). These properties
renal dysfunction (14). examine
(12), and generation of reactive
are closely related to renal injury and
We used YM264 (15), a novel PAF antagonist,
to
the possible role of PAF in puromycin aminonucleoside-induced
nephropathy
in
rats.
This
nephropathy
is
a model
of minimal-change
glomerulonephritis in humans.
MATEI~_,B AND METHODS
Drugs: Puromycin aminonucleoside (Sigma, MO, U.S.A.) and methylprednisolone sodium succinate (Upjohn, MI, U.S.A.) were obtained commercially. 1-(3-methyl-3-phenylbutyl)-4-[2-(3-pyridyl)thiazolidin-4ylcarbonyl]piperazine fumarate (YM264), WEB2086, and ticlopidine were synthesized in our laboratories.
781
0006-29IX/91 $1.50 Copyright © 1991 by Academic Press, Inc. All rights of reproduction in any form reserved.
Vol. 176, No. 2, 1991
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Animals: Male Wistar rats were obtained from Japan SLC (Hamamatsu, Japan). Methods: Rats weighing 180-210 g received a single ip dose of puromycin aminonucleoside (50 mg/kg). They were then randomly allocated to groups. Rats were individually housed in stainless-metabolic cages and allowed free access to water and food throughout the experimental period. Test compounds were orally dosed once (methylprednisolone and ticlopidine) or twice a day (YM264 and WEB2086) from day 0 (the day of puromycin injection ) up to day 11. Protein concentrations in 24-hour urine samples were measured on days 3, 5, 7, 9 and 11 by the sulfosalicylic acid method. Twelve days after the puromycin injection, rats were sacrificed under anesthesia with chloroform and ether. Blood samples were obtained for the assay of serum cholesterol, creatinine and albumin. These serum parameters were enzymatically measured by Hitachi automatic analyzer 73610 (Hitachi, Japan). The histological studies were performed on renal tissues obtained on day 12. For light microscopy studies, renal tissues were fixed in phosphate-buffered 10% formalin and embedded in paraffin. Sections were then stained with hematoxylin-eosin (HE) or with periodic acid-Schiff (PAS) reagent. Statistics: Statistical analysis was carried out by one-wayANOVA (p<0.05) and significant differences from control (p<0.05) were tested by Fisher's protected least-significant difference procedure.
RESULTS Fig. 1 showed the time course of urinary protein excretion in rats. In control
animals,
increasing excretion
to
protein
excretion
peak on day 9.
by
30
to
50%
began
YM264
of
control
to elevate on day 5, rapidly
dose-dependently
inhibited protein
values
day
(p<0.05,
9).
Animals
receiving methylprednisolone also showed lower protein excretion (Fig. I ). Serum
cholesterol
normal the
(p<0.01),
control
levels
in
control
rats were
three times higher than
and a significant reduction of serum albumin was seen in
group
(p<0.01)(Table
I).
Furthermore,
renal
function
200
/ 150 v
--~-- Normal control Nephrotic control ---(3--YM264 0,3 mg/kg --~-- YM264 3 mg/kg
/ f
,
\
i00
~,
50
0 -
.
I
I
3
6
~
[
I
9
12
Days after the injectionof puromycin
Fig. 1.
Effects of ~264 ~ d met~iprednisolone on urinary protein excretion in nephrotic rats injected with puromycin ~inonucleoside. Each point represents the mean ± SE of results obtained in 5 to 8 rats. *, **; S i ~ i f i c ~ t difference from nephrotic controls at p<0.05 ~ d p<0.Ol, respectively.
782
as
Voli 176, No. 2, 1991
Table 1.
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Effects of YM264 and methylprednisolone on serum parameters in nephrotic r a t s
Compound
Dose N (mg/kg, po)
Normal control Nephrotic control YH264
5 8 8 8 8
--0.3 3 2
~ethylprednisolone
Cholesterol (mg/dl) 42 131 105 90 60
Albumin (g/dl)
± 4** ± 15 ± 16 ± 17 ± 6**
3.20 2.69 2,79 2.90 2.93
--4- 0.05** ± 0.04 ± 0.09 ± 0.10 _+ O. 05
Creatinine (mg/dl) 0.78 0.84 0.81 0.74 O. 76
± _ ± ± ±
0,02 0,03 0.06 0.02 O, 02
YM264 was o r a l l y administered twice daily, and methylprednisolone once d a i l y from the day of puromycin i n j e c t i o n for 12 days. Serum parameters were determined on the 12th day. **: S i g n i f i c a n t l y d i f f e r e n t from nephrotic controls a t p<0.01.
assessed by serum creatinine showed a slight deterioration in the control group, although there was no statistically significant difference between the control and normal tended
to
inhibit
prednisolone
at
cholesterol.
a
groups.
these dose
YM264
changes of
in
at doses of 0.3 and 3.0 mg/kg po a dose-dependent
2 mg/kg po prevented
In histopathological
manner.
Methyl-
the increase
in serum
examinations, widening of the tubular
lumen and the formation of urinary casts were frequently seen; disruption of the tubular epithelium was sporadically found in the puromycin-treated control
group.
absorption
A
slight
droplets were
increase
in mesangial
cells
and PAS-positive
found in many gromeruli of this group.
On the
300 - - - ~ - - Normal control -~
Nepnrotic control
0.2 mg/k
~ - - WEB2086 --[]-- Ticlopidine
5 mg/k I00 mg/k,
--~ Methylpred.
c~
[
--0-- YM264
2 mglki /
/
/
~
'
I
i
t
I
. ~ ~ ~ I ~ b ~
!
~I
200 v
c
o
Z
.~ 100
I
0
3 6 9 Days after the Dljectionof puromYcin
12
Fig. 2. Effects of YM264, WEB2086, ticlopidine, and methylpredniselene on urinary protein excretion in nephrotic rats injected with puremycin aminonucleoside. Each point represents the mean ± SE of results obtained in 4 to 7 rats. *, **; Significant difference from nephrotic controls at p<0.05 and p<0.01, respectively. 783
Vol. 176, N o. 2, 1991
Table 2.
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
E f f e c t s of YM264, WEB2086, t i c l o p i d i n e and methylprednisolone on
serum parameters in nephrotic rats
Compound
N
Cholesterol (mg/dl)
--0.2
4 6 7
5
6
i00 2
6 6
50 150 106 111 272 88
Dose (mg/~, pe)
Normal c o n t r o l
Nephrotic control YM264
WEB2086 Ticlopidine Methylprednisolone
_ ± _ _+ _ ±
Albumin (g/dl)
2 * 8 16 26 23 ** 10
3.08 2.53 2.68 2.78 2.34 2.73
_ _ +_ ± _ ±
0.04 * 0.03 0.10 0.16 0.06 0.06
YM264 andWgB2086 were o r a l l y administered twice daily, and ticlopidineand methylprednisotone once d a i l y from the day of puromycin injection for 12 days. Serum parameters were determined on the 12th day. *, **: Significantly differ-
ent from nephrotic controls at p
other hand,
we
saw no abnormalities
in the urinary tubuli of animals
treated with YM264, except a slight increase in mesangial cells and a few absorption antagonist)
droplets.
We
next
and ticlopidine
examined
(platelet
the
effects
inhibitor),
of WEB2086
(PAF
as well as YM264 and
methylprednisolone, on puromycin-induced nephropathy in rats. YM264 (0.2 mg/kg
po)
and methylprednisolone
(2.0 mg/kg
po)
again
inhibited the
increase in urinary protein excretion by 40 to 60% of control values (Fig. 2), and tended to improve hypercholesterolemia and hypoalbuminemia (Table 2).
WEB2086 at a dose of 5 mg/kg po gave the same beneficial effects as
YM264 and methylprednisolone.
On the other hand, ticlopidine at the dose
of 100 mg/kg po, which is enough to exhibit an anti-platelet action in ex vivo experiments, showed no inhibitory effects on nephrotic symptoms.
DISCUSSION
In
our
study,
puromycin-induced
methylprednisolone nephropathy
in
showed
rats,
an
inhibitory
supporting
the
effect
efficacy
on of
steroids in the treatment of minimal-change glomerulonephritis in humans. Although steroids are primarily used in clinical treatments, the clinical application approaches
is have
limited been
due
to
undertaken
their in
severe the
side
effects.
treatment
of
Differing
these diseases.
Recently, Donadio (16) demonstrated that platelet inhibitors were valuable in the treatment of membrane-proliferative glomerulonephritis in humans. In our experiments, the platelet inhibitor ticlopidine showed no inhibitory effects on proteinuria, hypercholesterolemia, and hypoalbuminemia in rats.
Platelets may have a minor role in this animal model.
On the other
hand, Egido et al.(17) reported that PAF antagonists ginkgolide (BN52021), triazolam,
and
alplazolam
inhibited
the
adriamycin-induced nephropathy in rats. 784
urinary
protein excretion in
Our study showed that the struc-
V o l . 176, No. 2, 1991
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
turally differing PAF-receptor antagonists YM264 and WEB2086 improved the proteinuria, hypercholesterolemia and hypoalbuminemia in puromycin-induced nephropathy.
Note that these favorable effects were obtained at doses
similar to the ED50 values for these drugs against PAF in rats, ie, 0.3 mg/kg po for YM264 and 4.2 mg/kg po for WEB2086 (15).
These data support
that PAF has a major role in the development of nephrotic syndromes. In
summary,
our
study
suggests
puromycin-induced nephropathy
that
PAF
has a specific role in
in rats, and that the PAF antagonist may
prove of therapeutic value in the treatment of nephropathy in humans. ACKNO~.~nGMENTS
The authors wish to thank Drs. H. Maeno, N. Inukai, T. Miki, K. Murase and I. Ohata for their encouragement during this work. REFERENCES
I. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17.
Demopoulos, C.A., Pinckard, R.N.,and Hanahan, D.J. (1979) J. Biol. Chem. 254, 9355-9358. Benveniste, J., Tenee, M., Bidault, J., Boullet, C., Varence, P., and Polonsky, J. (1979) C.R.Seances Acad. Sci., Ser. D289, 1037-1040. Lynch, J.M., Lontner, G.Z., Betz, S.J.,and Henson, P.M. (1979) J. Immunol. 123, 1219-1226. Lee, T., Lenihan, D.J., Malone, B., Roddy, L.L.,and Wasserman, S.I. (1984) J. Biol. Chem. 259, 5526-5530. Albert, D.H.,and Snyder, F. (1983) J. Biol. Chem. 258, 97-102. Chignard, M., Le Couedic, J.P., Tence, M., Vargaftig, B.B.,and Benveniste, J. (1979) Nature (Lend.) 275, 799-800. Schlondorff, D., Goldwasser, P., Neuwirth, R., Satriano, J.A.,and Clay, K.L. (1986) Am. J. Physiol. 250, F1123-F1127. Pirotzky, E., Bidault, J., Burtin, C., Gubler, M.C.,and Benveniste, J. (1984) Kidney Int. 25, 404-410. Camussi, G. (1986) Kidney Int. 29, 469-477. Schlendorff, D., Satriano, J.A., Hagege, J., Perez, J.,and Baud, L. (1984) J. Clin. Invest. 73, 1227-1231. Camussi, G., Tetta, C., Coda, R., Segoloni, G.P.,and Vereellone, A. (1984) Kidney Int. 25, 73-81. O'Flahrty, J.T., Wycle, R.L., Miller, C.H., Lewis, J.C., Waite, M., Bass, D.A., McCall, C.E.,and DeChatelet, L.R. (1981) Am. J. Pathol. 103, 70-79. Sedor, J.R.,and Abboud H.E. (1985) Kidney Int. 27, 222A. Schwertschlag, U., Scherf, H., Gerber, J.G., Mathias, M.,and Nies, A.S. (1987) Circulation Res. 60, 534-539. Yamada, T., Tomioka, K., Saito, M., Horie, M., Mase, T., Hara, H., and Nagaoka, H. (1991) Arch. Int. Pharmaeodyn. Therapie, In press. Donadie, J.V. (1984) New Engl. J. Med. 310, 1421-1426. Egido, J., Robles, A., 0rtiz, A., Ramirez, F., Gonzalez, E., Mampaso, F., Crespo, M.S., Braquet, P.,and Hernando, L. (1987) Eur. J. Pharmacol. 138, 119-123.
785
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Pages 78.1-785
April 30, 1991
EFFECTS OF YM26~, A NOVEL PAFANTAGONIST, ON PUROMYCIN AMINONUCLEOSIDE-INDUCEDNEPHROPATHYINTHERAT Toshimitsu Yamada, Kenichi Tomioka, Mami Horie, Yasuhiro Sakurai*, Hitoshi Nagaoka and Toshiyasu Mase Central Research Laboratories, *Product Development Laboratories, Yamanouchi Pharmaceutical Co., Ltd., 21, Miyukigaoka, Tsukuba, Ibaraki 305, Japan Received February 28, 1991
SUMMARY: We investigated the effects of YM264, WEB2086, methylprednisolone and ticlopidine on puromycin-induced nephropathy in the rat. Puromycin produces marked proteinuria, hypercholesterolemia, and hypoalbuminemia. The structurally differing PAF antagonists YM264 and WEB2086 inhibited proteinuria and improved hypercholesterolemia and hypoalbumlnemia. Methylprednisolone also exhibited a beneficial effect on these variables. However, ticlopidine, a platelet inhibitor, showed no inhibitory effect on nephropathy. These results indicate that PAF may play a major role in puromycin-induced nephropathy in the rat, and that PAF antagonists may prove of therapeutic value in the treatment of nephropathy in humans. © 1991 A c a d e m i c
Press,
Inc.
Platelet-activating
factor
(PAF)
is
a new
class
of phospholipid
mediators of allergic and inflammatory reactions (1,2), and is produced by various
inflammatory
cells
such
macrophages (5) and platelets (6).
as
neutrophils
(3), eosinophils
(4),
It has been recently shown that PAF is
synthesized and released in the kidney (7,8), playing an important role in renal
pathophysiology
(9).
For
example,
PAF
induces
contraction
of
mesangial cells (10), a loss of anionic charge in the glomeruli (11), an enhancement oxygen
of vascular permeability
(13). These properties
renal dysfunction (14). examine
(12), and generation of reactive
are closely related to renal injury and
We used YM264 (15), a novel PAF antagonist,
to
the possible role of PAF in puromycin aminonucleoside-induced
nephropathy
in
rats.
This
nephropathy
is
a model
of minimal-change
glomerulonephritis in humans.
MATEI~_,B AND METHODS
Drugs: Puromycin aminonucleoside (Sigma, MO, U.S.A.) and methylprednisolone sodium succinate (Upjohn, MI, U.S.A.) were obtained commercially. 1-(3-methyl-3-phenylbutyl)-4-[2-(3-pyridyl)thiazolidin-4ylcarbonyl]piperazine fumarate (YM264), WEB2086, and ticlopidine were synthesized in our laboratories.
781
0006-29IX/91 $1.50 Copyright © 1991 by Academic Press, Inc. All rights of reproduction in any form reserved.
Vol. 176, No. 2, 1991
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Animals: Male Wistar rats were obtained from Japan SLC (Hamamatsu, Japan). Methods: Rats weighing 180-210 g received a single ip dose of puromycin aminonucleoside (50 mg/kg). They were then randomly allocated to groups. Rats were individually housed in stainless-metabolic cages and allowed free access to water and food throughout the experimental period. Test compounds were orally dosed once (methylprednisolone and ticlopidine) or twice a day (YM264 and WEB2086) from day 0 (the day of puromycin injection ) up to day 11. Protein concentrations in 24-hour urine samples were measured on days 3, 5, 7, 9 and 11 by the sulfosalicylic acid method. Twelve days after the puromycin injection, rats were sacrificed under anesthesia with chloroform and ether. Blood samples were obtained for the assay of serum cholesterol, creatinine and albumin. These serum parameters were enzymatically measured by Hitachi automatic analyzer 73610 (Hitachi, Japan). The histological studies were performed on renal tissues obtained on day 12. For light microscopy studies, renal tissues were fixed in phosphate-buffered 10% formalin and embedded in paraffin. Sections were then stained with hematoxylin-eosin (HE) or with periodic acid-Schiff (PAS) reagent. Statistics: Statistical analysis was carried out by one-wayANOVA (p<0.05) and significant differences from control (p<0.05) were tested by Fisher's protected least-significant difference procedure.
RESULTS Fig. 1 showed the time course of urinary protein excretion in rats. In control
animals,
increasing excretion
to
protein
excretion
peak on day 9.
by
30
to
50%
began
YM264
of
control
to elevate on day 5, rapidly
dose-dependently
inhibited protein
values
day
(p<0.05,
9).
Animals
receiving methylprednisolone also showed lower protein excretion (Fig. I ). Serum
cholesterol
normal the
(p<0.01),
control
levels
in
control
rats were
three times higher than
and a significant reduction of serum albumin was seen in
group
(p<0.01)(Table
I).
Furthermore,
renal
function
200
/ 150 v
--~-- Normal control Nephrotic control ---(3--YM264 0,3 mg/kg --~-- YM264 3 mg/kg
/ f
,
\
i00
~,
50
0 -
.
I
I
3
6
~
[
I
9
12
Days after the injectionof puromycin
Fig. 1.
Effects of ~264 ~ d met~iprednisolone on urinary protein excretion in nephrotic rats injected with puromycin ~inonucleoside. Each point represents the mean ± SE of results obtained in 5 to 8 rats. *, **; S i ~ i f i c ~ t difference from nephrotic controls at p<0.05 ~ d p<0.Ol, respectively.
782
as
Voli 176, No. 2, 1991
Table 1.
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
Effects of YM264 and methylprednisolone on serum parameters in nephrotic r a t s
Compound
Dose N (mg/kg, po)
Normal control Nephrotic control YH264
5 8 8 8 8
--0.3 3 2
~ethylprednisolone
Cholesterol (mg/dl) 42 131 105 90 60
Albumin (g/dl)
± 4** ± 15 ± 16 ± 17 ± 6**
3.20 2.69 2,79 2.90 2.93
--4- 0.05** ± 0.04 ± 0.09 ± 0.10 _+ O. 05
Creatinine (mg/dl) 0.78 0.84 0.81 0.74 O. 76
± _ ± ± ±
0,02 0,03 0.06 0.02 O, 02
YM264 was o r a l l y administered twice daily, and methylprednisolone once d a i l y from the day of puromycin i n j e c t i o n for 12 days. Serum parameters were determined on the 12th day. **: S i g n i f i c a n t l y d i f f e r e n t from nephrotic controls a t p<0.01.
assessed by serum creatinine showed a slight deterioration in the control group, although there was no statistically significant difference between the control and normal tended
to
inhibit
prednisolone
at
cholesterol.
a
groups.
these dose
YM264
changes of
in
at doses of 0.3 and 3.0 mg/kg po a dose-dependent
2 mg/kg po prevented
In histopathological
manner.
Methyl-
the increase
in serum
examinations, widening of the tubular
lumen and the formation of urinary casts were frequently seen; disruption of the tubular epithelium was sporadically found in the puromycin-treated control
group.
absorption
A
slight
droplets were
increase
in mesangial
cells
and PAS-positive
found in many gromeruli of this group.
On the
300 - - - ~ - - Normal control -~
Nepnrotic control
0.2 mg/k
~ - - WEB2086 --[]-- Ticlopidine
5 mg/k I00 mg/k,
--~ Methylpred.
c~
[
--0-- YM264
2 mglki /
/
/
~
'
I
i
t
I
. ~ ~ ~ I ~ b ~
!
~I
200 v
c
o
Z
.~ 100
I
0
3 6 9 Days after the Dljectionof puromYcin
12
Fig. 2. Effects of YM264, WEB2086, ticlopidine, and methylpredniselene on urinary protein excretion in nephrotic rats injected with puremycin aminonucleoside. Each point represents the mean ± SE of results obtained in 4 to 7 rats. *, **; Significant difference from nephrotic controls at p<0.05 and p<0.01, respectively. 783
Vol. 176, N o. 2, 1991
Table 2.
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
E f f e c t s of YM264, WEB2086, t i c l o p i d i n e and methylprednisolone on
serum parameters in nephrotic rats
Compound
N
Cholesterol (mg/dl)
--0.2
4 6 7
5
6
i00 2
6 6
50 150 106 111 272 88
Dose (mg/~, pe)
Normal c o n t r o l
Nephrotic control YM264
WEB2086 Ticlopidine Methylprednisolone
_ ± _ _+ _ ±
Albumin (g/dl)
2 * 8 16 26 23 ** 10
3.08 2.53 2.68 2.78 2.34 2.73
_ _ +_ ± _ ±
0.04 * 0.03 0.10 0.16 0.06 0.06
YM264 andWgB2086 were o r a l l y administered twice daily, and ticlopidineand methylprednisotone once d a i l y from the day of puromycin injection for 12 days. Serum parameters were determined on the 12th day. *, **: Significantly differ-
ent from nephrotic controls at p
other hand,
we
saw no abnormalities
in the urinary tubuli of animals
treated with YM264, except a slight increase in mesangial cells and a few absorption antagonist)
droplets.
We
next
and ticlopidine
examined
(platelet
the
effects
inhibitor),
of WEB2086
(PAF
as well as YM264 and
methylprednisolone, on puromycin-induced nephropathy in rats. YM264 (0.2 mg/kg
po)
and methylprednisolone
(2.0 mg/kg
po)
again
inhibited the
increase in urinary protein excretion by 40 to 60% of control values (Fig. 2), and tended to improve hypercholesterolemia and hypoalbuminemia (Table 2).
WEB2086 at a dose of 5 mg/kg po gave the same beneficial effects as
YM264 and methylprednisolone.
On the other hand, ticlopidine at the dose
of 100 mg/kg po, which is enough to exhibit an anti-platelet action in ex vivo experiments, showed no inhibitory effects on nephrotic symptoms.
DISCUSSION
In
our
study,
puromycin-induced
methylprednisolone nephropathy
in
showed
rats,
an
inhibitory
supporting
the
effect
efficacy
on of
steroids in the treatment of minimal-change glomerulonephritis in humans. Although steroids are primarily used in clinical treatments, the clinical application approaches
is have
limited been
due
to
undertaken
their in
severe the
side
effects.
treatment
of
Differing
these diseases.
Recently, Donadio (16) demonstrated that platelet inhibitors were valuable in the treatment of membrane-proliferative glomerulonephritis in humans. In our experiments, the platelet inhibitor ticlopidine showed no inhibitory effects on proteinuria, hypercholesterolemia, and hypoalbuminemia in rats.
Platelets may have a minor role in this animal model.
On the other
hand, Egido et al.(17) reported that PAF antagonists ginkgolide (BN52021), triazolam,
and
alplazolam
inhibited
the
adriamycin-induced nephropathy in rats. 784
urinary
protein excretion in
Our study showed that the struc-
V o l . 176, No. 2, 1991
BIOCHEMICAL AND BIOPHYSICAL RESEARCH COMMUNICATIONS
turally differing PAF-receptor antagonists YM264 and WEB2086 improved the proteinuria, hypercholesterolemia and hypoalbuminemia in puromycin-induced nephropathy.
Note that these favorable effects were obtained at doses
similar to the ED50 values for these drugs against PAF in rats, ie, 0.3 mg/kg po for YM264 and 4.2 mg/kg po for WEB2086 (15).
These data support
that PAF has a major role in the development of nephrotic syndromes. In
summary,
our
study
suggests
puromycin-induced nephropathy
that
PAF
has a specific role in
in rats, and that the PAF antagonist may
prove of therapeutic value in the treatment of nephropathy in humans. ACKNO~.~nGMENTS
The authors wish to thank Drs. H. Maeno, N. Inukai, T. Miki, K. Murase and I. Ohata for their encouragement during this work. REFERENCES
I. 2. 3. 4. 5. 6. 7. 8. 9. 10. 11. 12. 13. 14. 15. 16. 17.
Demopoulos, C.A., Pinckard, R.N.,and Hanahan, D.J. (1979) J. Biol. Chem. 254, 9355-9358. Benveniste, J., Tenee, M., Bidault, J., Boullet, C., Varence, P., and Polonsky, J. (1979) C.R.Seances Acad. Sci., Ser. D289, 1037-1040. Lynch, J.M., Lontner, G.Z., Betz, S.J.,and Henson, P.M. (1979) J. Immunol. 123, 1219-1226. Lee, T., Lenihan, D.J., Malone, B., Roddy, L.L.,and Wasserman, S.I. (1984) J. Biol. Chem. 259, 5526-5530. Albert, D.H.,and Snyder, F. (1983) J. Biol. Chem. 258, 97-102. Chignard, M., Le Couedic, J.P., Tence, M., Vargaftig, B.B.,and Benveniste, J. (1979) Nature (Lend.) 275, 799-800. Schlondorff, D., Goldwasser, P., Neuwirth, R., Satriano, J.A.,and Clay, K.L. (1986) Am. J. Physiol. 250, F1123-F1127. Pirotzky, E., Bidault, J., Burtin, C., Gubler, M.C.,and Benveniste, J. (1984) Kidney Int. 25, 404-410. Camussi, G. (1986) Kidney Int. 29, 469-477. Schlendorff, D., Satriano, J.A., Hagege, J., Perez, J.,and Baud, L. (1984) J. Clin. Invest. 73, 1227-1231. Camussi, G., Tetta, C., Coda, R., Segoloni, G.P.,and Vereellone, A. (1984) Kidney Int. 25, 73-81. O'Flahrty, J.T., Wycle, R.L., Miller, C.H., Lewis, J.C., Waite, M., Bass, D.A., McCall, C.E.,and DeChatelet, L.R. (1981) Am. J. Pathol. 103, 70-79. Sedor, J.R.,and Abboud H.E. (1985) Kidney Int. 27, 222A. Schwertschlag, U., Scherf, H., Gerber, J.G., Mathias, M.,and Nies, A.S. (1987) Circulation Res. 60, 534-539. Yamada, T., Tomioka, K., Saito, M., Horie, M., Mase, T., Hara, H., and Nagaoka, H. (1991) Arch. Int. Pharmaeodyn. Therapie, In press. Donadie, J.V. (1984) New Engl. J. Med. 310, 1421-1426. Egido, J., Robles, A., 0rtiz, A., Ramirez, F., Gonzalez, E., Mampaso, F., Crespo, M.S., Braquet, P.,and Hernando, L. (1987) Eur. J. Pharmacol. 138, 119-123.
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