1. No category

319
ῌ
Vol. 34, pp. 319333, 2006
ῐ
UUO Eplerenone Ep AT1a AT1a 1
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1
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Ang II !"#$% AT1a "&'()*+,-./0123456789
% ':;<=>?9%@A Ang II !BCD,EFGHI:% Ang II " AT1a
JKL* AT1a >:I ()*+,"M Eplerenone Ep 6N%OP
/Q Unilateral Ureteral Obstruction RS( UUO !./012345TB>
UVL* AT1a 'WXYZX@ AT1a n40 ' AT1a n41 >
[H\H Ep ] Ep 200 mgkgday EP ^' Ep ] Ep ^6_`@ Ep ] 7 a6b/QRS( UUO >BX UUO B 246 a6cd>efX cgh23
!ij>kX@ AT1a AT1a !^# Ep ]6N%lmnopq-
ArHmst@ AT1a ! Ep ^#- Ep ^6u vwxy type I ' type IV
!zCl6TGH {|}#kX@./0123~l6GH@€'s
r Ep - 01‚ƒ!45>T9%'„…rH@ AT1a ! Ep ^#- AT1a
![H6u MCP-1 †z vwxy type I ' type IV zCl6TGH
./0123!ij~‡:ˆ‰#$t@ XsX AT1a ! Ep ^' Ep ^#- ./0123CŠij#$‹ Ep ]6N%Grm%./0123TŒ- ArH
mst@
Q'XI Ep - UUO 6Ž`%./0123!45>T9%CGH@ AT1a
#- Ep !ŒCArHmst@N‹ UUO 6Ž`% Ep !01‚ƒT-
AT1a "sr!‚ƒ‘4’“(>”•9%€'6N‹U–%—˜C™GH@
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Unilateral Ureteral Obstruction UUO ./0123
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12῍ 0.2 g 5/῍ 0€ Ep  Ep ῍ AT1
ῌ a ; n20, AT1a ; n21 Ep ;
῍ ; Ep ῍ AT1a ; n20, AT1a ; n
!"
Ang II#35῍ Ang II $
20 <4ῌ Ep 1 0.5 Carboxymethyl
!"
%& ACEI '(
Cellulose Sodium Salt CMC῍ =‚ƒ„…†‡῍
) AngII *+ ARB ῍ >ˆ ?‰4 Ep 100 mg kg day῍ rsŠ‹C
,-./68ῌ „…†‡Œ 5῍ ; Ep 1 0῎5 CMC
ῌ
῍ 0$1῍ 21//
5 1 @ 2 Ž]῍ UUO $A
48
ACEI 341 ARB 5678/
BC‘?34ῌ D1EF’'
9:;<!=῍ >?
/῍ no<]13#.'\῍
/@:;<!=A<BCDE9 -.῍
xI“]D5”•G4ῌ Ep –
FG῍ :;<!=$5H=IJ?
— 7 @˜῍ ™HšC: 55 mgkg 5H›I῍
:;<!=*+
J@KL„…†‡῍ Jœ MNž5 2 Ÿ
1011
ῌ :;
OV$῍ 05PQ῍ UUO ST:5$A
!"-:;<!
17῍ UUO $A 2῍ 4῍ 6 @˜ UUO-day2῍4῍6 OPQ!R῍ ž B5C‘4ῌ 71 Satoh4 # ῍ :;<!=$5
AT1a no< UUO ST:'/῍ day5
,/@K.4
<!=1῍ LM-
=N RAAS 12
H=IJ?:;<!=*+1῍ 2bc.῍ day10 '/.
5?ῌ
¡4e(#῍ day4῍ 6 ^R
"
ST:U#V$
c5-?4l day2῍4῍6 SG4ῌ <
Unilateral Ureteral Obstruction ST: UUO W
4 Ep  AT1a ; n20, AT1a ; n
XYZ5*[\῍ 5
21 ; Ep  AT1a ; n20῍ AT1a ; n
]^_`aST:13 '/῍ :;<
20 5¢ day2῍4῍6 ῍ Ep  AT1a
!=*+ UUO %bc5
: day2; n6, day4; n7, day6; n7, AT1a 4K.14῍ :;<!=*+
: day2; n7῍ day4; n7, day6; n7, ; Ep 
῍ ,d
AT1a : day2; n6, day4; n7, day6; n7,
.4ῌ 0e῍ f4g1῍ Ang II *
AT1a :day2; n7, day4; n7, day6; n6
AT1a *h&:;<!=1
[4ῌ B£#1῍ w“=C:
ij?ῌ /@k'544ῌ e
/4ῌ Ep ;῍ –HTUno<
k'5()?4l῍ Ang II * AT1a
1῍ –HTU 6 @˜B5C‘.῍ sham op-
m*no< AT1a 5/῍ :;<!=
eration 4ῌ
*+ Eplerenone Ep ( UUO K¤¥1῍ Vn¦§¨©Jª«¬W!­®
$5῍ p+,q
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nJ=rsC-t῍ 0./0E
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monocyte chemoattractant protein-1 MCP-1 5῍
03009, TG050512-1῍ Vn¦§¨©JªW!
u1῍ 2bc3455v?4lw
«¬´Š;xŠµ/¶4ῌ
xCy0E55v῍ 67zno<
AT1a Ep –— Ep –— 7 @˜ UUO $
8{6|94ῌ
A῍ BC‘ tail-cu# ^ BP-98A; ·r“=
„…†‡῍ >ˆ 9¸5_G418ῌ
Ald UUO C 57 BL 6 } I J ~ x o ; 9`:;<!=1῍ RIA-+^_G.
< AT 1 a
15῍16 n40; 8 :῍ 25.90.2 g C57
4 ¹acª BCL „…†‡῍ >ˆῌ
BL 6
monocyte chemoattractant protein-1
< AT1a n41; 8 :῍ 26.3
108
*>/m)¹nkº»¼
321
MCP-1 }~$VW!†‡ˆ|‰Š‹Œ
lysis bu#er 0.1 mol L $ῌ PQRVWŽ'῍ \]^_K8|
῍ 1 mgml ῍ 1 mgml z'(῍ a>8|z
!†
῍ 1 Triton X100῍ 0.05 mmolL phenyl-
‡ˆ UUO 8|
9M:;$ῌ
methyl sulfonyl fluoride ῍ ῍
MCP-1 ELISA
mouse JEMCP-1 im-
munoassay; R&D systems῍ MN῍ USA !"#
$ ῌ %&'(῍ Lowry
‘’(“”•–—˜™ meanSE 8š
DC )
$ῌ 2 ›V9œ( Mann-Whitney U #
*+,-ῌ./.0.1
$ῌ žŸ
5 Z¡¢KL!£™
17
2345῍ 678"#$ῌ MCP-1 gM$ῌ ¡¢wxy
!( SPSS ver. 14.0J
(῍ MCP-1 &'%&'98:;$ῌ
SPSS JAPAN Inc., 67 $ῌ
<$(῍ =>?*@#A.
BCD
E῍ 12 mm FD!GHDE$ῌ IJKL:;M
῍ PAS NOPQRST:;1819῍ *UN
Ep ¤¥¦§¨©ª(῍ AT1a hlm8
OVWSX:; YZ$ῌ [$\]^_K
( AT1a hlm!9«£!¬•8­Z$
LSAB
; `aῌbcd2345῍ 7
p0.001 Table 1ῌ Ep ¤!fg AT1a h
e !fghi
j.khlm F480
lm8©ª¬®(¯°±Z$ῌ AT1a hl
kn; Bio Medicals AG, Augst, Switzerland῍ a
m8(῍ UUO-day4-Ep ›8῍ Ep ¤¥!9«῍
.o type IV .pkhlma.o
¦§¨©ª²£™(±²῍ ¬³´!­Z$ῌ
type IV kn; Southern Biotechnology Associates
17
!"#
Int., Ontario, Canada, a.o type IV .p
Ep ¤ sham ope AT1a M sham ope
khlma.o type IV kn; Southern Bio-
AT1a 8(῍ ©µ*>/m)&'(¶
technology Associates Int., Ontario, Canada '8­Z$AT1a : 39῎0 ngdl῍ AT1a :
:;$ῌ .`q! 5 rst῍ uvw
33.1 ngdl Fig 1ῌ AT1a M AT1a !
xy
Image Analyzer῍ .=him)
„῍ ©µ*>/m)&'( UUO-· Ep ›
q12345῍ 67 ῍ PQRSTz῍
M sham ope 8¶'8­Z$ῌ [$ AT1a VWSXz῍ \]^_K!fZ{E$|
UUO-Ep ›8(῍ £™(±²῍ ©µ*>/
z῍ }~$VW€#$417ῌ a
m)&'( UUO-· Ep ›!9«¸³´!
>‚ƒ„f… UUO !„῍
­Z$ῌ AT1a UUO-day2῍6-Ep ›8(῍
Table 1. Systolic Blood Pressures of Angiotensin II Type 1a Receptor Knockout Mouse
AT1a and AT1a before Administration of Eplerenone Ep, and on
days 0, 2, 4, and 6 after Unilateral Ureteral Obstruction UUO. mmHg
109
KLMN
322
OPῌQRST
Figure 1. Levels of serum aldosterone in AngiotensinIItype 1a receptor AT1a and AT1a . Values are the
meanSE. , sham operated mice of AT1a without administration of Eplerenone Ep; , AT1a with unilateral ureteral obstruction UUO and without administration of Ep; , AT1a with
UUO and administration of Ep; , sham operated mice of AT1a without administration of Ep; , AT
1a with UUO and without administration of Ep; , AT1a with UUO and administration of Ep. p0.05 compared to the AT1a with UUO and without administration of Ep, p0.05 compared to
the AT1a with UUO and without administration of Ep.
UUO-day2῍6- Ep .ῌ
p0.05ῌ AT1 Collagen type I UUO-day2῍6- Ep AT1a ῍ AT1 + day6 day
῍ AT1a ῍ 4 collagen type I p0.05ῌ
ῌ AT1a UUO-day6- Ep MCP-1 ῍ AT1a UUO-day6- Ep ῍ AT1a UUO-day2- Ep ῍ AT1a
HI'
EFG5-.
῍ Collagen type I J'-.
UUO-day2- Ep MCP-
Fig 4ῌ AT1a UUO-day6-Ep 1 !"# p0.05 Fig 2ῌ AT1a ῍ UUO-day6- Ep $ AT1a UUO %&'(῍ Ep $
!"# p0.05ῌ AT1a &'(῍ Ep
Ep MCP-1 )*ῌ UUO-day
$ Ep )*ῌ
4῍6 &'( Ep ῍ Ep + MCP-1 +
Collagen type IV AT1a ῍ AT1 + day6 day
day2 ,'-./῍ 012$3(
AT1a AT1a 4
!"#5
2῍ 4 collagen type IV EFG5
-.ῌ
-.ῌ AT1a UUO-day4- Ep ῍ AT1a UUO-day4- Ep AT1a UUO-day6- Ep ῍ AT1a
6789:;<=
Collagen type IV !"# p
0.05 Fig 5ῌ AT1a UUO-day6-Ep !"# Fig 3ῌ AT1a $ AT1a ῍ UUO-day6- Ep > UUO %&'(῍ Ep $ Ep !"# p0.05ῌ AT1a ῍ Ep $
?@A6789:;<=)*ῌ 0
Ep )*ῌ
AT1a ῍ AT1a + day6 &'(
AT1a ῍ AT1a + day4῍6 day
day2῍ 4 6789:;<=BCD5110
*9:;<=,qrs>?@¨©LI
323
Figure 2. Protein expressions of monocyte chemoattractant protein- 1 ῌMCP-1῍ in AngiotensinIItype 1 a receptor
῎AT1a῏ ῌῐΐῐ῍ and AT1a ῌ῕ΐ῕῍. The expression of MCP-1 protein was determined by ELISA and was
corrected for the total amount of protein. Values are the meansῑSE. , AT1a ῌῐΐῐ῍ with unilateral
ureteral obstruction ῌUUO῍ and without administration of Eplerenone ῌEp῍; , AT1a ῌῐΐῐ῍ with UUO and
administration of Ep; , AT1a ῌ῕ΐ῕῍ with UUO and without administration of Ep; , AT1a ῌ῕ΐ῕῍ with UUO
and administration of Ep. ῔pῒ0.05 compared to the AT1a ῌῐΐῐ῍ with UUO and without administration of
Ep on day 2 after UUO.
2 ῍ #fgYZῌ Ep G
ῌ PAS ῍
῍ hi
AT1a ῌ῕ΐ῕῍ AT1a ῌῐΐῐ῍ UUO MCP-1 jk`a῍ \]^_, type I type IV `
῍ Ep
a2/0CDlZ῍ -.
m3
!"#$%&'() ῌFig 6῍ῌ
!T( AT1a ῌ῕ΐ῕῍
!T( AT1a ῌῐΐῐ῍ ῍
*+,-.
῍ AT1a
ῌ )῍ AT1a ῌ῕ΐ῕῍ [T῍ Ep
ῌῐΐῐ῍ UUO ῍ AT1a ῌ῕ΐ῕῍ UUO !"#-.
no&'YZ()
/ 0 1 ( 2῍ 3 ῌFig 7῍ῌ
ῌ *9:;<=,["p AT1a qrsGt
AT1a ῌῐΐῐ῍ UUO-day 6-Ep 4῍ UUO-day
AngII u=vw῍ xZyZ-.
GC
6-5 Ep 4-.
῍ /03
D2῍ Xz{|}CDEF῍ &'YZ(
)ῌpῒ0.05῍ῌ AT1a ῌ῕ΐ῕῍ UUO ῍ Ep
)ῌ ~ZYF)Y῍ UUO [€# Ep !"#-.
$%&'()ῌ
ῌ
-.bc%CD῍ AT1a qrsGtc%‚
AS!e#ƒ„…9†‡Gˆ‰e#~"T
῍
678῍ *9:;<=,>?@ ῌEp῍
Šh#‹Œ2fgYZ῍ Ž AT1a qrs`
!
a(‘NOTi’“2#fgYZ
"#-.
ABCDEFG AT1a ῌ῕ΐ῕῍῍
ῌ
AT1a ῌῐΐῐ῍ GHI UUO JK9GLMNO
ῌ Ep
QBW”ˆ•–iS!T# AngII
2῍ QB["p5QB—˜}
G™še#~
!PQRST῍ AT1a ῌῐΐῐ῍
AT1a ῌ῕ΐ῕῍ /0UV2῍ Ep
῍ ›iœYZT#ῌ AngIIqrs῍ 1 
!"#/0(QR$W῍ AT1a ῌ῕ΐ῕῍῍ AT1a ῌῐ
ῌAT1῍ 2  ῌAT2῍ 2žŸ῍ AT1 2 ¡
ΐῐ῍ X4&'YZ()ῌ AT1a ῌῐΐῐ῍ [
u¢£¤ ῌAT1a AT1b῍ 2#20῍ῌ Satoh Y
T῍ Ep
!"῍ -.\]^_, type I AT1a ῌ῕ΐ῕῍ GI UUO ῍ UUO 5 ¥¦
type IV `aCD῍ -.bc%CD2&
UUO -.bc%CDEF2&'YZ~)
'YZd)Y῍ Ep -.bc%ABGCDe
Y῍ UUO [€#§-.bc%῍ AT1a G
111
Average number of F4/80 positive cells
Per field in the interstitium
324
ῌ
#
30
25
20
15
10
5
0
Day 2 Day 4 Day 6
Ep (-)
Day 2 Day 4 Day 6
Ep (+)
AT1a ( +/+) with UUO
Day 2 Day 4 Day 6
Ep (-)
Day 2
Day 4
Day 6
Ep (+)
AT1a ( -/-) with UUO
Figure 3. Immunostaining of the tubulointerstitium with F 4 80 in the kidneys of AngiotensinIItype 1 a receptor
AT1a and AT1a . F480 immunostaining on day 6 after UUO in the UUO-kidney of AT1a without administration of Eplerenone Ep a, the contralateral non-obstructed kidney of AT1a without administration of Ep b, the unilateral ureteral obstruction UUO -kidney of AT1a with
administration of Ep c, the contralateral non-obstructed kidney of AT1a with administration of Ep
d, the UUO-kidney of AT1a without administration of Ep e, the contralateral non-obstructed
kidney of AT1a without administration of Ep f, the UUO-kidney of AT1a with administration
of Ep g, the contralateral non-obstructed kidney of AT1a with administration of Ep h. i: The
number of F 4 80 -positive cells that had infiltrated the interstitium was assessed semiquantitatively as
described under Materials and Methods. Values are the meanSE. , AT1a with UUO and without
administration of Ep; , AT1a with UUO and administration of Ep; , AT1a with UUO and
without administration of Ep; , AT1a with UUO and administration of Ep. p
0.05 compared to
the AT1a with UUO and without administration of Ep. Original magnifications, 200.
112
)*+,-./6%d&
_¼
325
AngII ῌAngIIΐAT1a῍ pA῍ AT1a 6"
ῌ ῍ UUO 10 ##O{|}*01"23O?&
῍ AngIIΐAT1
2>
XY[~ ῌFig 8῍ῌ ?4
a 
῍ r in vitro €4t-O56=
4῍
! UUO "#$
ῌ
%
&'(4῍ῌ
)*+,-./῍ AngII0123 AT1῍ AT2
.uvwx7῍ .uv8&
"
45ῌ21῎22῎ῌ AT1a
49‚ῌ .uv87῍ O:
ῌῒῑῒ῍ 4῍ AT1a 67
῍ AT2
ƒk
:74=>῍ .v@;2
"
)*+,-./&'(
.uv„,T`C… ,T`,:74=>
ῌ 8)*+,-./῍ Ep &'(ῌ ῍ AZAN <†῍ ‡
9 UUO AT1a ῌῒῑῒ῍ 4῍ Ep oKˆ/ type I k2y type IV =‰<†239
9 UUO AT1a ῌ῏ῑ῏῍ :;<
‚ῌ ‡oKˆ/ IῐX Q4Š‹W>
4 = 3῍ Ep UUO k3῍ e?!@ 1000 ŒŽ)n_9
21῎
AT1a ῌῒῑῒ῍ 4῍ Ep 9 UUO W…+‘’s3 3 “AB”C"uD
AT1a ῌῒῑῒ῍ :;<4=>ῌ ?@
•4῍ type I ?–Q3E— 10ῐ80 nm
A῍ UUO AT1a ῌῒῑῒ῍ 4῍ AT1a 6"
7uF"9῍ ˜?GM–™d
AngIIB.CD
?&4῍
šF"›H῍ I῍ œ9ŽJ
AT1a 6EF9῍ GH-IBJIK
ῌ žK type IV L‡oKˆ/&Ÿ ῍ M
+LCD@A῍ AngII5MN῍ AT1a ῌ῏ῑ
.uveNL¡69”HH4=ῌ type I F
῏῍ : AngII ῍ = AngII O6
day6 E4O¢
ql
P6<Q3῍ )*+,-.
&'(ῌ type IV eNL¡6P£
23῍
/5ῌ<Q῍ RS=&'
῍ Q¤23FO¢
&'(῍
(῍ UUO AT1a ῌῒῑῒ῍ 4῍ )*+,T./
AT1a ῌῒῑῒ῍ ῍ ¥
O
R¦
M4
U,VKWX
Y=ῌ Q UUO
"#O?&῍ .v@;
9§S
AT1a ῌῒῑῒ῍ 4῍ Ep 4)*+,-./
7O UUO 7
B.CD῍
49‚ῌ rsij* UUO 4῍ MCP-
Z!)*+,-./
1 O¢῍ 8 day4 S4῍ R
5ῌ"#Y[ῌ
¨qT
day6 4 MCP-1 O¢2>
)*+,-./῍ \]ῌ##^_,
©D.JªK«¬­῍ F"Xῌ
T`,"$aO?&4
bc"#$%῍ )*+
,-./6%d&R"῍ U
VW^X.Y
]c®"¯Z°±
bc#$"O῍ '(9e)fg4*h
²4῍ ACEI l ARB &῍ )*+,-./
24ῒ28῎ῌ UUO ij*k
l῍ )*+,
6%d&[῍ .\]"³´%῍ bc
- . / 6 % d &4 = , m . n o DT/
#$"O*h
10῎11῎ῌ )*+,
UUO p4=>14῎ῌ R
-./6%d&῍ .uvwxO
῍ AT1a ῌῒῑῒ῍ 4῍ )*+,-./B
"
µwx#$"O?&¶(῍
.CD23῍ UUO "4
R&·9
qY+,ῌ
῍ .\]"³´%
Yl'(29῍῍
!4῍ ¸¹wx"
rd)*+,-./6%d&
bc#$
῍ rst-4῍ AT1a ῌ῏ῑ῏῍ 4῍
Ep 2.uvpA
῍ AT1a ῌῒῑῒ῍ 4῍ Ep 2.uv
ῌ
wxpA῍ )*+,-./
k2y AT1a 6" AngII B.CD
2/z!pA῍ 9>ῌ ?
@A῍ AT1a ῌ῏ῑ῏῍ 4 Ep UUO
113
!º&Z°„»¤^
326
ῌ
Figure 4. Type I collagen immunostaining in AngiotensinIItype 1a receptor AT1a and AT1a . Type I
collagen immunostaining on day 6 after unilateral ureteral obstruction UUO in the UUO-kidney of AT1a
without administration of Eplerenone Ep a, the contralateral non-obstructed kidney of AT1a without administration of Ep b, the UUO-kidney of AT1a with administration of Ep c, the
contralateral non-obstructed kidney of AT1a with administration of Ep d, the UUO-kidney of
AT1a without administration of Ep e, the contralateral non-obstructed kidney of AT1a without
administration of Ep f, the UUO-kidney of AT1a with administration of Ep g, the contralateral
non-obstructed kidney of AT1a with administration of Ep h. i: The positive area of type I collagen
was assessed semiquantitatively as described under Materials and Methods. Values are the meanSE. ,
AT1a with UUO and without administration of Ep; , AT1a with UUO and administration
of Ep; , AT 1 a with UUO and without administration of Ep; , AT 1 a with UUO and
administration of Ep. p
0.05 compared to the AT1a with UUO and without administration of Ep.
Original magnifications, 200.
114
327
Figure 5. Type IV collagen immunostaining after unilateral ureteral obstruction UUO in AngiotensinIItype 1 a
receptor AT1a and AT1a . Type IV collagen immunostaining in the UUO-kidney of AT1a without administration of Eplerenone Ep on day 4 a, the UUO-kidney of AT1a without
administration of Ep on day 6 b, the contralateral non-obstructed kidney of AT 1 a without
administration of Ep on day 6 c, the UUO-kidney of AT1a with administration of Ep on day 4 d,
the UUO-kidney of AT1a with administration of Ep on day 6 e, the contralateral non-obstructed
kidney of AT1a with administration of Ep on day 6 f, the UUO-kidney of AT1a without
administration of Ep on day 4 g, the UUO-kidney of AT1a without administration of Ep on day 6
h, the contralateral non-obstructed kidney of AT1a without administration of Ep on day 6 i, the
UUO-kidney of AT1a with administration of Ep on day 4 j, the UUO-kidney of AT1a with
administration of Ep on day 6 k, the contralateral non-obstructed kidney of AT 1 a with
administration of Ep on day 6 l. m: The positive area of type IV collagen was assessed semiquantitatively
as described under Materials and Methods. Values are the meanSE. , AT1a with UUO and
without administration of Ep; , AT1a with UUO and administration of Ep; , AT1a with
UUO and without administration of Ep; , AT1a with UUO and administration of Ep. p
0.05
compared to the AT1a with UUO and without administration of Ep. p
0.05 compared to the AT1a
with UUO and without administration of Ep on day 4 after UUO. Original magnifications, 200.
115
328
ῌ
Figure 6. Histological evaluation of tubular dilatation in AngiotensinIItype 1a receptor ῎AT1a῏ ῌῐΐῐ῍ and AT1a
ῌ῔ΐ῔῍. Representative photomicrographs of periodic acid-Schi# stained sections on day 6 after unilateral
ureteral obstruction ῌUUO῍ from the UUO-kidney of AT1a ῌῐΐῐ῍ without administration of of Eplerenone
ῌEp῍ ῌa῍, the contralateral non-obstructed kidney of AT1a ῌῐΐῐ῍ without administration of Ep ῌb῍, the
UUO-kidney of AT1a ῌῐΐῐ῍ with administration of Ep ῌc῍, the contralateral non-obstructed kidney of
AT1a ῌῐΐῐ῍ with administration of Ep ῌd῍, the UUO-kidney of AT1a ῌ῔ΐ῔῍ without administration of Ep
ῌe῍, the contralateral non-obstructed kidney of AT1a ῌ῔ΐ῔῍ without administration of Ep ῌf῍, the UUO-kidney
of AT1a ῌ῔ΐ῔῍ with administration of Ep ῌg῍, the contralateral non-obstructed kidney of AT1a ῌ῔ΐ῔῍ with
administration of Ep ῌh῍. i: The degree of tubular dilatation was assessed semiquantitatively as described
under Materials and Methods. Values are the mean ῒ SE. , AT 1 a ῌῐ ΐ ῐ῍ with UUO and without
administration of Ep; , AT1a ῌῐΐῐ῍ with UUO and administration of Ep; , AT1a ῌ῔ΐ῔῍ with UUO and
without administration of Ep; , AT1a ῌ῔ΐ῔῍ with UUO and administration of Ep. Original magnifications,
ῑ200.
116
329
Figure 7. Histological evaluation of tubulointerstitial damage in AngiotensinIItype 1a receptor AT1a and
AT1a . Representative photomicrographs of Azan-Mallory stained sections on day 6 after unilateral
ureteral obstruction UUO from the UUO-kidney of AT1a without administration of Eplerenone
Ep a, the contralateral non-obstructed kidney of AT1a without administration of Ep b, the
UUO-kidney of AT1a with administration of Ep c, the contralateral non-obstructed kidney of
AT1a with administration of Ep d, the UUO-kidney of AT1a without administration of Ep
e, the contralateral non-obstructed kidney of AT1a without administration of Ep f, the UUO-kidney
of AT1a with administration of Ep g, the contralateral non-obstructed kidney of AT1a with
administration of Ep h. i: The degree of tubulointerstitial damage was assessed semiquantitatively as
described under Materials and Methods. Values are the meanSE. , AT1a with UUO and without
administration of Ep; , AT1a with UUO and administration of Ep; , AT1a with UUO and
without administration of Ep; , AT1a with UUO and administration of Ep. p
0.05 compared to
the AT1a with UUO and without administration of Ep. Original magnifications, 200.
117
TUVW
330
XYῌZ[\]
1
Figure 8. This study proposed a mechanism, by which Eplerenone ῌEp῍ attenuated the tubulointerstitial fibrosis in
unilateral ureteral obstruction ῌUUO῍. Increased plasma Angiotensin II ῌAng II῍ in UUO stimulates
aldosterone secretion through both Angiotensin II type 1a receptor ῐAT1aῑ and AT2 receptors in the adrenal
cortex. Ep prevents the binding of the aldosterone with its receptor, modulates the AT1a receptor-mediated
signal transduction pathways and ameliorates the tubulointerstitial fibrosis induced by UUO.
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Abstract
Eplerenone, an aldosterone antagonist, attenuates renal fibrosis
via modulating angiotensin II type 1a receptor
῔mediated signal transduction pathways
Hiroshi Miura1, Atsuko Ikemori-Kamijo1, 2, Ayako Obama1, Takeshi Sugaya1,
Takashi Yasuda1, and Kenjiro Kimura1
In addition to Angiotensin II, aldosterone is known to aggravate the progression of tubulointerstitial
damage. In this study, we tested the hypothesis that function of AT1a receptor that is the receptor of
angiotensin II ῌAng II῍ and an aldosterone is connected with progression of tubulointerstitial damage in a
model of severe renal interstitial injury. We used a unilateral ureteral obstruction model with Ang II type 1
a receptor ῌAT1a῍ knockout mice ῎AT1a ῌΐῒΐ῍, nῑ40῏ in which Ang II signaling via AT1a was blocked; wild
type mice ῎AT1a ῌῐῒῐ῍ , nῑ41῏ were used as controls. AT1a ῌΐῒΐ῍ῌnῑ20῍ and 21AT1a ῌῐῒῐ῍ ῌnῑ21῍ mice
were administered Ep by oral gavage at a dose of 200 mgῒkgῒday for seven days before unilateral ureteral
obstruction ῌUUO῍ until they were sacrificed on days 2, 4, and 6 after surgery. EP did not reduce blood
pressure in either AT1a ῌΐῒΐ῍ or AT1a ῌῐῒῐ῍ mice. In AT1a ῌῐῒῐ῍ mice with UUO, administration of Ep
inhibited the expressions of collagens type I and type IV and attenuated tubulointerstitial damage. These
results suggested that Ep suppressed tubulointerstitial fibrosis in the AT1a ῌῐῒῐ῍ ῌ In AT1a ῌΐῒΐ῍ mice with
UUO, and without administration of Ep, the expressions of MCP-1, and collagens type I and type IV were
significantly reduced, and the tubulointerstitial damage was milder than that in AT1a ῌῐῒῐ῍ mice without
administration of Ep. These results suggested that blocking the action of Ang II via AT1a inhibited the
development of tubulointerstitial damage. In AT1a ῌΐῒΐ῍ mice with UUO, administration of Ep did not
change the expressions of MCP-1, collagens type I and type IV, and the degree of the tubulointerstitial
damage. Blocking the actions of both Ang II via AT1a and aldosterone did not produce a synergistic e#ect
for preventing the progression of tubulointerstitial damage. In conclusion, Ep ameliorated the tubulointerstitial fibrosis induced by UUO via modulating the AT1a-mediated signal transduction pathways.
1 Division of Nephrology and Hypertension, Internal Medicine, St. Marianna University School of Medicine 2ΐ16ΐ1
Sugao, Miyamae-ku, Kawasaki-shi, 216ΐ8511, Japan
2 Department of Anatomy, St. Marianna University School of Medicine
121