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� � ����������Vol. 34, pp. 415�426, 2006
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Zucker rat�Wistar Kyoto rat��b��"���9� _`�Ac�" 2I������������1981d�6���"18�� WFR �fa�fa� [� 6�e;D_`�a�� !"G'�� Go��h�'��o��h�fd�*q� 2I����������� l�� Wistar lean rat �WLR� �fa�-��ghBi�������8��;DjW�c�k�h�����8�8��lm��,n}~�o19Zp� m����� U}~9[� 6�eK 14�e!9q]¡� 0.7� \w�>¢rs\ �CE2 £¤¥t� CLEA� u��¦ �WFR�NS ¦� n�7� � 7�\w�>¢G\wv§¦ �WFR�HS ¦� n�7����WFR�HS¦�w#xh�yyz� 300mg�kg�day �VW�"¦ �WFR�HS�CEL ¦� n�7��]��"� U}~�w������ WFR�HS�w#xh�yyz�� 30, 100, 300mg�kg�day �0�¨�©ªVW�� fH4,�«�"=��300 mg�kg�day� �U���=�"� ��.x¦��� WLR �rs\v§¦ �WLR�NS ¦� n�7�� G\wv§¦ �WLR�HS ¦� n�7� �;<WLR�HS ¦� CEL300mg�kg�day �©ªVW�"¦ �WLR�HS�CEL¦� n�7��]��¬ 6¦�F��|T�"� J��,n� k�h�����8�8��lm��,n}~�o19® �231�C�� ¯ �555��� y�x� �6 °18 °�� ±z²]U9¥³�"� U}~U���,n´µ��;�{¶� | �|}· 0510021� �c���D� �,n�A��XUB%��¸~� ��¹��JC"��� ������º»°�[� 6�e��� 8�e� 10�e�;< 14�e� tail-cu# ��=���,-'H��
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���� ���������� 5����������������� ��������� ����� 6 ��� 8 ��� 10 ����� 14������������������ ����� !� �"!�#!$� %&�� BeckmanCoulter, Fullerton, CA, USA�� ��'()!����*+,��'()!-+,� "./0� ���� � 8-OHdG �New 8-OHdG ELISA kit, �123'� 45� ������ 6� 14 ���7!,.'89�'��:�;��<=>?��� =����@AB�BC���� !� �"!�#!$� %&�� Beckman Coulter, Fullerton, CA,USA� �BC3!DE!� ��8D3!DE!ELISA -+,� "./0� ��� �������� �������� �Intraperitoneal glucosetolerance test� ��������� � 14��*+,��FG�H��I'J�D�KLF��M�� 8HNOP��B������� 50� I'J�D 2 g�kg ��;����� 30 � 60 � 90 ��� 120 ��B�������� B�����QRSB���&��I'TD,U�D� VWXYZ�[\1]^� �_`�������� ���� !�"#��$%&'(14��*+,a�� RPMI 1640 !b �Sig-ma-Aldrich, St Louis, MI, USA� >"#�$c��� 80, 150��� 200mmd+"e����"�8!I%��@a&fghij'�()��� ()��ij'Q*+,c-b �50 mM Tris HCl, 1mMEGTA, 0.001�#37k�!� pH7.40�>lm�n3o�� 15,000�g� 4�C> 10 Npq�.hr�sC��t��� ,c��*+Q� Bio-Rad pro-tein assay kit �Bio-Rad Laboratories, Hercules, CA,USA� ����*+������� �rur*+�� 20mg �/v�� SDS w!k'x0b�yz�� 100�C > 2 N{1��2|}��L3���� L3Q 10� SDS ~E��E'�)��'��k*3�� 2|}����� 2|}����'�Hybond-ECL �Amersham biosciense, Buckigham-shire, UK� �4��� 5� D-�)'�>� 4�C��5(#+-!Ix0��M�� �� 0.1� �v�v�Tween 20 �6 20mM Tris ��b �TTBS, pH7.5�> 200 7������ Mn superoxide dismutase
�SOD� �w0�'������> 2 HN8������ TTBS >����M�� �h�� 5000 7����l�D*��+"e7'�-"����HRP� 9���w0�:�Q���D IgG �'�' �ICN Pharmaceuticals Inc., Aurora, OH, USA�> 1HN8���� ECL plus �Amarsham� ����:���� Fuji Film Las-3000 �Fuji film, Tokyo���������� )*+,� ;��% Stat View �ver. 5.0������ .hr���Q���9� ¡>¢��� <�N=£QANOVA > ¤�¥>�� Fisher %�����¦M�� ¤���¦�§zQ� Mann-Whitney U¨������ ©ª? 5� @A�6«¡�@����
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�� �/0 1/2345��6789:���;<�F?¬H�6���'QWLR �> 193.00�7.35 g, WFR �> 227�3.25 g >�M�� �FG�H�14 ���'Q ��®6«�¯°����WLR-NS �� WLR-HS ���� WLR-HSCEL � 3�N¦h�� WFR-NS �� WFR-HS���� WFR-HSCEL � 3 �N>Q'�6«¡Q>±¦gM� �Table 1�� a�Q WFR
�> WLR � 3 ��=£�6«�¯°����WFR-NS �� WFR-HS ���� WFR-HSCEL �>6«¦¡Q>±¦gM� �Table 1��14 ���BC���� !������� BC���� !�Q 6�>6«¦¡�>±¦gM��Table 1���� =>?�@�;<6 ��>Q �>6«¦¡�¢�¦gM��WFR-HS � �173.44�1.04mmHg� Q 8���@s²�� 14 ��>Q 204.00�0.87 mmHg >�M��Fig. 1�� WFR-HSCEL �® 8� �153.42�0.73mmHg� �@s²�� 14��>Q 178�1.00mmHg>�M��� WFR-HS ��=£�6«�B³�r���� �C 4�>® 6���=£� 14��>Q� ´D�s²���� �Fig. 1���� ��������345���������;<14 ����M�I'J�D�KLF���Fig. 2�¢µ� I'J�D�K¶�µ>�WFR�
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�WLR ���� 1.2����� ���� ������� 30���� WFR �� WLR ��������� �� WLR-NS �� WLR-HS � �! WLR-HS�CEL �" 3 �#$�!�WFR-NS �� WFR-HS � �! WFR-HS�CEL�" 3�#�%&'�()* �Fig. 2�� ���+,-./ CEL ����012�3�45��6���789�:9�� ;<=+� �>�?@
�� WFR-NS�� WFR-HS� �!WFR-HS�CEL ��� WLR �5AB %&�CD � �Fig.2�� > WFR-HS�CEL � �1.32�0.23 ng�ml��� WFR-NS � �2.71�0.36 ng�ml�� WFR-HS ��2.82�0.30 ng�ml� E�� ��789�:9�%&�FG � �Fig. 3���� ��������� �HIJ"KL5 �MNO�PQ9"RST�UVW�@X � Y"Z[� Fig. 4 �� �� 6\]
Table 1. Characteristics of Wistar Lean Rat �WLR� and Wistar Fatty Rat �WFR� with Normal or High SodiumLoad at 14 Weeks of Age
Fig. 1. E#ect of celiprolol on systolic blood pressure �SBP� during the experimental period.SBP in the WFR-HS group was significantly higher than that in the other three groups at the ages of 8 to 14
weeks. SBP in the WFR-HS�CEL was significantly inhibited, compared with WFR-HS. The data are repre-sentative of seven independent experiments. Details are described in Materials and Methods. �p�0.01 vs.WLR-NS, WLR-HS, WLR-HS�CEL and WFR-NS, �p�0.01 vs. WFR-HS, Wistar lean rats; WFR, Wistarfatty rats; WLR-NS, WLR fed a normal-salt diet; WLR-HS, WLR fed a high-salt diet; WLR-HS�CEL,WLR-HS treated with celiprolol; WFR-NS, WFR fed a normal-salt diet; WFR-HS, WFR fed a high-salt diet;
WLR-HS�CEL, WLR-HS treated with a celiprolol.
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206
���������� ����� 14 ���� WLR-NS � �0.38�0.02mg�day�� WLR-HS� �1.24�0.01mg�day� ��� WLR-HS�CEL ��1.68�0.01 mg�day� � 3 ������������ ��� WFR-NS � �14.40�1.2mg�day���� WFR-HS � �69.55� 4.36 mg � day� ��WLR ������������ WFR-HS ���WFR-NS� 5!��������WFR-HS�CEL � �56.50�6.64mg�day� � WFR-HS ����"#�$%&'()*+,-�./01� ���Fig. 4��
�� �� 8-OHdG ����234564�7898��:$% -OHdG-�;<��� 6������������ ����=� WFR �� 8���>��� 14���� WFR-HS � �2.45�0.55mg�ml� ��� WFR-HS�CEL � �1.53�0.12mg�ml� ��?� 4�������@A�:B�� ��� WFR-HS�CEL�� WFR-HS �������CD�:B��Fig. 5���� ��� �� Mn SOD �����WLR ��EF�:B� Mn SOD �GHEF�
Fig. 2. Results of the intraperitoneal glucose tolerance test �IPGTT� in WLR and WFR fed a normal-salt diet,high-salt diet or high-salt diet treated with celiprolol at 14 weeks of age.
��p�0.05 and �p�0.01 vs. WLR-NS, WLR-HS, WLR-HS�CEL. The data are representative of fourindependent experiments. Details are described in Materials and Methods. The abbreviations are the same as
in the legend to Fig. 1.
Fig. 3. E#ects of celiprolol on plasma insulin levels after overnight fasting in WLR and WFR fed a normal-salt diet
or high-salt diet at 14 weeks of age.
�p�0.01 vs. WLR-NS, WLR-HS and WLR-HS�CEL, �p�0.01 vs. WFR-NS and WFR-HS. The data arerepresentative of six independent experiments. Details are described in Materials and Methods. The
abbreviations are the same as in the legend to Fig. 1.
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WFR-HS �������� WFR-HS�CEL ������ ��������� �Fig. 6�
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/345678���9:;<=>>?@�AB�CDEFG0�� �������� �� HI/JK-LMN�OKP�QR�!S�ETU� VWKP�XC�#$%���11�� 2 Y���Z[�#$%�\]-^_&�`a� b�����c�#$%�d���&�ef&OU� ������
Fig. 4. E#ects of celiprolol on urinary albumin excretion �UAE� during the experimental period.UAE in the WFR-HS group was markedly increased in the other three groups. The data are representative
of 10 independent experiments. �p�0.01 vs. WLR-NS, WLR-HS and WLR-HS�CEL, ��p�0.05 and �p�0.01vs. WLR-NS, WLR-HS, WLR-HS�CEL and WFR-NS, �p�0.01 vs. WFR-HS. Details are described inMaterials and Methods. The abbreviations are the same as in the legend to Fig. 1.
Fig. 5. E#ects of celiprolol on 8-hydroxy-2�-guanosine levels in the urine during the experimental period.The 8-OHdG level in the WFR-HS group was markedly increased in the other three groups. The data
are representative of six independent experiments. �p�0.01 vs. WLR-NS, WLR-HS, WLR-HS�CEL andWFR-NS, �p�0.05 vs. WLR-NS, WLR-HS and WLR-HS�CEL, ��p�0.05 and �p�0.01 vs. WFR-HS, �p�0.01 vs. WFR-NS. Details are described in Materials and Methods. The abbreviations are the same as in the
legend to Fig. 1.
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������������ �� �������������� ������������ �������� !"# $�%&'� (�)*+� ,�-.�/19�20�� ��0��%1� ����*23450����1� 67����#�8�9:��;'� 67����<���=�#�><���?@��ABC 67�����D�E""# �FG�-.�/21�� "�HI�� ��<���JHKL�?@�M� �NO��PQR�SRTUVRWRX�Y0Z� [R\]R^���M*_ �8�GG2��/"# �FG�-.���22��".`%� �������� ��0�������a��bcdefg1&`'h�-.�;Fi� -F������`�1jk����l����mn��*/� o� pq��/� Wistarfatty rat �WFR� 1!"�#�� �� � �[R\]R��� [R\]R$r0*_�0s�nt 2u ��bcdefg%� �vw�%x"#�y'�����#�� ��0���67��&�'(%&67�z)*�!{;y|}~RT��+��=� ,�F.18�� `���F1� ��� ��0�����%&��Sd��R-./�j��� (�����0��� Tumor growthfactor-b �1�j�� Mitogen-activated proteinkinase �Y0Z 92"#��FG���23��("%� WFR ��vwv%x��3�� ��4�52��������3)�� ������3)�� -F�����0���3���t/�&2��l���� �6� ��0�����
��+4#��67�������#�7�^���� ACE &�����4� AT II�8�9r4 �:-.���14�15�� �G�* F� ACE &�����4� AT II �8�9r41� �67���;8^��y�<���`�1��=�jk*_�>?n&15�� ("%o�1450���^�#���7�^���t b �8��@4�AB��� b�8��@41� ��0���+4�[R\]R �0��M� ¡sC¢�kZ*_�>?D &'� EF52�+4#��1&`'�/F.*G��� �G�����^�� ACE &�����4#GH��7�^���t"# �m-.�16�17�� Rudberg F1� b1�8��@4%&}g£¤¤¥d1� ACE &�����4¦§e£]d#I¨���©�NOª�*«DGF�¬��J< ®-.�"#��m��/16��w¯°]£¤¤¥d1 b2 �8�±²�y��=�^��K��t b1�:��8��@4%&'��6� w¯°]£¤¤¥d ����efg�;/���^��³G��©�LM¬��N´"# �m-.�24�� �G�w¯°]£¤¤¥d�y����µK�� ��0������3)��t/�� ¶��l����m1*/� WFR ��vw%x �WFR-HS� ��?@,��1� 3·�%& WLR �#I¨��� �� 8¸Oy'¹º��14¸O`%P��¹º�� �Fig. 1�� `���Sd��R-./n»�j���/� �Fig. 4�� 3·�� WLR �3��vw�%x��n��¹º���Sd��R-./�j��,�*/"#� `�WFR �QCv%x��;/�n��Sd��R-
Fig. 6. E#ect of celiprolol on Mn superoxide dismutase �SOD� in glomeruli of WLR and WFR at 14 weeks of age.Fig. 6. shows the representative bands of Mn SOD protein expression analyzed using Western blotting.
a-tubulin was used as the loading control. The expression of the WFR-HS was significantly increased
compared with in the other four groups. The expression of the WFR-HS�CEL was significantly inhibitedcompared with in the WFR-HS. Details are described in Materials and Methods. The abbreviations are the
same as in the legend to Fig. 1.
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1� Andersen AR, Christiansen JS, Andersen JK,Kreiner S and Deckert T. Diabetic nephropa-
thy in Type 1 �insulin-dependent� diabetes: anepidemiological study. Diabetologia 1983; 25:
496�501.2� Brownlee M, Cerami A and Vlassara H. Ad-vanced glycosylation end products in tissue and
the biochemical basis of diabetic complica-
tions. N Engl J Med 1988; 19: 1315�1321.3� Greene DA, Lattimer SA and Sima AA. Sorbi-tol, phosphoinositides, and sodium-potassium-
ATPase in the pathogenesis of diabetic compli-
cations. N Engl J Med 1987; 5: 599�606.4� Koya D and King GL. Protein kinase C activa-tion and the development of diabetic complica-
tions. Diabetes 1998; 47: 859�866.5� Thuraisingham RC, Nott CA, Dodd SM andYaqoob MM. Increased nitrotyrosine staining
in kidneys from patients with diabetic nephro-
pathy. Kidney Int 2000; 57: 1968�1972.6� Kitada M, Koya D, Sugimoto T, Isono M,Araki S, Kashiwagi A and Haneda M. Translo-
cation of glomerular p47phox and p67phox by
protein kinase C-beta activation is required for
oxidative stress in diabetic nephropathy. Dia-
betes 2003; 52: 2603�2614.7� Leinonen J, Lehtimaki T, Toyokuni S, OkadaK, Tanaka T, Hiai H, Ochi H, Laippala P,
Rantalaiho V, Wirta O, Pasternack A and
Alho H. New biomarker evidence of oxidative
DNA damage in patients with non-insulin-
dependent diabetes mellitus. FEBS Lett 1997;
3: 150�152.8� Ha H, Kim C, Son Y, Chung MH and KimKH. DNA damage in the kidneys of diabetic
rats exhibiting microalbuminuria. Free Radic
Biol Med 1994; 16: 271�274.9� Kashihara N, Watanabe Y, Makino H, Wall-ner EI and Kanwar YS. Selective decreased de
novo synthesis of glomerular proteoglycans un-
der the influence of reactive oxygen species.
Proc Natl Acad Sci U S A 1992; 15: 6309�6313.10� Sugiyama H, Kashihara N, Makino H, Ya-
masaki Y and Ota Z. Reactive oxygen species
induce apoptosis in cultured human mesangial
cells. J Am Soc Nephrol 1996; 7: 2357�2363.11� Wenzel RR. Renal protection in hypertensivepatients: selection of antihypertensive therapy.
Drugs 2005; 65 Suppl 2: 29�39.12� Zatz R, Dunn BR, Meyer TW, Anderson S,Rennke HG and Brenner BM. Prevention of
diabetic glomerulopathy by pharmacological
amelioration of glomerular capillary hyperten-
sion. J Clin Invest 1986; 77: 1925�1930.13� Adler AI, Stevens RJ, Manley SE, Bilous RW,
GHIJ��A��� b¹�º»¼��O� 423
211
Cull CA and Holman RR; UKPDS GROUP.
Development and progression of nephropathy
in type 2 diabetes: the United Kingdom Pro-
spective Diabetes Study �UKPDS 64�. KidneyInt 2003; 63: 225�232.
14� Bjorck S, Mulec H, Johnsen SA, Norden Gand Aurell M. Renal protective e#ect of enala-
pril in diabetic nephropathy. BMJ 1992; 304:
339�343.15� Parving HH, Brenner BM, Cooper ME, deZeeuw D, Keane WF, Mitch WE, Remuzzi G,
Snapinn SM, Zhang Z and Shahinfar S. E#ect
of losartan on renal and cardiovascular compli-
cations of patients with type 2 diabetes and
nephropathy. Ugeskr Laeger 2001; 163: 5514�5519.
16� Rudberg S, Osterby R, Bangstad HJ,
Dahlquist G and Persson B. E#ect of angio-
tensin converting enzyme inhibitor or beta
blocker on glomerular structural changes in
young microalbuminuric patients with Type I
�insulin-dependent� diabetes mellitus. Diabe-tologia 1999; 42: 589�595.
17� Abbott KC, Trespalacios FC, Agodoa LY,Taylor AJ and Bakris GL. Beta-blocker use in
long-term dialysis patients: association with
hospitalized heart failure and mortality. Arch
Intern Med 2004; 164: 2465�2471.18� Ikeda H, Shino A, Matsuo T, Iwatsuka H andSuzuoki Z. New genetically obese-hypergly-
cemic rat �Wistar fatty�. Diabetes 1981; 30:1045�1050.
19� Parving HH, Andersen AR, Smidt UM, Hom-mel E, Mathiesen ER and Svendsen PA. E#ect
of antihypertensive treatment on kidney func-
tion in diabetic nephropathy. Br Med J �ClinRes Ed�. 1987; 294: 1443�1447.
20� Mogensen CE, Keane WF, Bennett PH, Je-rums G, Parving HH, Passa P, Ste#es MW,
Striker GE and Viberti GC. Prevention of dia-
betic renal disease with special reference to mi-
croalbuminuria. Lancet 199; 346: 1080�1084.21� Hostetter TH, Rennke HG and Brenner BM.The case for intrarenal hypertension in the ini-
tiation and progression of diabetic and other
glomerulopathies. Am J Med 1982; 72: 375�380.
22� Wenzel RR. Renal protection in hypertensivepatients: selection of antihypertensive therapy.
Drugs 2005; 65 Suppl 2: 29�39.23� Imai G, Satoh T, Kumai T, Murao M,
Tsuchida H, Shima Y, Ogimoto G, Fujino T,
Kobayashi S and Kimura K. Hypertension ac-
celerates diabetic nephropathy in Wistar fatty
rats, a model of type 2 diabetes mellitus, via
mitogen-activated protein kinase cascades and
transforming growth factor-beta1. Hypertens
Res 2003; 26: 339�347.24� Kakoki M, Hirata Y, Hayakawa H, Nishi-
matsu H, Suzuki Y, Nagata D, Suzuki E,
Kikuchi K, Nagano T and Omata M. E#ects of
vasodilatory beta-adrenoceptor antagonists on
endothelium-derived nitric oxide release in rat
kidney. Hypertension 1999; 33: 467�471.25� Miura A, Kimura Y, Inoue K, Matsuzaki T,Ochi S, Hamada K, Hayashi S, Tamura M,
Kano S and Kimura K. Pharmacological stud-
ies of celiprolol: I. Beta-blocking e#ect, intrin-
sic sympathomimetic activity, vasodilating and
hypotensive e#ects Nippon Yakurigaku Zasshi
1990; 5: 191�200.26� ����� ���� ��� �� ���������� celiprolol �NBP-582� ��������� . !"#$ 1990; 6 Suppl. 1: 95�102.
27� Malminiemi K, Lahtela J, Malminiemi O, Ala-Kaila K and Huupponen R. Insulin sensitivity
in a long-term crossover trial with celiprolol
and other antihypertensive agents. J Cardio-
vasc Pharmacol 1998; 31: 140�145.28� Brownlee M. The pathobiology of diabeticcomplications: a unifying mechanism. Diabetes
200; 54: 1615�1625.29� Dandona P, Thusu K, Cook S, Snyder B,Makowski J, Armstrong D and Nicotera T.
Oxidative damage to DNA in diabetes mellitus.
Lancet 1996; 347: 444�445.30� Malminiemi K, Lahtela JT and Huupponen R.E#ects of celiprolol on insulin sensitivity and
%&'( )*+, -424
212
glucose tolerance in dyslipidemic hypertension.
Int J Clin Pharmacol Ther 1995; 33: 156�163.31� Oksa A, Fedelesova V, Spustova V and DzurikR. Celiprolol improves glucose metabolism in
essential hypertension. Vnitr Lek 1998; 44:
63�67.32� Fung H, Kow YW, Van Houten B and Moss-
man BT. Patterns of 8-hydroxydeoxyguanosine
formation in DNA and indications of oxidative
stress in rat and human pleural mesothelial
cells after exposure to crocidolite asbestos. Car-
cinogenesis 1997; 18: 825�832.33� Hinokio Y, Suzuki S, Hirai M, Chiba M, HiraiA and Toyota T. Oxidative DNA damage in
diabetes mellitus: its association with diabetic
complications. Diabetologia 1999; 42: 995�998.34� Kanauchi M, Nishioka H and Hashimoto T.Oxidative DNA damage and tubulointerstitial
injury in diabetic nephropathy. Nephron 2002;
91: 327�329.
35� Saez GT, Tormos C, Giner V, Chaves J, Lo-zano JV, Iradi A and Redon J. Factors related
to the impact of antihypertensive treatment in
antioxidant activities and oxidative stress by-
products in human hypertension. Am J Hyper-
tens 2004; 17 :809�816.36� Castro P, Vukasovic JL, Chiong M, Diaz-Araya G, Alcaino H, Copaja M, Valenzuela R,
Greig D, Perez O, Corbalan R and Lavandero
S. E#ects of carvedilol on oxidative stress and
chronotropic response to exercise in patients
with chronic heart failure. Eur J Heart Fail
2005; 7 :1033�1039.37� de Cavanagh EM, Toblli JE, Ferder L, Pi-otrkowski B, Stella I and Inserra F. Renal
mitochondrial dysfunction in spontaneously
hypertensive rats is attenuated by losartan but
not by amlodipine. Am J Physiol Regul Integr
Comp Physiol 2006; 290: 1616�1625.
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Abstract
Celiprolol Attenuates Urinary 8-hydroxy-2�-deoxyguanosinein Type II Diabetes Mellitus Model Rats with
Diabetic Nephropathy and Hypertension
Tadahisa Tomohiro1, Yuko Takeba2, Takeo Satoh1,
Toshio Kumai2, and Kenjiro Kimura1
Oxidative stress may contribute to the pathogenesis of diabetic nephropathy �DN�, although thedetailed mechanism of reactive oxygen species �ROS� regulation is still unclear. This study examined thee#ects of celiprolol �CEL� as b1 selective adrenoreceptor antagonist, on the expression of ROS and
antioxidant on the renal function and on the relationship of these factors as well in the experimentally
produced diabetic model rats. Wistar fatty rats �WFR� as a type II diabetes mellitus model and Wistar leanrats �WLR� as a control were fed a normal-salt diet �NS� and high-salt diet �HS� from the age of 6 to 14weeks. Furthermore, WLR-HS and WFR-HS were treated by CEL �300mg�kg�day� simultaneously withHS. We then examined the blood pressure, urinary albumin excretion �UAE�, and urinary 8-hydroxy-2�-deoxyguanosine �8-OHdG� levels. The expression of antioxidant enzymes, mangan superoxide dismutase�Mn SOD� was analyzed in the glomeruli of the rats using Western blotting. By 14 weeks of age, theWFR-HS group exhibited hypertension and markedly increased UAE. The level of urinary 8-OHdG, a
marker of oxidative damage, in the WFR-HS group was also higher than that in the WLR-NS or WFR-HS
group. But in the WFR-HS�CEL group, blood pressure and urinary 8-OHdG were significantly loweredand UAE tended to decrease. The expression of Mn SOD proteins was significantly decreased in isolated
glomeruli from the WFR-HS group. But in the WFR-HS�CEL group, the expression of Mn SOD wasincreased compared with in the WFR-HS group. High expression of ROS and decrease in antioxidants were
seen in the glomeruli of diabetic rats with hypertension, suggesting that oxidative stress may be involved in
the development of DN. Furthemore, CEL lowered the blood pressure and reduces the oxidative stress
which contribute to the development of the pathogenesis in DN. CEL can play a more important role in the
treatment of DN with hypertension.
1 Division of Nephrology and Hypertension, Department of Internal Medicine2 Department of Pharmacology
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