12
臨腹膀腣膄腵膉膒臸膚臁臇 Vol. 34, pp. 415426, 2006 腜腚腒腟腇腝腧腁腃 2 腙腣腤腦腍腊腋腅腡腘腥腠腖腄腗腨腂腆 腞腔腈腋腐腈腄腢腂腆腓腞腉腎腌腑腑腀腏腅腛腕 腑腖 腓腙 腏腐腓腌 1 腏腊 腗腆 2 腑腆 腏腊 1 腉腔 腑腍 2 腕腘 腊腚 腙腆 1 : 18 9 20 膹膫膆腘腊腑腛臣臠臙腗臯膅膌臄腗膋腑腓腁腞腎腗臷腗臟膠臔腖臃膏腭腴膂腭腗膝膺腉腠腒腂腟膯膢腓腘腊腑腛臣臠臙腖腄腈腟臷腗臟膶腢臢腌腟腏腛腖膸膆腔臠 臚膖腗膕臬腢腪腆腔腊腒2 腊腑腛腽腳膁腾腱腴腖 b1 臫臹臵臋臻腓腁腟膎臃 腮膀腷膃膃腀膁 CEL腉腯腊腑腛臣臠臙腗臷腄腜腙臃膏腭腴膂腭腖臶腌腟膍膤腢膮 腊腏2 腊腑腛腽腳膁腓腁腟 Wistar fatty rat WFR臶臘腔腊腒 Wistar lean rat WLR0.7臞膎腗臜臞 NS腚腏腘 7膹臞膎臞 HS腢臧膳 6 臑腅腝 14 臑腚腓腊腏腉腝腖膹臞膎臞膦腖CEL 300 mgkgday 腢膩膴腉腯腊腏膗膦腖腄腂腒臏臒膟膫 SBP膫臦腤膄腭膀膄臼腑腁腣膁腶腺膄腕腻腵 UAE臃膏腭腴膂腭腗臅腓腁腟8-hydroxy-2-deoxyguanosine 8-OHdG腗臱腢腊腏腉腝腖臆膡臵腖腑腂腒膛臣臃 臮臗膣膷臮腓腁腟 mangan superoxide dismutase Mn SOD腗臺腖腗膲腢腥腦腭腰膄腶膃腱腴 腓膮腊腏SBP, UAE腄腜腙腑腁 8-OHdG WFR-HS 膦腖腄腂腒 WLR 膦腄腜腙 WFR-NS 膦腔膙腊腒 8 腅腝 14 腚腓膈腖臯膐腊腏CEL 腉腯WFR-HSCEL腓腘WFR-HS 腓臯膐腊腏 SBP 8-OHdG 膈腖臢腉腠UAE 臢膧膵腖 腁腐腏腚腏膫臦腤膄腭膀膄臼腘 CEL 腉腯膦腓膈腖臢腉腠腏腉腝腖 WLR 膦腓腊腒腂腏 Mn SOD 腗臺腖腗膲腘 WFR-HS 臢腊腏腆WFR-HSCEL 膦腓腘膔膧膵 腖腁腐腏膇臛腗臤自腅腝CEL 腘膸膆臀腖膐腃腊腑腛臣臠臙腗臟腖膝腡腟臃膏腭腴膂 腭腢臡臥腊臠臙腗臷腢膪膱腉腍腟膑臣腆臉膼腉腠腏ῐῌ῏ 膹膫膆2 腊腑腛臃膏腭腴膂腭8-hydroxy-2-deoxyguanosine, mangan superoxide dismutase 腊腑腛臣臠臙腘膟臠臭腗膰膌臊膜腗膋腑腓腁 腛腣腗臌膽腘臠臰腖臆膡臵腖臲膿腌腟 1腊腑腛腗臠臰腓腘腼腬膄腩腥腻膾腦腎腗腪膁 腫腀腭 腓腈 腆臛臖腊advanced glycation end- products AGE腸膀腧腀膁膩腗膛臣膏腷膃腲腤膄腨腵腀腯 C PKC腗膛臣膏腖腜腞膏腭腴膂腭腆臟腌腟 2-6臃膏腭腴膂腭腖腜腟 DNA 至臕腗臅腔腊腒 8-hydroxy-2-deoxygua- nosine 8-OHdG腗臂臧臯膐腆腃腪腉腠腒腂腟 78-OHdG 腘膘臃腆腕臃膏臐臝腢臍腈腏腔腇 腖臧腋膞腄腜腙腊腑腛臙腄腜腙臟膶臈腖臯膐 1 臨腹膀腣膄腵膉膒臸膚 膒膚臠臰膹膫膆2 臨腹膀腣膄腵膉膒臸膚 腫腴415 203

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Page 1: 2 ˘ˇˆ˙ ˝˛˚ ˜˚ ˆ˙!˝#$%%&’ ()igakukai.marianna-u.ac.jp/idaishi/www/344/21-34-04Tadahisa Tomoh… · UAE in the WFR-HS group was markedly increased in the other three groups

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�WLR� - 0.7�cW��dc �NS�eC� 7��cWc �HS�@fg 6hij 14he% kKC� 6j-�cWcl-� CEL @ 300mg�kg�day @mn��KC� ol-=98pqr�� �SBP�� �st�0��u� �!�Ovw�"#$ �UAE�� ./0120�x%%&:�! 8-hydroxy-2�-deoxyguanosine �8-OHdG� $�y&@KC� 6j-z{T-$98� |�.}~��}%&:mangan superoxide dismutase �Mn SOD���'(�@��0��vZQ1)%a�KC� SBP, UAE� =\]�! 8-OHdG$�� WFR-HSl-=98WLRl=\]WFR-NS lF*�K8 8 h+ij 14 h+e%,�- �KC� CEL ��l �WFR-HS�CEL�%�� WFR-HS% �KC SBPF 8-OHdG$4,�-�A67� UAE��A��-&�C� eC� �st�0��u� CEL ��l%,�-�A67C� 6j-WLR l%(�K89C Mn SOD ��'(�� WFR-HS %�AKC4� WFR-HS�CEL l%��-��-&�C� �����ij� CEL �E��.-���������*�-3�:./0120@��K� ����)@��6�:���4��67C�

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����������7�� Ha �� ������������������������������ �� ���� 8-OHdG ������ �����8�� !"#$�� �����%&�'()��������*�+�,"���� �-./�����9�10�� 0�"�����������1��2�3��� �4*5���6��� ��7����*8�9:�� ����������;<���� =���2�>���*2?� @�ABC��D� 12?�EF�G'H�:�11�� 2 I���%&�!"#�G'H�)����D� $7�*'H�(%����;<�&��J�'(�� �-�K���FF:�12���������)���G'H� �LM)�*+N,-�.��*/N,-�O0�� ��;DLM)1H�23�LM)P45P�Q"�� � ��LM)R$S�T67U�8Q*�2�*�� �LM)*/N,-�O0 V����WXY���Z�[9���D13�� 0�"��������G'H5��� �LM)*+N,-�:;�\<]=�^�"LM)1H�'(��>_]=���"��WXY��� �ACE� ?`a@AbBS��WXY��� �AT� IIcC)DdB��49:� ��.���Q"14�15�� �K�*�� ACE?`a@AbBS AT II cC)DdB� �LM)e�EF]=�:�"�PG�fH!"Gghij'��;D�T6�1�H���kI-:�15�� 0 9l�JK��fH]=�L'(>_]=�mF� bcC)nMB�NO�"� bcC)nMB�������5�B�o��h�pc��7U*q�P�IK:!Dr=����*KC"�� st� L'(�]=� ACE ?`a@AbB�Q���>_]=�mF ������"16�17�� uvr=�"w#xh�yyz�'(\<�� b1 :;�cC)nMB9:��� G'H�RC"��������&�.��4,�F��S�{|T�"��*��0 9U}~9� G'H��)��" 2I���������� b1 :;�cC)nMBw#xh�yyz��VW�"�Q�����&�.�q�;�*4,�.�K� ���&����AW��#$�� ��.�����)B��|T�"�

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�� ��b1 :;�cC)nMB9:�w#xh�yyz� �CEL� XUYB������Z�;D+��"�CEL [%\w]�^3��r=�"� U}~9� dmangan superoxide dismutase �Mn SOD�i�W�h�yz��d) �Santa Cruz Biotechenol-ogy, CA, USA�� 0��d a-tubulin �h�yz��d) �Abcom, Cambridge, UK� �r=�"��� ���Wistar fatty rat �WFR� � _`�a�� obese

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«¡�@����

- .

�� �/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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205

Page 4: 2 ˘ˇˆ˙ ˝˛˚ ˜˚ ˆ˙!˝#$%%&’ ()igakukai.marianna-u.ac.jp/idaishi/www/344/21-34-04Tadahisa Tomoh… · UAE in the WFR-HS group was markedly increased in the other three groups

�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.

^_`a bcde �418

206

Page 5: 2 ˘ˇˆ˙ ˝˛˚ ˜˚ ˆ˙!˝#$%%&’ ()igakukai.marianna-u.ac.jp/idaishi/www/344/21-34-04Tadahisa Tomoh… · UAE in the WFR-HS group was markedly increased in the other three groups

���������� ����� 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.

I$JKLM��N# bOPQRST�UV 419

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WFR-HS �������� WFR-HS�CEL ������ ��������� �Fig. 6�

� �

����� ����������� �!"��#$%&�'()*�+,-./&0� b1 12

/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.

ghij klmP n420

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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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����������������� ��� �������� ���������� ������������� � WFR-HS� �� !"##$%���&���������' �()*� �Fig. 1�� Miura ���+ ���� ���,��-./ �� !"##$%� 60123��&��� ��� �&���4��5��� ������' ����� �6�4b1 7� 8!6���9� b2 7� :; ��"<=>#2:;6��?@$�AB ��9��CD�E��25�� ���5� WFR-HS�CEL��F�6�9G�� b17� 8!HIJ�@� b2

7� :;�"<=>#2:; ��%&�K�L'6� ��(M����(6�����E��CD�E�25�� G�N)�����OP Q�� �� !"##$%�F�RB �D����(��L&�6� S��+��Q�26�� T��ABJ4� �� !"##$%���� �*U%VWX+�������L&�YZ�������� �������� �9�L&�[���\)E�� Malminiemi �4�� !"##$%�� ���OPG�4]^_!.`a��OPJb%c$d,efgh���-Q�i�.jXd!X-��(�� /���k0�l�E����+���27�� G�� �� !"##$%4���OP)� ]^_!.`a�OP Q�jXd!X1m�k0&����+)E��28�29�� ����J4n2�����34�]oXpqrs5ts6�K 7@�u8�v��� ��� ��9�����30�� wx�ABJ4� �.jXd!X-4� WFR-NS �Q�i WFR-HS �JWLR ��9y��' ������ �Fig. 3���� !"##$%�� �WFR-HS�CEL� �J4� ��.jXd!X-��' �(��� z:� b%c$d,efgh���-4 WFR-NS�� WFR-HS �Q�i WFR-HS�CEL �� 3�1J{|;}YZ���� �� !"##$%�� ����-��(4l�E��5�� ���5��� !"##$%4� ����~<��������4 �.�jXd!X- ����(6�4�����-��()*�RB4=��CD�E�� wx�.jXd!X-�(�� �������|���4>?J��� �� !"##$%��.jXd!X-��(�9��&�@ S

��A&�BC����)� ���� QI�������~<�DE ��d/�d�FC�G��B������?�� �5�v�9�10�� ��d/�d ���HIJK����DNAL���9�H�7I��9�5�v�31�32�� ��J��H�7I��� �" DNA� s5�"�`����� W/cX�!U DNA ���� *J9W/cX�!U DNA4�� �H�7It&@� W/cX�!U"J��J!X�� �� ATP ~M ��� d$�$��oj� �O2�� �����N�OP)E���8-OHdG 4��N �������J�H�7I�h � bU�X�� 8��QN�R��)E��SJ��� 8-OHdG �TM4�t�����t�4�D�hJ��&���U�E��7�8���V� Hinokio �4�K~<��&����OPJ4�* 8-OHdG +�W���������+�� �* 8-OHdG � DNA �� ���$�$���[���)E�33�� G� Kanauchi �4�����XY�J��sK����* 8-OHdG ���Z&������ ��d/�d�������4�[ �����CD�E�34�� wxWFR-HS�J�* 8-OHdG��WLR�tWFR-NS ��9y� 80\���'������� �Fig. 5�� �AB������]5�����J4����d/�d�^D����CD�E�� Saez �4� ���OP b 7� 8!_J��U¡¢#$%G�4£¤"##$%�¥`¦ � �*� 8-OHdG �t§����d/�d�$�$�ab�� F�RB��9 ��d/�d�L'�l�E�����35�� ¨�c 9 b7� 8!_���d/�d �&���RB S�@S��+���35�36�� T��ABJ4� 2 ©���ª«%-./ Q��� !"##$%��* 8-OHdG���' ��������� �Fig.5�� wx�U¬4� EGJ?�� )E���5��� !"##$% ����d/�d���RB�d� ¬e��� ��� ��d/�d�^D4­® W/cX�!U" QI���Nf¯°±N������9²E�� T "���Nf¯°±N4 SOD� �^-$³� b%^��X´%��µ¶$³���g·�� s5" QI���¸¹�º8�hn���� *J9 Mn SOD

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�����������35�� ����������� Mn SOD ����������� ��� !"�#"$%�&'()*�+����,��-./����35�� 0��1�2�� 34�WLR� 5�67���� Mn SOD �8967$�WFR-HS ��:/�� ; <)=>>?@ABC�,��:�DE/��,�2�� ; <)=>>?@A$ ���F���GHIJ��A����K�� !"�#"�LMN�O��P�,��Q�R-�� �Fig. 6�� �2� !"�#"$� SJ TUV)?�WX@����YZ�� ��[SJ T\]�^_`Aa��bc����d� ef� g !hT[i�jkSJ T���l�$m !YZn�o�Ap��q�RrsN�t��P����uv�wxB�� ��$GHIJ���Iy�Kz/{� |�Kz}~�g�A !"�#"��K�������,����2A[C�� l� ���F���GHIJ��A���; <)=>>?@�� �[��o��-�� !"�#"��K��:�Iy���N���J�-./���ef�A���GHIJ�����F����U� �������c������

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����AP��� ���A��������)*'������������ g�¡� l�¢£���¤����)*'�����I¥�¦ ��§����¨©�ª���«¬� ��)*'�������¥­®¯°±²��³´�µ�A¶R��·�v¸lN�

����

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�������� b�� ������ 425

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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

���� ���� 426

214