#$ 164,7+61

Transcription

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7 $ Relaxation (%) A 5 1 +E(n=8) -E(n=6) Relaxation (% ) B 5 1 +E(n=8) -E(n=6) Relaxation (%) C 5 1 Log [ACh] (M) +E(n=6) -E(n=4) Relaxation (%) D 5 1 Log [SNP] (M) control (n=17) L-NAME (n=8) ODQ(n=4) Log [Sildenafil] (M) Log [ACh] (M) Relaxation (% ) E 5 1 control (n=8) L-NAME (n=8) ODQ (n=3) L-NAME + Apamin + ChTx(n=8) Log [ACh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

8 $ @=OA>HOI)IIDME.ECI ) =@6=>AJDAHA=N=JABBE?=?OB E@K?A@HA=N=JEMAHA>IAHLA@>AJMAA#=@'@=O,)I>KJ)+D Relaxation (% ) A (n=16) 19(n=17) 21(n=12) B Relaxation (% ) (n=8) 21(n=9) Relaxation (% ) Log [ACh] (M) C (n=13) 19(n=14) 21(n=12) 19+ODQ (n=4) D Relaxation (% ) Log [ACh] (M) (n=3) 19(n=9) 21(n=8) 19+ODQ (n=4) Relaxation (% ) Log [SNP] (M) E 15(n=8) 19(n=6) 21(n=4) Log [8-BrcGMP] (M) F % ofk + -contraction Log [BAY ] (M) 1 15(n=4) 19(n=4) 21(n=4) Log [L-NAME] %,=@ JDA?/2==C&H?/2-=@JDA?JH=?JEIE@K?A@>OJDAIOJD=IAEDE>EJH )-. ' #+ 6DAABBA?JIB JDAIK>ACK=O=JA?O?=IAEDE>EJH,352=@);" % E@K?A@ HA=N=JEI=HA=IIDMOEJDA'@=O,).HHA=N=JEANFAHEAJI,)IMAHA?JH=?JA@MEJD $ #)+,-HHAFEAFDHEA2 #BH@EBBAHA?A E- =N HE=CELA??AJH=JEBH#=@'@=O,)

9 $! 6=>A % 15-day-old 19-day-old 21-day-old E max pd 2 n E max pd 2 n E max pd 2 n ACh 51.84(14.8) 5.86(.42) (17.3) 6.17(.32) (14.7) 6.57(.32) 12 SNP 73.44(19.4) 5.47(.52) (22.9) 5.92 (.4) (12.4) 5.42 (.39) 12 8-Brc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

10 64 detachment of endothelial cells, leaving large areas of exposed subendothelial tissue (Fig. 3C). These irregularities of the endothelial surface were markedly less A B 19 d pulmonary side C 19 d aorticside D 21 d pulmonary side 21 d aorticside E 19 d endothelium denuded Fig. 3. Scanning electronic micrographs of the endothelial surface of DA rings from 19-day (A and B) and 21-day (C and D) chicken embryos. In the 19-day DA, the endothelium showed a smooth and continuous surface with small cytoplasmic structures (similar to microvilli) protruding toward the lumen. By contrast, the intimal surface of segments harvested from the pulmonary side of the 21-d DA (C), demonstrated an irregular endothelial lining with protrusion and detachment of endothelial cells, leaving large areas of exposed subendothelial tissue. These irregularities of the endothelial surface were markedly less apparent in the aortic side of the 21-day DA (D) in which only small areas of exposed subendothelial tissue were observed between endothelial cells. E: Exposed subendothelial fibers and absence of endothelial cells in a 19-day chicken DA segment after mechanical removal of the endothelium by gently rubbing the intimal surface with a metal rod. Bars 2 µm.

11 $# A - ACh + ACh 19 d + L-NAME 19 d control 21 d control.ec")?ajo?dea )+DIJEK=JA@ FH@K?JE LEIK=EA@MEJD,).,))HAFHAIAJ=JELA E?HI?FE?E=CAI B DAF-emitted fluorescence (% of basal) d control (n=4) 19 d control (n=4) 19 d + L-NAME (n=4) Time (min) IDMECJDA)+D ALA@?D=CAIB ALAIA=IKHA@>O,). 6BKHAI?A?A EJAIEJOEJDA,) FK=HOIE@AB' A>HOIEJDA=>IA?A HJDAFHAIA?AB )- JEA?KHIAIBJDA E?HA=IAE,). 6 BKHAI?A?A=BJAHJDA =@@EJEB)+DEJDA =>IA?AHEJDA FHAIA?AB)- -=?DFEJHAFHAIAJI JDAA==@5-B IK>A@JDAE=JEIIKAMAHA>IAHLA@>AJMAAA@JDAE=?AI.EC!,

12 >O)+D52);" % =@&H?/2??AJH=JEHAIFIA?KHLAIJ Relaxation (%) A % (n=12) 5% (n=19) 21% (n=6) 95% (n=6) B Relaxation (% ) % (n=7) 5% (n=14) 21% (n=6) 95% (n=6) Log [ACh] (M) Log [SNP] (M) C Relaxation (%) % (n=4) 5% (n=9) 21% (n=6) 95% (n=8) D Relaxation (% ) % (n=4) 5% (n=6) 21% (n=6) 95% (n=8) Log [BAY ] (M) Log [8Br-cGMP] (M) E Relaxation (% ) -5 5 ^ aortic (n=6) pulmonary (n=6) F Relaxation (%) -5 5 ^ Aortic (n=6) Pulmonary (n=6) Log [ACh] (M) Log [SNP] (M).EC# % + =@JDA?/2==C&H?/2,E,)HECI?AJH=F=HJIBH'@=O?DE?AA>HOI =AH=JA@MEJDABJDABMECC=IENJKHAI '# #+ 2 $!!2=# ' #+ 2 $&% 2= %" #+ F $ 2=H'# #+ 2 % %$2=2=AI-=@.??AJH=JE@AFA@AJHA=N=JABBA?JIB)+D -=@52.EJDA=HJE?=@JDAFK=HOIE@AIB,)HECIBH'@=O?DE?AA>HOI =AH=JA@MEJD# ' #+ 8AIIAIMAHA?JH=?JA@MEJD $ # 2 #BH - V 2 #BH- W 2 #BH- 2 #BH@EBBAHA?AE- =N BH=HJE?IE@A

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oxygen 21% oxygen 95% oxygen E max pd 2 n E max pd 2 n E max pd 2 n ACh (14.2) 6.44 (.36) (13.9) 6.2 (.66) 13 SNP (14.5) 5.62 (.25) (13.9) 5.27 (.29) (1.1) 5.31 (.25) 6 8-Br cgmp 131. (28.) 5.18 (.29) (19.3) 5.17 (.3) (15.5) 5.1 (.15) 8 BAY (31.) (22.4) (12.2) - 8 8=KAI=HAA=I 5, BA>HOI=HJAHEAIF, C-+ # - =N =NE=HA=N=JABBA?J B?JH=?JE 2 #BH#NOCAE6=>A! V 2 #BH NOCA W 2 # BH'#NOCA2HA?JH=?JEMEJD$ # 6DAC=IENJKHAIMAHA?FIA@B '# #+ 2 $!!2=# ' #+ 2 $&% 2= %" #+ F $ 2=H'# #+ 2 % %$2=)I=NE=ABBA?JM=I J=?DEALA@MEJD);" % F, M=IJ?=?K=JA@=@- =N HAFHAIAJIJDAHAIFIAJ

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