. . . , .

...

. . . , .

..., .

. . . , .

-

 

 

 

DCV -

CGRP - -  

 

(CGRP) , , 80- . , , 쐍 , , , , - , - , CGRP.

CGRP 37 , , , , , , [8, .265-271]. CGRP - (.. dense cre vesicles, DCV), [14, .14294-14298]. DCV . , - , , , , .

, CGRP ( ) . , DCV , , . CGRP [12, .432-438; 10, .10].

CGRP - . : 1) 7- , G-, 2) - CLR ( . calcitonin receptor-like receptor), 7- , 3) , G-- . - RAMP ( . receptor activity-modifying protein) [12, .432-438; 20, .1881-1886].

-   CGRP , , 핐, [19, .24994-25000; 27, .394-403]. CGRP . [17, .934-939] , , , [24, c.345-350; 25, c.689-695]. , .

, :

1.        GRP , rCGRP   . ) rCGRP 1 - 1; ) rCGRP rCGRP CGRP- - rCGRP8-37.

2.        , CGRP CGRP . DCV ( CGRP) .

CGRP

CGRP (calcitonin gene-related peptide), 37 , . 쐍 /CGRP : CGRP, CGRP, α β; α-CGRP CT/α-CGRP . CGRP, β-CGRP, , [13, c.43-47]. CGRP, α β I II , 1 3 , , . N- 2 7, CGRP, CGRP8-37 (. 1). α- (8-18 ), β-, 10 . β ( Pro28) γ- ( Pro34) CGRP. , , , . , CGRP CLR (calcitonin receptor-like receptor) 2-37 ECLs (extracellular loops ), 8-27 N- CLR RAMP1.

 

: a-CGRP (human) Ala-Cys-Asp-Thr-Ala-Thr-Cys-Val-Thr-His-Arg-Leu-Ala-Gly-Leu-Leu-Ser-Arg-Ser-Gly-Gly-Val-Val-Lys-Asn-Asn-Phe-Val-Pro-Thr-Asn-Val-Gly-Ser-Lys-Ala-Phe-NH2

CGRP

: CGRP (rat) Ser-Cys-Asn-Thr-Ala-Thr-Cys-Val-Thr-His-Arg-Leu-Ala-Gly-Leu-Leu-Ser-Arg-Ser-Gly-Gly-Val-Val-Lys-Asp-Asn-Phe-Val-Pro-Thr-Asn-Val-Gly-Ser-Glu-Ala-Phe-NH2 (Disulfide bridge: 2-7)

CGRP

 

: CGRP 8-37 (rat) Val-Thr-His-Arg-Leu-Ala-Gly-Leu-Leu-Ser-Arg-Ser-Gly-Gly-Val-Val-Lys-Asp-Asn-Phe-Val-Pro-Thr-Asn-Val-Gly-Ser-Glu-Ala-Phe-NH2

CGRP8-37 ( CGRP-).

 

. 1. CGRP, CGRP8-37 , CGRP-.

 

 

CGRP : ; - . CGRP , CGRP : , CGRP , , [26, c.163-167; 7, c.967-969; 19, c.24994-25000]. , CGRP 쐍 - [19, c.24994-25000]. , CGRP , - [19, c.24994-25000; 6, c.1061-1073; 5, c.20-31]. 30- CGRP, - - (CGRP) .

 

CGRP

CGRP (CGRP1 CGRP2) [26, c.163-167; 4, c.729-732]. CGRP1 CGRP8-37, CGRP2 . CGRP, , CGRP, , CGRP , , CLR/RAMP1 CGRP1 CGRP, CGRP2, , [11, c.143-145; 4, c.729-732; 9, c.1885-1898].

CGRP CLR (calcitonin receptor-like receptor, GPCR), , , RAMP1 (receptor activity-modifying protein) RCP (receptor component protein) [16, c.333-339; 29, c.476-483; 5, c.20-31]. - CLR RAMP2 RAMP3 2 3 , , CGRP. 7- CGRP CGRP8-37, CGRP. C CGRP, CLR, (BIBN4096BS) (0974) [26, c.163-167; 29, c.476-483].

CLR GPCR -, 6 ( 6), GPCR - ( ) DRY , GPCR, Arg151 TM2, His155 TM2 and Glu211 TM3 ( A GPCR Asp-Arg-Tyr). CLR Gs RCP. CLR Gs: (R173 ICL1 (ICL ) 249 ICL2, , , G-, L169, V245, W246 W254; , / R336 ICL3 . - CLR 8- (eighth helix, G388-W399), ; Gαs. RAMP CLR ( RAMP1 CGRP, CGRP), , N- RAMP, - (10 /) RAMP1, CGRP. RCP , CGRP- , β- [28, c.20120846; 29, c.476-483].

, CGRP [7, c.976-969]. , CGRP ( ) , . , , , (, ) [7, c.976-969]. , , , 핐, [27, c.394-403].

CGRP . [4, c.729-732]. , , , PKA [3, c.361-378; 29, c.476-483]. CGRP 2 [18, c.582-588]. , 핐 (3-4 ) - CGRP, . CGRP , .

 

. 2. CGRP CLR, RAMP1 RCP ( , CGRP ) [29, c.476-483].

 

CGRP - CGRP [25, c.689-695]. , - CGRP (1 1 )   , . , .

, CGRP . , DCV - , , . - , CGRP . DCV [21, c.7799-7806; 22, c.4477-4481]. CGRP .

 

,

- (m. diaphragma n. phrenicus). 129/Sv 20-25. , . . , ( ). .

- ( 5 ), (95% 2 5%O2) ( : NaCl 135.0, KCl 4.0, CaCl2 2.0, MgCl2 1.0, NaH2PO4 1.0, NaHCO3 16.0, 11.0; pH 7.2-7.4), Sylgard . (20-22º). ( ) 5-10 .

(Pyrex) , 2,5 Cl ( 10-15 ). . ( -50 ) (). - L-Card -154 PowerGraph, MiniAnalysis (Synaptosoft). -70 . 5 , 20-60 . - .

:

rCGRP (American Peptides) 1, 10 , 100, 1

rCGRP8-37 (American Peptides) 0,5

(Enzo Life Sciences) 0,1

-20º , . . ( ) 0,01 % v/v.

C

, , ( ), . t- . 0.05 (n - ).

rCGRP -

rCGRP - - - 1 , 10 , 100 1 . , .

1 rCGRP 1 -74,010,27 (n=10). , - -70,884,89 (n=17, p>0,05). 1 rCGRP 1 1,370,08 1,760,16 (p<0,05) , 26%. 1,150,06 2,030,11 , . 1 ( 1,330,08 2,460,17 , , (p>0,05)). 0,620,07 , 1 rCGRP 0,440,07 (<0,05), , 10, . , CGRP [29, c.476-483; 1, c.335-339], CGRP ( ).

10 ( -69,693,91 (n=16), - -66,672,21 (n=18),  p>0,05). 10 rCGRP (0,630,13 , 0,690,28 ). , ( 1,290,06 , 2,10,1 , - 1,260,08 , 2,10,07 ). rCGRP 10 - 34% ( 1,10,08 1,480,16 (n=11, <0,05)). , 31% (. 3).

100 (-70,884,89 (n=17), -66,672,99 (n=15), p>0,05). (0,620,07 , 0,550,04 ). 1,150,06 , 2,030,11 . 100 1,170,08 , 2,890,65 . . 100 rCGRP 1 51% ( 1,380,06 2,090,14 100 , p<0,05) (. 3).

rCGRP - 1 - -68,132,92 (n=16). , , -72,863,91 (n=14). , (0,630,15 , 1 0,460,03 ). 1,260,11 , 2,10,26 . 1 rCGRP ( 1,260,05 , 2,010,08 ).

. 3. ( %) rCGRP ( )

, 1 rCGRP : 1,100,08 1,740,13 (n=8, p<0,05), 1,790,19 (n=8, p<0,05). 1 60% (. 3), (1 ).

, rCGRP 1 -10 30%, 100 -1 50-60 % (. 3).

rCGRP () , (. 4). ( ), , . ( ) 1,5 hCGRP  [1; 335-339]. hCGRP, , CGRP CGRP, .

 

 

. 4. (n=17) rCGRP 10 (n=18) 1 (n=16)

rCGRP

. rCGRP ( 0,1 ) CGRP. 29 , 8- 37- , .. rCGRP8-37, N-, , .

rCGRP8-37 0,5 - 60 . 2,150,10 (n=16), rCGRP8-37 (0,5 ) - 2,360,14 (p>0,05; n=13). rCGRP 100 ( rCGRP8-37) - 60 , 2,190,13 (p>0,05; n=12) (. 5). rCGRP8-37 rCGRP rCGRP8-37. , rCGRP .

, rCGRP , hCGRP [1, c.335-339] , CGRP8-37. hCGRP , rCGRP (1 1 ) , . hCGRP rCGRP , , CGRP , CGRP , CGRP .


. 5. (n=16), CGRP8-37 0,5 (n=13) CGRP 0,1 CGRP8-37 0,5 (n=12)

(0,1 ).

( ) CGRP   . , CGRP . , - , , - , , , . , . CGRP [25, c.689-695]. , , . - . , , , CGRP . - , . , , , , [2, c.90-97; 23, c.368-372].

- 0,1 ( ). , . , (0,1 - 1 ) rCGRP, 1,350,11 (n=18) 2,070,18 (n=15, p<0,05) ( 54%) 30-40 . rCGRP (. 6).

CGRP, , , , CGRP.

 

(0,1 ) CGRP- rCGRP8-37 (0,5 ).

, , CGRP- . 1,720,12 (n=15). 20-30 CGRP- rCGRP8-37 (0,5 ), , , , 0,1 ( rCGRP8-37). CGRP- ( 1,970,12 (n=21, p>0,05)) (. 7).

, , CGRP, , .

. 6. (n=18) 0,1 (n=15)

 

. 7. (n=15) 0,1 CGRP CGRP8-37 0,5 (n=21)

 

1.                 CGRP ,   ;

2.                 rCGRP8-37 0,5 rCGRP , ;

3.                 54%;

4.                 CGRP   CGRP8-37   .

 

 

 

 

 

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