Estudo comparativo dos efeitos de esculetina, 4-metilesculetina, prednisolona e sulfassalazina no modelo de doença inflamatória intestinal induzida por TNBS em ratos

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Estudo comparativo dos efeitos de esculetina,

4-metilesculetina, prednisolona e

sulfassalazina no modelo de doença

inflamatória intestinal induzida por TNBS em

ratos

(2)

Estudo comparativo dos efeitos de esculetina,

4-metilesculetina, prednisolona e

sulfassalazina no modelo de doença

inflamatória intestinal induzida por TNBS em

ratos

Tese apresentada ao Instituto de

Biociências da Universidade Estadual

Paulista, como parte dos requisitos para

obtenção do título de Doutora em Ciências

Biológicas, Área de Concentração:

Farmacologia.

Orientador: Prof. Dr. Luiz Claudio Di Stasi

(3)

FICHA CATALOGRÁFICA ELABORADA PELA SEÇÃO TÉC. AQUIS. E TRAT. DA INFORMAÇÃO DIVISÃO TÉCNICA DE BIBLIOTECA E DOCUMENTAÇÃO - CAMPUS DE BOTUCATU - UNESP

BIBLIOTECÁRIA RESPONSÁVEL: ROSEMEIRE APARECIDA VICENTE

Witaicenis, Aline.

1 Estudo comparativo dos efeitos de esculetina, 4-metilesculetina,

prednisolona e sulfassalazina no modelo de doença inflamatória

intestinal induzida por TNBS em ratos / Aline Witaicenis –

Botucatu, 2010

Tese (doutorado) - Instituto de Biociências, Universidade Estadual

Paulista, 2010.

Orientador: Luiz Claudio Di Stasi

Capes: 21006008

1. Intestino – Doenças inflamatórias. 2. Produtos naturais. 3. Colite.

(4)

Estudo comparativo dos efeitos de esculetina,

4-metilesculetina, prednisolona e

sulfassalazina no modelo de doença

inflamatória intestinal induzida por TNBS em

ratos

Tese para obtenção do grau de doutora

Comissão julgadora

Presidente e orientador: Prof. Dr. Luiz Claudio Di Stasi

2º Examinador: Prof.

a

Dr

a

. Clélia Akiko Hiruma Lima

3º Examinador: Prof. Dr. Adair Roberto Soares dos Santos

(5)

Auxílio Financeiro:

(Fundação de Amparo à pesquisa do estado de São Paulo)

(Processo: 07/54516-7)

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? ! )9 *+& =

? ? !" - -

"= G )'99 N H"- 0*+

= ? - !"

53 ? ! - =D =

= 4 5 !6? ! )

*+

9 )* ; G22;?9

(33)

=- 3 -

" +

==4 3-

9 A- < 9+ &

56B( )9 I6 A6* ;k "=

= =),YFNL -M*+

. M 1 .

3 3 4 ? "

? ! )> 0*+ &5 = ;

3 4 4 ==

)(8 N H"- *+ $ ? ; ?

9! ? J"6?K == - ]A ;

" ? ;

? = <"+ G;3 I

3 ? ; 9 G22 9A

<= 3 4 )BFN,F-1*+

>;3 4=

? ! A

3 )99Y8 N B- *

+

7A " " !"

? ! -

( )

* )C8 1*+

( - 5 3 4 34?- 5-

- - -( -( -(

(34)

9

$ D D 3 4

34?3! - ;D??!"

" 3- D <= !6? !

", 4 ), *+

&-"3 4 3 A"5

- ? " 4

[-4 ? ? !)99Y8N

B-*+

> 3 3 4 34? 4 "

9 3 3! -3!

3! = =

? ! )' -*+

. M 1.& && -

> 4 3 "

3 5 +? 44?"

3?D 5- <"3 4-34=

" ;=A=

! A")9Y DNB"-*+?3 =4

? 5 ; " 4 =4

-]^ , + " 4 4

? ? " )99Y8 N B- *+

^ !" 3

, "" <")'8

M*+

U ; G22

3 ;4;k

(35)

H- *+ ? 3 ? ? ;

3 " ? ! -

5 <" ;k = )BF N

,F-1* !6? !-; =D-9

D ? 3 - "

, -4? )H-1*+&

3 4 ? !"

== -"= )'? *+

) :-: .

> 4

5 Z <" D

" " 6= =+

" " " "-

?"=?^+( -;

" 6 "- <" 3 4 6

"- Z3 ? ! ==

? ! ),Y -*+

G= 5 ; 4

=- ^ ? D- ; 4

=)(=F-*+

% ? 6- D ! 4

?3?-

;-] )= * )= *- 4

5 ; " 4 - ? ?- 3

5- <" 3 4+ > 34 5

" 3 - "A5 ?+ >

(36)

11

;3 ?< +RA?4

?-D4( 9-;" ?

")(-1*+

> 4 ?

!" < 44-]?4=

" 9-;

;)(=F-*+

> 4

?"< - !?-!?"4 8+RA

; <? ?! H- ;

5 <" <? )(- 1*+

D ?! H- ?- "D

- ; 4 " ?- 4

! = 65 " ?

),F-*+

G?^

== ? + G ? "

- 4 3= ? 4 ? !

! -?! -!?" !?+4-

^ G0m Gm !5 4 H

)%-*+

( ;

G22+ (4- ;- ?! H-

? g "),F-*+

'36- ; ==

L$ G + % G <= ? )H%/6d*-

4?)2%/6f*- )2C*2C62C6 -; L$4

(37)

;<39 G;L$)n= F o

*+

' 3 )* =4 ?

5 9- ; 9 A ?

?! H9 = H9- =

? ;+

> == "

? ! 4 = -;

= !6? !- ; < ; "

=A ? ! )(8 0*+

9 ; < ?

!-= ; ? ? " 4 ? !-

- " 5 5 ), N

L8-1*+

> 4 3=

!"" ="? g3 4 <"

= -?[ I),Y -*+'36

; I ==

3 4 " 9= ==

? ! - 4 = ;

9+

5L

&5 = ; ; ? "4 A

== ? ! -

G+G ""4

" )="9N(=-*+&

(38)

13

==-; = A

"16l) N' 3-*+

- " %>G )< "D <* 34

LGM )< M = * 4

"== G22- G)Bp

*+

&5 1=^"%>GLGM =

L-L?+?5 "4

9-4 36 ; A 9 <

)C('* 3 3 4 " = 3;k = %/6q

)>" *+ Bp )* = ; = ? 4

3= G A ? D

= %/6q C('- ; G " <+

&4+=D;

? ;%>G- J 68K)HCL*

A= 9 !"- ;k =

-D 5 =A"

= ? !)29 *+

U <= ; " %>G HCL "

9 3 4 = +

- = %/6q - = %>G

39 - 3 4 "

- A = = -

!6? !-;

== G)29 *+

4 - ? "- >H% ) A

"I* 34 9 'C0M- D 2 GM- 34

G)( 8= 0O 1O7

(39)

53! A =4 3

)( 8= 0*+

> " 34 G4%CL(?<

% )299 *- = ; 5 A

"? 3 DH%

" [- 34 3 G

)7 *+ ' ? 34

9 " "

)299 *+

> ? "4 G22 4

5 ^

<

?"- 3 4" == =

+

@ & E

> ? !"; =3=

? ! =+ > 5 = 4

? !" 3 - ? = 4

= 5" )&L>'* " )&L%* 3=

? !+

&4 = 5"- ?

3 5"- 4 =- = g

4 9-44 ?

5 + > 3 =

" 5 4 5 =)LD *+

>=3=? ! .

(40)

15

9")B>*9 )>*),TD 1*- !?

L$! G-? =)T

N T9 1*- ; ]A A + &

; =

=-5>6 ? Z !?

?-4 ==3DA;[+>

B> ; !?-

! - = ?! H- "" 3D A

;[)TN7" -*+

> ; = 3 < 3 "

5 <- " ;

? " "- ; "

3;k +H34=

" <- " ^ ?

4+/ -;A4? =-6

" < < ? ^

"4 - ;k " ==

)L *+ & Z ] ?

I6 =) 81*+

? ! D

!? !?"- ; ; = 3 =-

; 3 4! +

& ? = " 4 ?"< ;

3 4 = 5"- 4 3 5- ;

] ""A9[+"

D 5<- =-

A=== )G"

(41)

G ? -4 A=? 5 <=

=G22 ] 5 9)TN7"

-*+ & - " ; A

= ?

5 !" )T 38 0*+ )*

3= =? !

5 =- ; C96 F )1* = 5

^ <= 5 = 3 !

?+

N&

3 5 - ? ?A 3= ; 5=

"= A? !+

( - 4 ; 4 ?

<=== ? !+(

4 ^ ? 5 ; ?

DA 6DA )TN7" -*+

& 5 6DA < ;

? -=" 5

-"D-= &- ?-"D-=!+ - " 9

- D ]A 4 = 5 6DA

; " - <-AZ" <

d6 ?-333;)TN7" -*+

> ?DA A 4<?

? =+ &

D !5 )'>G*- )H*

(42)

17

&^?!" = D 5 4

3 35 <=4 =5"

"+c;3 ? 5 !"

G22- 3 "

? ! )T *+ G ? -

;; ?^ GL$

=H(> <+

?? N G )M* 34 <=

-" D 3;6 G22+

35 -3 D=-]

= 5 !"- DA

DA - 34? ? !

)%NL86,-1*- 3";

=?A+

7 .K&=

D ? !

- " Z ?A- ;

- " !- -

)i 6/9-* 3!"),+*+2?D

<= G22-;?A<=

4 ? !

-5-?53- ; D-4

5 ;9?A5 +

% - D

" !?;

(43)

> = A M6

< )M6'*- ;

?D-3D- D D+ & ?A

?AM6<" D+

?D 4 ?A 9 =

=-==),+-*

= G )%= N B- *+ > A

g = ? !- = 3 <

"-3g 5 )(-0*+

?D D

=- 4 = " Z ? ;

+ & ? 3- A 5-

; <= 4 ?- 4 - ^

4"- ?3- 9!- )L? N C"- *-

; " ? =

?D- ; 34 ?A+ &5 34 =

^93-;9 - --

?4 ; ?A

)(-0*+&-? 9 6? !5)C *

? ) *

?A+

> " ! ! ?A ? ? !

D = =

= 9 )%= N B- *-

?A D +>"

?A " 4 !

; "+ > 5 6

!=?<;"Z4"6

(44)

19

5- 6- 3 < - (6

%/q ),5- M*+ G= - " ! ? ?

=A4==? !5=A

-;- =

!5 < D<= )%>'* 5" )>E6* )L"- *+

' = ? ! D 3 =

? " ? ! =

-3 = ? ! ) N H&#-34;- 0*+ &

?A"

!?"A?-32C6! -2C6H%/6d?! -??

" )'9N(8-1*+

G= ? " !

- " - "

A G22- == "= ?

-; 5B()L"-*+

G ? 3= 3

< ? !- Z3 A63-

^ 9A = Z6;4

-9"- 3 - == 9

- ; ;k ! = ?A+

/ 34 " H2

== Z 4 - 4 ^ 3 "

!- 3 ?^ 9A )%= N B- *+

4 ? 3=-

? "Z -;?3

= <??D)'99-*+

3 " !

=G22-? -4 3 =G22-?-3

(45)

> D 16 34

D G22- ? =- 3

GL$)L"*+

-; 4 3! =- !5- A"-;

=3< G%+( -^4

?A )L" *+

? -

? 5 !- ==

3 - 4 ==

?)%=NB-*+

> 5 -

!- 34 D G22+ > 5 "

3 ? - ? - ; 4

A< G%-=?G%?

; - 4 H+ G ?

- 9 ? 5 ?3

)L"-*+

> " 6H%/- 34 3!"

?A- D G+G

D ?53-3 D3)iNB-*+&

?A ;4- 6H%/; ?A

G- 3 ?<

)%=NB-*+>

DH%/-4 = DH%/-4 H !?DH%/-4; =

)(3 *+>? ==

- ?^- 3 ^

? ; - ? - 4-

3 ?3- 4 ^ ?A )%= N B- *+ 4 ?

(46)

21

I 3!" )i N B- *-

- ; =3D

3+

= M6'- !-

3!" 3?< ? !

?A )%= N B- *-

? =

A =- D

? ! [ " < )i 6/9- *- ;

?D A 3 = = ? !

Z -="=+

1- & & :7. K :

G ?^ - = =

5 4" 3 = ?A

= G22+ G ? - Z ? 34?

? 5 5

- ?=! ; )'A9D , +- 1O

'A9D,+-*- <)A=D+-*-)D

+-*-)> +-OA=D+-*-)+-*

9 ) +- *- 4 = & )DD +- *

)DD+-*+

> ?=! " ?!

35 - 3< D ; ?

= 5 )8 N '- MO / N L"-

0*-4 3 *-4; D ] =

(47)

), N TY- *+ & ? =- ?=!

D? ? 9=? ."

=- 3! +

6 = 5 ;<

?=!- ; 3 95

4 =?!" -?

3 !

? ! +

" ?!

- ?" 3 4+

H34 3D- A 69!56

I- -63D-

+& =" =

A I 69!56I-

; = 3 D ? = )B N

(FA-1*+

?=!- 5 =A=

- ? 3 4 3A- ?

=3!"+"]A?

= ?!"- 5- ;

= - 9- 9 5 = )B N

(FA-1*+4- 4 ; " 695

?5 )-669!5616 56*- )1-66

9!56* 06 )1-669!5606 *

3 5 - 4 I !5

;<?)T NL9-*+7A=<

= " = ? ! 5

-3 = -3 -3 DA )L=

(48)

23

$ D G ' +- )- 0* D =

" ? ! -4

)/< &*+ % - 3=6

= "- Z D

6A < )G ' +- *- ?

= = 5 " [ D A

3D?[)H% '* )G' +-0*+

3 5 - 6 3 ! =

= 5 0 5 < D

?6A !3 3 43 ) 3 5*+ &

; 06 =

3 !Ml)2M*5<-=1-M

1-µ

,

= ) +-*+

G ? - 06 = <

9 = 5 )(FA+--'9

M*- 4 < = ? ! 3 =

5"5"6 =)'8= *+

& 3 I- - =A

?!" - = 5 ?

===A=-? = +

& ?,G)<*

D63 69!5)6 B(*-=G((B)-69F66

9D* )C *- " G% 5 = )T8

*- 3 95 = =

G% B>)T *-35 5

=)C *+4 =- "

9 56 B(-? !?

<= " )C *O 3 < 35 - I

(49)

)>8 M*- = ) 3 *+(

? . !?" ?!

)C" M*- 3 )" +- *

)(8 *494)i"

+-1*-3 [ ) 3 *

4 '' I )T8 *- ?3

3 <= H/6e ),3] * 3

5%/6q = <6)(6*)( *+

06 = 3 ?

3 I < A 9[ " I A

)B] *- = 5 Z - ?

) M* ")

*+

3 = 5 - 06

? = = ? !

5 + % = =

= < ? 5

? ! D A 3D?[ )H% '* -

?"[+& ;3

I-M"T"- = ? ! 5

= A 5 ? !

3 I) +-5*+

&

= = ? ! -

; "-4?A

= <- ?A ? D G22

9- ?D - ? ?

?!" ?A? ! D

(50)

25

<&.N .&& 7. K .&-O& .%& 1O-.7PQ0#)

& 5 ?

;<+ 4 36 9! ; G22

3-;? <"

I !- ; 4 ;

!"+

>4 3H% ' Ml-;

"J3K-; ;AH% '-

9 - ; 6 3 I -

< -D !")=

*+>H% ' === ?

=- " ! -Z

"? - ="

+

&5 4; =

=? ! - 4;

? ; G22 9 )R]

0*+

B " - ? D H% ' 4 D

Z - 4

" )/"4 1*

--? 4? !+ ; -43=

= D 5 )'A96B"

*?? )'A9D,-0*-4 5

!6? ! H%/6d )'A96B" *

(51)

'"' 3 -)*-! -H% '

4 4 H 4 <" I

!+9 H% '=2C6

4H9-;D 2%/6f- =D"

!?"D !6? !-H%/6d-2C6e2C6

1 ; ? ! )/" *+ 4

-=A ? ! == "

&- 35 - 00 )V9" *+

DH% '4 5 ?

- ; = ?

A <"<?)' 3

*- 3 = = - Z

G =- ==

== ? !+

Į

ȕ

Ȗ

!"

# $

!"

%

ȕ&'

Į

ȕ

Ȗ

!"

# $

!"

%

ȕ&'

- / &&-& & 7. K ..0#)

(r4 <"O((r((FOHr4HO,r!?"+

(52)

,*7

(53)

+ = ? ? ! =

< ) 06 *- =

=

? ! O

+ = < )?A

*? ?A? DG22

9- ?D - ? ?

?!" ? "- [ =

DH% '-= ;"?A ?

DO

+ = = ? 0 3

I <- !- ! 3;< ==

? ![-??

O

0+=?? 3

-? !-

5 +

(54)

%%<

(55)

L 93 "- 4

$- - ' ( ? D

= ? ! + > ? 3 4

G /"- 2 - $- M

< 5 + & 3 4 3

±@69+> ?]

5- "- A5 1 5 ?

(C3'+2+A" +>

? D 5 = U

&5 2 )( +00#&&*+

)-1R &2N .

& )1-6956*- 06 )1-695606 6

*-?D- /"-?3

'"-3D l-

? 5 35+

(56)

29

- $-- ?-= & -1R

. & & & & -. 4 9.-. 4 &. -.7 . O 7 -& &7. K .

% ? " ? D " "

5 .

A. 3.

B. .

C. 6 .

+ M"T"

+ 06 M"T"

D. ?A.

G+ "T"

G+ ?DM"T"

S NH N N O O N O OH OH '?D O O OH CH₃ O H CH₃ OH ( O O O H O H O O O H O H CH₃

(57)

? !?D4 ,

)*-;?^+>?3 <

]] 0 9 I "A >

?<+

&"-?D ) [*-M

"A-0-16 3D?[)H% '* Ml==+

? D ] 4 )I *- ; ?

DI 4 I+>?

335 9 4

+>"3?3

3 H% '+

> ? = D6

?A"-?Dg1--0-09 +>

" 33 l=+H

? 09!? !+

(58)

31

. & & & 7. K & -. 4 9.-. 4 &. -.7 . O 7 P & & 7. K .

> ? 5 [

=?;35)/"0*+

%?[=- ) =*?

9 ! H% ' 4

+ / = 5 [- " 3 )6< *-

" )< *- 0 " < )M"8"*- 06

)M"8"*- )"T"* ?D )M"T"*+ '

"< "3? !-0

-; "<

=? !+" H% ')=*

? D 0@ ! - ; ?

?@-5 6"3" =+

- G9 5 +# )" 3O < *O,)" O < *O,@ )" =O < ; 3 " H% '*O0 )" O <

- 06 - ?D*O 0@

(59)

. &7. K .

? < - ? - !

5 < ; ]<D 6 I

!-!3;<+

. K

G == 5 - ? =

? I A " .

?D 4+ % A ! ? =

!-5 !"]

A = 5 ]<D -

= M+

0 1. + 4==5 [+

$ ,L

0

M

16

']<D

B-Z

s?"?

s?<

G< ?

G < ? < ?

;"5 !

' ]<D 3 ; " 5 ! )

4 < *

!A!!?=?" "

(60)

33

?? - - 5- H%/6d- 2C6e Z

<[-; ?" ?

<=" +

. K & &K.

2 !=!?

)-M* ] g A ?

9 !"+&

??5?0l09-= A" " <- 4 ! <

- ?D 5- < ? B 6

( 9),861* +> -1[

- ? ?D D 4 ! <

+

"- ?3 g69 5

A ?!" ^ < !

4 ' 9 )*+ I 9 !" ?

3=!! ? "?!! CcY

(=++0- E+

(61)

0 1. + 4=![+

E .& ,L .

/- +$.)*%9O'=)*O,)*O'=)*

, + =, ! .)*%9O'=)*O,)*O'=)*

+G,.)*%9O'=)*O,)*O'=)*

ER K

0+2? ,.)*%9O'=)*O,)*O'=)*

M+2? < .)*%9O'=)*O,)*O'=)*

1+7.)*%9O'=)*O,)*O'=)*

)-1-

+2? ,.)*%9O'=)*O,)*O'=)*

+2? < .)*%9O'=)*O,)*O'=)*

+&.)*%9O'=)*O,)*O'=)*

&,A5.-?' 9 +

&S&L.-. K &

> Z 4 ?

4 /")/"NL38-0*+

9 !"- ? - ? 1

[ -?D D4! <

+ "- ? = A"

? g1@ B% 34g1@-M +!

?= =5A" +

! ? 3 = '9? - ?

" 0M + G <- ?

= A" + ( 5 A-

? = = D l- M + >

9 !"?= = D4! < +

A Z 4 ? D !

(62)

35

5 -5<3] ="<+/

Z Z4 +

/ 4 ! ; ; =

]Z! D=6!53

-? -?"M+> -?54"

Z 4 ! =Z +

- ./ "?! " 5

(63)

M1?-=

H ^ 3;< ? D 9"D

[- D ? 5 0M

"-]9"D A -= ?!

=+

& & && .N& Q/<

=5?" !

? D 4 TYD )0* 5

5" - =,(>4

?;;"t!59" M₊

= 5 D ?

!?-; D]<!?-;? +

# &S&&.- .

Z " ? D

4 )M*-; A35 "

D )'B* g ? 5 )''*-

3 A 3 3D! )GH% *+ > GH% D

;-; ? +

> '' " 4 D D "

%G(B+ > 'B ? 5 = - " <- ;

= GH% ) ;k 33I

0* 4 g ; " )'Bm''*+ >

?5" +

(64)

37

, &= & & && 77 . .

Z < = ??

?D4 A ' 9 )M* F

)01*- = +

&S&& ..& .&=&Q/.P

Z<[?D

4 V" )*-""?^+

? 9"D T +Ml

.M+>9" ? ?" 0@+

/ D0t3 -;? 3=

- 34?t? !+l-Mt

A 4 l- B +M- M t A 33 Z +l < A

4 0 t A" + ( = - ? D t

? ^ <- t A"

" +

> ? ? 396 M@

9+ ! ? ? ?

?"0 +t3 ?

1 ?D ? [ M+>=

,G ? M ? 5

" +

(65)

$ & .

= " < ),,(* # 6

! ? "5 ?D"?"

" +(5 D-"

- H6BM,-B+M-

-Ml H E6 , ? 9"D

( M " + > 9"D ? 3

" 90_{?5 +!}

<-9"D? ?"+L(,M 0_{+>3 ?}

= Z < ? ; ? 4 ?

)1*+ 5 )u" <*?3 g ?-3

^ 6 - " l D +l

" + G ?- "4 ? = =D M

+Ml H E6 = ? !

)'G'*-" =)'G'*-" M 6B M ,- B

M,,V+( -"4?360

9 _{- 3 " + ?- "4 ?}

L6M+> =3?

D D

?+ A ; = ) ! " #

2>G*33 D"??D D62", 7G'

=+92", 7G')(9 9*+>

?52>G+

&=&09T:E9U

> <= H%/6d 2C6e ? !

(66)

39

+-).M*=0@+!9"D ?

" 396g@ !? ?"

+M 0@+>3 ?9D?D

6@ 4D"+><=H%/6d2C6e?

8 )&C2'*-=3 ?5"" <

)L"D63D *+

. & & &&$-. 9.-. 1 &

K97. K

(=? 06 -3

!6? ! ? D 9" .

4 9)6*-!?"")L10+*

4 =A-3 ?! 3" 3v+

4 )6*- ; 4 9" 4

9-9"L10+?3 '= =

$=6&9+& 4? @-Ml

> ?4 = G,&, )G3v ? &"v

* -C6" ),*-

)M$C*-? )-M"C*l)M$C*-? 3= =)0 -M1

*-?G,&% +

4 =A?3 ?! " 3v

" "+ ! - 3 ? =

= 3 5 + & ^ 4-

!" ? = M G,&,

l 3! ) *+]

- 3 ? 4 ? 4

= + & "- )m4* ?

(67)

- ; ? ?" M M + > ?

M )%B0-C-TB>-C- 6

4 C* g0@-3] = ! +!

--; 4 ?! !?")! *-?

?"MM "? M

G,&, +

& )&, K9:7. K

? 2C6 2C6e 2/%6f 2C6

06 ? ^u,-u,-M>u,

u,+

( 2C6? D4 6+

& 4 ? 3- < 0 9- 06

^ + ! < 4 ?

2C6e ) "* < 9+ G -

3 ? ?" g + & "-

??D6@ <=2C6

&) 2 -%=- "*+> ?5

2C6)"*+

( 2C6e? D4L)5M

4*+& 4? =0

? ^ = <-

C(')M"*+!19-3 ?

6@ 4 D +( <=

2C6e ? D T & ) *+ > ? 5

2C6e)"*+

( 2/%6f2C6? D! )-M

(68)

41

= )Mu"*- 4 ?

? ^ 3 I + ! 0 9-

3 ? 6@ 4 <=

2/%6f2C6T &) *+> ?5

2/%6f2C6)"*+

#D 1.& &,.-.

(= !53 I -?D

=33? 4 D +

=346L10+? 4

= -" " E )*-"

?^+ 4 ? ?5 = )+l*

)l* g 3 +!0="( '-4

? ? ! )'G' l* +

( - 4 ? ?" M

+ & "- ? ; ? &C2' M0 +

4 =A= D 3

)29F 1*+> ?5l=3+

=3 ! ? D L" 'H6

)L9 G" *+ & 4 4

= 3!?+>Z=)'H6*

4- ? " =A=- + c

?- = - ;

9! = 'H6- - ; 4

0M+

4 = 3 )! * ? = 0

)-M 5 1_{4* -M 4- ?}

(69)

Mu = 'H6 )L9* 9 @ + & "

? " ? ; ? &C2' M0

+> ?5l=3! +

%. =

H ? 5 4 w 4

)&+(+,+*+ G? 4 ? A 7I )%>7*

" "?I+ G 4 )* ? 5

? T86" G+

G < ? E_{+ "?I < ?}

=x-M+

(70)

I

(71)

@-. &&.N .&7 -&

. / K

[ H% ' ? 4

[- ; = ! 0 9

- 6+#M+ "!-=

9 + & I

? 5 ? -

3= 3+

>

! ; " )1 =+

= - x-M*+ > ?D- 06

34 ? "

) 3*-49? < ?3=+

5 -" 5

4 0-1w-01- 3 I

5 ?- 9 ? < ?

3=+

=6

[)"*-" 00-1Mw0-0 [)"*-" =+

-w- " 3 ) < *- ? <

x+-49 = I +

>" l4-; ;

4-l+c g- ?3=-l

" - -l

- 06 ?D- 9

(72)

0 1. + = I ! ! ? )M"T"*- 06 )M"T"*-)"T"*?D)M"T"*5 "? ! DH% ' +

y>= 5) =*-3_{5 -6 [}

54w&+(+,+-_{4? E}_{-zx-MOzzx-Ozzzx}

" + H " ? ? "? = " 3 x -)"?I 3*+

5-

K

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(73)

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- . = "- 3

5 " D H% '+ G 5 Z +

? !-

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06 -?D 3"9

!! H% ')/"*+

-8 -6 -4 -2 0 2 4 6 8 10 12 14

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(74)

. K

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= -

- )= *

? ? !3 I!)=*

3 )*- ?

"< - ; <

? !+

>

=D I !- ; " + >

06 -?D

? "< 3- ; 06

34=? I!)?"

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)* 5 ? !- ; 06

< "? = ! "

9 g?D) 3*+

(75)

&

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! H% '+ 9 5 )B&*+ ,? "? ! ! C cY ( = +0- ME+ =- =O - 06 -?D-?

3+ .Q =-Q1 ? 4

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& L.-. K

Z 4

Z ;"

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= Z 4

" )/"*+ 0 0,02 0,04 0,06 0,08 0,1 0,12 0,14 0,16 Cé lu las apoptóti

cas por cri

p ta # ,. $-. Q*I 9.-. Q*I &. Q*I )-.7 . O Q+*I

- ".=Z4 3

5 "DH% '+G54w&+(+,+

(77)

. M#?-=

; -? ! =!

H% '-;3= =,(>)/"

*- ? )/" * )/" *- 4

<= H%/6d )/" 0* 2C6e )/" M*- ;

" < )3*+ &

9 "? Z "

[)/"*

Z<),G*)/"*-;k 5 =;! A3 +

> 46 - 06 -

?D= <=" [-;

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Z'B-9? < ?3=

3 I ?A?- Z,G

)/"*+

> 46 06 - ?D

= = ?? ;

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= D 5 )/" *+ c <= H%/6d-

D <= 0Ml

)/" 0*- - 9 = "? =

<=2C6e)/"M*+

=D ),,(* ;

= ,,(6 ? ? ! [ ?

" =,,(6 -;

" 3 + > ? D 3 =

,,(6 34 = ,,(6 5

(78)

0 500 1000 1500 2000 2500 G S H ( n mol/g t e c ido) # ,. $-. Q*I 9/.-. Q*I &. Q*I )-.7 . O Q+*I

**

** ** **

**

- +.=<=" [)'B*3

5 " D H% '+ G 5 4 w &+(+,+ zz

x+" + 0 50 100 150 200 250 300 350 MPO ( U /g t e c ido)

- . = = D 5 ),(>* 3

5 "DH% '+G54w&+(+, zx+M" + 0 2 4 6 8 10 12 14 16 18 20

FA (mU/mg pr

oteína)

**

*

_*

- = = ?? )/* 3

5 " D H% '+ G 5 4 w &+(+,+ z x+Mzzx+-" +

(79)

0 5 10 15 20 25 MD A ( n mo l/g t e c ido) # ,.0#) $-. Q*I 9.9-. Q*I &. Q*I )-.7 . O Q+*I

*

- = <= < ),G* 3

5 " D H% '+ G 5 4 w &+(+,+ z x+M-" + 0 50 100 150 200 250 300 350 TN F-alf a ( p g/m g prot eína)

*

- = <= H%/6d 3 5

"DH% '+G54w&+(+,+zx+Mzzx+-" +

0 1000 2000 3000 4000 5000 6000

IL-1 beta (pg/mg proteína)

- =<=2C6e3 5

" D H% '+ G 5 4 w &+(+,+ zx+M zzx+-" +

(80)

0 50 100 150 200 250 300

Branco Controle Esculetina 4-metilesculetina Prednisolona Sulfasalazina

IO

D

MMP-9

MMP-2

- . A 4 =" < ),,(* 6 ,,(6 D"?- 3 5 " D H% '+G54w&+(+,+ ! )2>G*-zz x+" + //9"

- . =" < ,,(6,,(6 D"?+ ? "? 2" , 7G' )(9 9*+ r O r O r& OGr06 -&r(/r'?D+

(81)

@-. & & &. N . & & 7. K . & 7 P &

. / K

5 ? "- [

D ? 4 [

+?! D

[- ? ! ?- )3=

9 " = 5* ? "+ &

4 A5 0 9 ! H% ' 3=

"-D - 4 +!

- ;3=5 ?

"-""! H% '

)H30*+

%5 ? ! ?[-;

I !- ? <= 3= ; 06

!-- ! + >

9 -4"? =

5 4)H30*+%"!

H% '- ? <= 3=

5 06 =

[;

" + % ! =-

- 06 - =

5 ; "

-? -?A -? ; = 9 9

"? = I ! =- ! =

(82)

5-

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-)"T"*?D)M"T"*5 [=? DH% ' +

y 5 ) =*-3_{5 - 6 [ 5 4 w &+(+,+}

4? E_{-zx-MOzzx-Ozzzx-" +H"}

(83)

%=-?3=

" ; " 3

)6< *- ! H%

'-;?"" )/"

*+ H = "

- ! ? !+ >

= < ?D = "

- ? 3-9=?

< " +

>" ="9;

"3)6< *-"

+ > 06 =

"9 ?A?-;

3= ?" + >

?D- " 5 - !

"9 ?)x-M*

" + > " ? "

M----0+&;3?D?

" - 0 1 )? <

?"*+

(84)

20 25 30 35

1 3 4 5 6 7

M

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di

a de consum

o de ração por ani

m al (g) # ,. $-. Q*I 9/.9-. Q*I &. Q*I )-.7 . O Q+*I

19 21 23 25 27 29 31 33

2 3 4 5 6 7 8 9 10 11 12 13 14

M

é

di

a consum

o de ração por ani

m al (g) X) + + + +

! " + ! " +

M

é

di

a consum

o ração por ani

m

al

(g)

X)

- !.=43 5 ?

[- - " ! H% '+ G 5 4 - "+ G H% ' 0P = )]]*- +

X)

(85)

+ ! + ! + . &

& Q

V # ,. $-. Q*I 9/.-. Q*I &. Q*I )-.7 . O Q+*I

**

X) -5 0 5 10 15 20 25 30 35 40 " .

& Q

V X)

**

9+ + + + + + +

" "

. & QV X)

**

2.

. K

,.&

- ". = = 3 5

? [=DH% '+G H% '0P =- + G 5 4 )( ? 6( *E(*w&+(+,+zzx+" H% '+