Kowsar Medical Journal Vol. 15, No. 4, Winter 2011 Pages: 189-195 % :!"#$
! " #$% & ) * * + ,'" &-. (Crocus sativus) ' (
* PhD PhD MSc PhD * #$ #% ()... !" #$ #% #% +,) $ +- /0&'$( )* #$ #% 1% +% - - +- 2 3 $ 4 )*
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
Inhibition of pain and inflamation induced by formalin in male mice by ethanolic extract of saffron (Crocus sativus) and its constituents; crocin and safranal
Nasri S.1 PhD, Hosseini S. Y.1 MSc, Sahraei H.* PhD, Zardooz H.2 PhD
Applied Neuroscience Research Center, Baqiyatallah University of Medical Sciences, Tehran, Iran٭ 1Department of Biology, Faculty of Basic Sciences, Tehran Branch, Payam-e-Noor University, Tehran, Iran 2Department of Physiology, Faculty of Medicine, Shahid Beheshti University of Medical Sciences, Tehran, Iran
Abstract Aims: Previous investigations have confirmed the anti-inflammatory and analgesic effects of aqueous-ethanol extract of saffron on formalin induced pain in small male mice. Since there is difference between ethanol and aqueous-ethanol extract of saffron, this study was performed in order to evaluate the effect of ethanol saffron extract on acute and chronic pain in small male mice. Materials & Methods: This experimental study was performed on male NMRI mice which weighed 20-25 grams (6 in each test). Drugs were administered 30 minutes before 2% formalin solution injection. Then the animals were evaluated for 60 minutes in 5 minutes intervals. Inflammation was investigated in minute 25 by Mercury immersion method. Data were analyzed with one way variance analysis and Tukey's test by SPSS 9.01 software. Results: Ethanol saffron extract showed anti-nociceptive effect in 5 mg/kg dose for acute phase and in 10 mg/kg dose for chronic phase. Naloxane, Dextromethorphan and L-NAME inhibited the extract action in acute phase as well as chronic phase. Safranal showed anti-nociceptive effect in acute phase and Crocin in chronic phase. Saffron extract with dose of 2.5, 5 and 10 mg/kg controlled inflammation 30%, 66% and 80% respectively, but Safranal and Crocin didn’t show a similar effect. Conclusion: Ethanol extract of C sativus exerts its effect on the acute phase of formalin test, which is possibly caused by Safranal. Probable stimulation of opioid, NMDA glutamatergic and nitric oxide dependent pathways leads to pain inhibition in acute and chronic phases. Keywords: Ethanol Extract of Crocus sativus, Analgesia, Safranal, Crocin
ﻧﻮﻳﺴﻨﺪه ﻣﺴﺌﻮل: ﻫﺪاﻳﺖ ﺻﺤﺮاﻳﻲ. ﺗﻤﺎم درﺧﻮاﺳﺖﻫﺎ ﺑﺎﻳﺪ ﺑﻪ ﻧﺸﺎﻧﻲ [email protected] ﻓﺮﺳﺘﺎده ﺷﻮﻧﺪ. درﻳﺎﻓﺖ ﻣﻘﺎﻟﻪ: 6/2/89 ﭘﺬﻳﺮش ﻣﻘﺎﻟﻪ: 89/6/15 &' !M V&$*@ &C % / (G8 ,- -./ & &' & C/ m 4z %& . !* - V*8 5 : 1*7 R#G % Q6 V&$ (- &# % ! "#$ %& '(#) *+# ,- -./ 0 !M ^ A ` -./ " .* &8 +0Y !&V * 5- 4-# & . 12 .[{] V&$ Q R 5 ^ 8 &' 4 %& % % !& *!* #67 % * / " &: &' ! V !M &C %&I ` -./ 9 ( [= <] &8 9 : ;&# & " / e '6 % V&$ | V&$*@ % 7 " .& ! %* ? : *@ % /> !M ^ A ` -./ " .[ Vol. 15, No. 4, Winter 2011 ,-, (# ')* ( + !" # $%& ' . 4$ " A & 4; . @; K#>o 4W . 4$ % " (L. W L. ) A " J H . #>o 4W 01& . !* +, "- "! !. & )"! '( ! "! #$& .!W # ;"! #>o 4W . 4$ ">$ ? = 234 56 7 289 : * ;" < "D$ R p :! # (' * + ,-$ ! " A B&! "! 3, @; < A; . !! I;"J * qR * * !3& A &W & K3$* E> " "D$ C- # . "6 !. & )"! '( ^F gW A .[ Kr] &" F gW A ": CG A & "! "D$ A; . ) "D$ "!F KM K Kt I;6! "! G F 01& . I;"J (n=s) G M K . "!F L. , * . I;"J G 63:CH! A \ +dG # . "6 ) CH! "! KI;"J * 0F u Mt *&"! -O! "D$ A; . #&! NOH "D$ L. F !!"6 * 0F ) * K G F +dG A \ .! S" & J P * CQ4 OR R BO! (v\- & L. u/s) ) _dG * "! G .# . "6 &" !" (U V;"W X; V . W) HPLC 0F Q 6! u A I;"J J #_a6 A; "! .[ Kr] + aQ K"D$ ,*! .! "D$ A; "! & %[ !) * WG Z; .# . Ld "! ; A & I;"J * .4 . "6 !3& !" A \ & J ]& # 2) % 3 4 % ) ./ )$ 0 1 K(; `!_) 3&A " K!3& !" ^,"! : + $&%'/"5'$ 6'( 0 )-# L-NAME K; 9" ,A & 4; K; " , " a9! , . mQ &" "P4 : U; `_B &) ; 9" ,a9 (&C A 4;c"b bL) G . M * K3$* "D$ * !"! w F +dG CG A & "! ,"! _, .! (< "D$ # ;"! * C>6 Q 6! u G * . @; . !3& (,"! Q) 63:CH! ; (A " ) );* ": L. W (L. W L. x) a9 K(L. W L. t) 3$* A & "! A;* * 0F J 3$* "D$ .4 . "6 !3& !" # ;"! (L. W ) A & KL! . .!9 # ;"! ,. .# . "6 !3& !" 63:CH! ": CG p &" "P4 .4 . "6 * !" \ Q 6! ' !W 63:CH! ; );* ": " A & #_a6 , "! 49 * C>6 Q 6! u G * . @; K . , . "R .!9 # ;"! M) " a9! K(L. W L. t) 3$* "D$ # ;"! * G "! !"! &" : ) A & K 4W . .!9 # ;"! (L. W L. #$& N; G KS" A; "! . !3& A * S" "6 * !" \ Q 6! ' !_ # ;"! (L. W * 0F V CQ4 BO; * e < I Q< f * C>6 "D$ !- _$ *4&;aW@; +;J "R mQ .4 . d4F g9 %[ A < ? [ Ke < !9!$ +;J ."R W L-NAME "! * !3& 3$* CH! "! A I;"J * 0F : h G . I;"J S * . @; "P4 A; . &" K#& *4&;aW@; "6 & '*'*' !\ ]h. a9hF 04) * ^P3< L. t) 3$* "D$ # ;"! * C>6 Q 6! u G J P * Q 6! j "N; Q 6! ( , 4 . .!9 # ;"! (L. W L. ) L-NAME K(L. W ^4; . V &" [j] S" !"! ' !W # ;"! (L. W ) A & J 4W . G F d4F ! >\ P3< A; ;* "! )"! =( ;* .4 . "6 * !" \ Q 6! I;"J * C>6 Q 6! ' K!3& !" ,"! .! ;! G" ": #&! $hy : 7 28 9'(* . I;"J G 63:CH! ; );* ": A C < ;Jd "P4 . !"& " \ z<±A B : K# . "6 !3& !" "! N; * m 9 !" "! 9 * K >! yN; 0;" J * * $hy . V #;$" ,I;"J A Q 6! ' * . . P "! $hy !"! 4\ * p</t . !3& ! " Km; * * #_a6 A; "! :! # ! $#%&' $#() . 4$ " A & . . + aQ . ' ,S ! " ( K[/() 3$* "D$ g E ,*! . ' K 34 4W - "D$ J;d :! # ! $#%&' $#() ,*! . ' K 63:CH! ": " (L. W L. "! !"! !G * "R "!6 L. W L. t *! "! 3$* ": " (L. W L. ( K) & g E I;"J * !"! A J * "R "!6 L. W L. *! L. ( K) A &W g E ,*! . ' K 63:CH! "!6 KL. W L. *! "! & J;d .! A " * J.! . @; K 63:CH! ": " (L. W Vol. 15, No. 4, Winter 2011 ,23- ( 01 / 0,. . ( )* 0 % . ( / % %' -" % #$ . !" ( +, !" 12% $2 +' &'%" ( )* .(6 5) ( 3 -+ % / +2 / +, !" $ %' -" . 3.5 3 * 2.5 ** 2 1.5 ** 1 0.5 0 2.5 2 * 1.5 ** ** ** ** 1 0.5 0 / .( + , ) & ! "#$% & '() " ( .1 ( ) 6"7 8 ! 19: p<2/25** p<2/23* .1 , 0 ( /) $ ( .(G 5) +2 >2 ' " ! "# $ %# , -&./ - 0$& () * + 6= 0 /2 ? " ( L- 1 22? /1?H %' -" : "# $ %# /) #$ .(%F %EE /%C B ""3 ) '5 / 3 Vol. 15, No. 4, Winter 2011 ,-. (# ')* ( + !" # $%& ' .(% $ ) ! " #$ NAME 120 100 80 * 60 ** 40 ** ** ** 20 0 .3 1 2 - /01 .%&' ( )*+ ,% ! "#$ # ( .3 ( ) 8%! 9 :; 3<= p<4/47** p<4/45* 3.5 3 2.5 * 2 1.5 1 0.5 0 2.5 2 1.5 * 1 0.5 0 D . ( 1 ! , ; ; $C 8 = @ $ / ( A9 ,8 B - ,%8 9 # ( .3 :E; 8%! 9 :; 3<= p<4/47* .3 1 2 Vol. 15, No. 4, Winter 2011 ,.3- ( 12 0 1,/ MA : .,-79 M 1 >! '! L* ! " _N 1 , ) '! # $ , .[^^] !$) (NMDA 1 , )) iG+1 L7! ! " #$% & '! # $ () # *+ ,- NK 801 h NMDA 1 , ) 1 J9$)7+ F + , ! 6#7 .[34] 2#!+ /0$ 1 .$! ,G! h* ([^d] , 1* 1 # $ 7 ! '! 8 * ( * 79* :$;* #$% , ) # * H * # F # EF H "G = # -< .$&! ! " & !! " " 1* 1 ('+ '! M 27G g7B! @ $ .$* ! " & @ " '! * A (> )$? * > )! .,-&* 7& JY ! " $!+ /# ., B* C 1D* 1 ( !! " @ $* G H "G (! ; EF G H "G #$% 2# 1$G * A ! " $!+ > " '! * A 1 2 0 # & > " J9 $ ,$&! I$9! > " , : .,$* ! " & I'7 '! @ " '! * A H "G '- @ (,- '! ! " %# 1 , $* *'7 c! " ( ! " # & 1 )%7 $! K # * .$* ! " & JY @ ! # %# *'7 & 1 , I'7 $F L* #$% & @ '! #,-)7G X 7-G ,--'! 1 L * ., MN JG OP$ * >D .JG $* # H "G # iG+ G7! ,-# ,-! (1,l 7G Z I'7 ,P 1 ) *+ Q* H "G ,; (7&R) : I'7 ,--! '! *$Y* " # (/9G ,G .[TU] JG ,& S ,-! 1,l 7G I'7 ,P 1 -< .,- ! Q H "G ( ! (! P@ .[^T] ,--! '! " # 7! ) (,$* ! " # & I'7 '! * A 2 ( G mG$ ! " $!+ > " & I'7 '! H "G G 7* 9* ( B* C @ n,&* # L # * JG ! + W'! 1 F $0 1 )%7 -0 $! 7"# # .$* #'-* W#+ o9 $G W#+ '! * # JG ! _T D 7@ (JG $* G V @ W H "G 1 @ W Q " # (JG ,* #,-)7G X ,--,$ ; #'-* EF # > , * JM9 1 7'* B* C .,&* ! # 1 2# 1 F JG ! -< .JG ,l$$?$?$) 1 $!$ M& 1 C # JG ! _^ 79#* 7G G $X F (,-&* 7& JY ,& /# 8 * ( 1 F # # ,-&* 7& . ) A G * $! '+ B* C V,S* : ,& p$" ,Z 1 A J 9A 1,l$$$$) 1 $!$ > " JG A .,-&* ,-? '! 2#!+ /0$ .$! ! " # & /f $ " 1,-*q F /# ! # #) * 7Z+ 7 A & > '! \# $ 7" ) A C J ( ! " $!+ *'7 _*+ -< .[T^] .$! .$) JG$X .,-*#! Q /0$ .$! > ,`X . JG 79 $ 1 , ) 1 J9$)7+ #$% ( P@ 6#7 MN : * $B P@ .[^a] ,- '! 2#!+ '! 8 * (77-G,9 /# 7 W#+ )'! !$) (1,l$X $Y '*! : C #$% (MA 1 9! > F + .,& > H " C '! # $ 2* # . 1 ,l$X # N $Y C J#' (M W79G ,P ( G ( ,-! * JG I'7 1,l$X 1 , ) '! ,G! h* (,--! H 1 , ,A # 1 2# 1 F D 7@ H "G 1,P : C '! $Y B* C JG 79 $ 7@ * ,--? '! ($* ! " # & ? $@ ,* ,&HS" * F$ * (,& 7 ) D* $N -< ., JG + 2* ! .,- ) * 1 J@ * , # $Y NMDA 1 , ) 79$)7+ C ( # @ " '! # $ ( + *+ bcY * NMDA , ) J9$)7+ C * C (, ! (MA : eG * F + .[Td] ! " $!+ JG OP$ * >D .,& '! 77-G,9 /# 7 W#+ )'! ,#,& /# & $* /f $ , ( ! " $!+ @ " W#+ ,&HS" # N NMDA 1 , ) : C M# ,G! h* ([^T] JG $@ 1X g $F$! 1 #X * )! >% ,#9 /# 7 ,& 77-G,9 /# 7 $!+ " # '! # $ V H "G Vol. 15, No. 4, Winter 2011 ,-. (# ')* ( + !" # $%& ' 10- Akhondzadeh S, Fallah-Pour H, Afkham Kh, Jamshidi NMDA AH, Khalighi-Cigaroudi F. Comparison of Crocus sativus L. and imiperamine in the treatment of mild to moderate ! "# $ " % &'( ) ( * *!)+ depression: A pilot double-blind randomized trial. BMC ., - ,. Complement Altern Med. 2004;4:12-6. 11- Noorbala AA, Akhondzadeh S, Tahmacebi-Pour N, Jamshidi AH. Hydro-alcoholic extract of Crocus sativus L. versus fluxetine in the treatment of mild to moderate depression: A double blind, randomized pilot trial. J ! "# " - $ " 2 3!* 0 1(" !* &'( Ethnopharmacol. 2005;97:281-4. 12- Hosseinzadeh H, Younesi HM. Antinociceptive and anti- 4! 5 6 $ * 7 , 8 inflammatory effects of Crocus sativus L. stigma and petal extracts in mice. BMC Pharmacol. 2002;2:7-15. 9 & + 9 $ : ;* <= .- 13- Arbabian S, Izadi H, Ghoshooni H, Shams J, Zardooz H, $ " 2 0 * *!)+ NMDA 093!> Kamalinejad M, et al. Water extract of saffron (Crocus sativus) inhibits chronic phase of formaline test in female mice. . ,. ! "# " - Kowsar J. 2009;14:11-8. [Persian] 14- Abe K, Saito H. Effects of saffron extract and its constituent crocin on learning behaviour and long-term 0 9?# @proteins and antiopiates. Peptides. F, Kamalinejad M, et al. Antinociceptive and anti- 1998;19:1603-30. inflammatory effects of Elaeagnus angustifolia fruit extract. J 3- Waldhoer M, Bartlett SE, Whistler JL. Opioid receptors. Ethnopharmacol. 2000;72:287-92. Ann Rev Biochem. 2004;73:953-90. 19- Schmidt M, Betti G, Hensel A. Saffron in phototherapy: 4- Law PY, Loh HH, Wei LN. Insights into the receptor Pharmacology and clinical uses. Wien Med Wochenschr. transcription and signalling: Implications in opioid tolerance 2007;157:315-9. and dependence. Neuropharmacology. 2004;47:300-11. 20- Shams J, Molavi S, Marjani S, Kamalinejad M, Zardooz 5- Simmons DL, Botting RM, Hla T. Cyclooxygenase H, Sahraei H, et al. The aqueous extract of Crocus sativus isozymes: The biology of prostaglandin synthesis and stigma reduces morphine tolerance. Physiol Pharmacol. inhibition. Pharmacol Rev. 2004;56(3):387-437. 2009;13(2):170-8. [Persian] 6- Payan DG. Nonsteroidal anti-inflammatory drugs, 21- Shibata M, Ohkubo T, Takahashi H, Inoki R. Modified nonopioid analgesics, drugs used in gout. In: Katzung BG, formalin test: Characteristic biphasic paine response. Pain. editor. Basic and clinical pharmacology. USA: Appleton and 1989;38:347-52. Lange; 1992. 22- Popik P, Kozela E, Danysz W. Clinically available NMDA 7- Huang KCH. The pharmacology of Chinese herb. 2nd ed. receptor antagonist’s memantine and dextromethorphan USA: CRC Press; 1999. reverse existing tolerance to the antinociceptive effects of 8- Zargari A. Medical plants. Tehran: Tehran University Press; morphine in mice. Pharmacology. 2000;361:425-32. 1990. [Persian] 23- Lechtenberg M, Schepmann D, Niehues M, Hellenbrand 9- Agha-Hosseini M, Kashani L, Aleyaseen A, Ghoreishi A, N, Wunsch B, Hensel A. Quality and functionality of saffron: Rahmanpour H, Zarrinara AR, et al. Crocus sativus L. Quality control, species assortment and affinity of extract and (saffron) in the treatment of premenstrual syndrome: A double- isolated saffron compounds to NMDA and σ1 (Sigma-1) blind, randomised and placebo-controlled trial. BJOG. receptors. Planta Med. 2008;74:764-72. 2008;115(4):515-9. Vol. 15, No. 4, Winter 2011 ,23- ( 01 / 0,.