PJ BS Bossxiczs so Pakistan Journal of Biological Sciences ANSI ez n Network for Scientific Information 308 Lasani Towns Sargodha Road, Faisalabad - PakistanPakistan Joumal of Biological Seienees 10(15): 2583-2586, 2007 ISSN 1028-8880 (© 2007 Asian Network for Seientifie Information Interaction Between Curcumin and Opioid System in the Formalin Test of Rats “Hossein Tajik, “Esmacal Tamaddonfard and “Nasrin Hamzch-Gooshehi "Department of Food Hygiene, “Department of Physiology, College of Veterinary Medicine, Urmia University, Urmia, Iran ‘Abstract Tn this sty, the effeet of cursumnin onthe formalin-induced pain was investigated in rata, Tnieraction between eueunin and opioid system using morphine and naloxone was abo examined. A biphasie pain resporse Was induced after intraplantr injection of formalin (30 yi, 196). Cureumin, moqphin and naloxone had no effect on the early phase of pain. Late phase of pain was suppressed by curcumin atthe doses of 100 and 200:mg kg” body weigh. Morphine (1 mg kg" EW) reduced, whereas naloxone (1 mg kg BW) didnot affect the late phase of pain Curreumin did not influence the morphine-induced antinociception, but reversed the cfeet of naloxone om pain, Present findings indiate thar curcumin may produee antinoeiveption by aetivatice ‘of both epicid and non opioid mechanisms of pain. Key words: Curcumin, morphine, naloxone, formalin-induced pain rats INTRODUCTION ‘Curcumin is the prominent yellow pigment in mmeric (Curcwma longa), a widely used spice and food ‘colouring agent with anti-inflarumatory, anti-oxidant, lanli-angiogenic, anti-bacterial ane anti-cancer properties (Gammon and Wabl, 1991; Mabeshnvar ea, 2006). On the antinociceptive effect of curcumin, it was reported that ‘cucumin produced antinociception using tail immersion ‘and hot plate assays of pain ina diabetic mouse model of| ‘neuropathic pain (Shurma ef af, 2006). JCICM-6 is an ‘extract of an anti-arthrite herbal formula and Cureuma fonga is one of its components, It was found that JICICM.6 produced antinociception in oth tail ic of rats ‘and writhing reflex of mice (Zhou et af, 2006), ‘The formalin est isan important animal model in the study of acute long-lasting inflammatory pain (Pu et al, 2001) In this model, inraplantar injection of formalin into ‘2 hindpaw clicts a biphasic pattem of pais-related behaviours, an early short-lasting neurogenic phase followed by a second and more sustained inflammatory phase (Tjolten et a, 1992) 1 was reported dhat $1627, an inhibitor of nuclear factor kappaB, reduced the late phase of formalin pain in rats (Tegeder ef af, 2008), It was found that cureumin inhibited the activation of nuclear factor kappa? using the Panomies FkappaB Receptor Stable Cell Line (Weber etal, 2006} To date, the effect of cureumin on the formali induced pain was not reported. This the preseat study ‘was designed to investigate the effect of curcumin om the formalin-induced pain Cwreumin effect onthe ‘endogencus analgesic opicic system was also examined using morphine (an opioid agonist) and naloxone Can opioid antagonist), MATERIALS AND METHODS, Animal: Healthy adhlt mae albino wistar tats (220-250 g) ‘obtained from the Animal Care and Use Center of Umi ‘University, Rats were maintained in polypropylene cages in four groupe with food and water available df [itum, ‘with controlled ambient temperature (20-23) and under & 12h light-dark eyele (lights on 0700 blights off 19.00), [Eight rats were used in each group. The experimental ‘protocol sas approved by the Laboratory of Animal Care and Use Center of Urmia University [Drugs and treatments: Drugs used in the present study ‘were curcumin (Merck, Damastait, Germany), morphine sulphate and naloxone hydrochloride (Temad, Tehran, Tron). Curcumin is insoluble in water, but is soluble im ‘ethanol, alkalis, Ketone, acetic acid and chloroform ‘Araujo and Leon, 2001). Therelore, eureumin suspension ‘was prepared in 5% ethanol solution, Cureumin suspension was freshly prepared and administered orally at the doses of 5, 50, 100 and 200 mg kg” body weight 445 min before fermalin injection Curcumin administration ‘was made in a constant volume of 1 mL“ for each rat ‘vera period of 3-5 min. Morphine and naloxone were disolved in normal saline and were injected intraperitonealy aad subcutanecwsly 30 min and 1b ‘Corresponding Author: Dr Hosssin Tajik, Department of Food Hygiene, College of Veterinary Medicine, Unmia University, Unmia, an Tel:81 441 2770808 Fae 81 441 2771926 2583Pale. J. Biol. Soi, 10 (19): 2583-2586, 2007 before formalin injection, respectively, In combined treatments, morphine and naloxone were injected 15:min after or before eurcumin administration, sespectively ‘Formalin test: Each rit was placed inside a plexiglass ‘observation chamber for an acclimation period of 30:min, At the end of this period, drug treatment was performed according to the time schedule for each treatment and then 50 jL of 1% formalin was subedraneously injected into the plantar region of the right hindpaw using a 29- gauge injection needle, Control group was intraplantarly injected With normal saline, Immediately after fosmalin and ‘normal saline injections, the time spent licking and biting the injected paw was reoorded in 5 min intervals for 1 I (Token ef af, 1992). In present study, data collected between D and 5 min post-formalin injection represented phase one (early phase) and data collected between 116-1S min ater injection of formalin represented phase to (ate phase) ‘Statistical analysis: Data were expressed as means+SEM, Differences among treated groups were statistically ‘evaluated wing the onesway analysis of variance (ANOVA) followed by Duncan's test, Differences were ‘considered significant at p05. RESULTS Figure 1 shows the effect of curcumin on the formalin-induced pain response, Invaplantar injection of ‘mimo! 0, 59) + Nowa min.) ‘mmol (0, 5%) + Foran ip 1%) ‘caren 25mg hsp) + Ferman ip, 1%) ‘3 cazeoin(S0mg ha” 30)-+Fermaln Op 96) ‘arcmin (100mg es") +Fomalin (ip 1%) ‘Boreumin 200mg 7.0) + Fonsi Gp 1) Dream of king sd beg he aed po) wy pula)" La phase (16-45) Fig.l: Effet of curcumin on the formalin-induced pain in rats, Values are mean!SEM, *p0.05 vs. olher groups in the Inte phase, por per oral, ipl intraplantar 1m haa (90, 599 + Normale) 1 Cuvenin 100mg pn.) ~ Forman 1%) 1 Neca agg 12) Forma, 1%) 1 Neboune (mpg, p0) Fema 1%) 1 Cacunin (100mg a”. p.0)+ Morphin (Gragig stp) Fema p19) 1G Nadowne gg", 46) + Cai (Goosgig 9.0) Formalin Op) Durtnaf king and nae nocd ow) ‘eeseseseseg Belyphse(0Smia) "Lape (1645 ml) ‘Fig. 2: Effect of eweumin on the morphine and naloxone= induced pain changes in rats, Values are meantSEM, "p=0.05 va. ethanol+formalin and nalosone+formatin groups in the late phase, pc: per oral, ip intraplantar, sc: subcutaneous formalin produced a marked biphasie pain (early phase: 59.5474 5; Inte phase: 222.7184 s). Curcumin at the doses of 25, 0, 100 and 200 mg kg” body weight had no effect on the early phase of pain. Late phase of pain was ‘not alfeced by curcumin at the doses of 25 and 50mg kg body weight, whereas curcumin at the dose ‘of 100 and 200 mg kg” body weight significantly ‘suppressed the late phase of pain (p<0.05). No significant difference was observed between the effects of 100 and 200 mg ke body weight of curcumin, Figure 2 shows tho effects of morphine, naloxone ‘alone and with curcumin on the formalin-indweed pain, Early phase of pain was not affected with alone application of curcumin, merphine and naloxone. Pesta pre-curoumin treatments with morphine and) naexone, teapeetively, did not change this phase, too. Curcumin (100 mg kg! BIW) and morphine (Img kg! BW) significantly reduced the late phase of pain (p-0.05), No significant diferenoe was observed between the effects of ‘curcumin and morphine, Naloxone (1 mg kg~ BW) did not ‘change the late phase of pain. The effect of moqphine on the Inte phase was not influenced by cureumin pee- treatment, Posttreatment with curcumin significantly decreased the effect of naloxone on pain (90.05), DISCUSSION In the present study, it was found that curcumin suppressed inflammatory pain. The lack of effect of ‘cureumin, morphine and naloxone on the early phase of, 2584Pale. J. Biol. Soi, 10 (19): 2583-2586, 2007 pin observed in the present study, may be rented to the fact thatthe early phase of formalin pain is a neurogenic pain and produces due fo direct activation of C-fiber nociceptors (Tjolsen ef af, 1992) Ik was reposted dat nti-inflammatory drugs did not affect the first phase of formalin pain even when a very law formalin ‘concentration used (Rosland ef al, 1990), Moreover, it ‘was found thatthe potency of mogphine inte formalin test affected by both the way of administration and formalin concentration (Servostianava et af, 2003). It this study, curcumin at the doses of 25 and 50:mg lg"! body weight did not influence pain. It ‘has been reported that curcumin is pocely absorbed inthe intestinal waet. Oral doses are largely exereted in facoes ‘and only twace amounts appears in the blood (Ravindranath and Chandrasethara, 1980; Ammon and ‘Wahl, 1991). ‘The antinceiceptive effect of curcumin on the inflammatory pain observed inthis study may be related tots ant-inflammatery property It hs been reported that ‘eurenmin has abilily to inhibit the activation of inflammatory mediators, such as eyclocxygenase 2, lipooxygenase, inducible nitric oxide synthase and ‘nuclear factor kappa Benamark, 2006), On the other ‘hand, the contsibution of histamine, serotonin, bradykinin, cytokines, sympathetic amines, lipoxygenase, cyclooxygenme one and two, inducible nitric oxide synthase t0 one ot both phases of formalineinduced pain is firmly esublished (Tjolsen et af, 1992. Doak and Sawynok, 199% Doursout ef al, 200 (Chichorro et af, 2004), In this study, cweumin with no effect on the ‘morphine-snduced | antinociception, atferniated the naloxone effect, This indicates that curcumin produces antinociception ly activation of both opioid and non ‘picid pain mediated systems. The sensation of pa is Well know to be modified by endogenous opioid and nem ‘opioid systems. Morphine (an opioid) agonist) and naloxone (an opioid antagonist) are used to explore the involvement of endogenous analgesic aystoms activated bby novel analgesics (Yoshimatsu and Furue, 2006, Anunthan, 2006) On the other ban, in bilateral Olfactory Bulbuectomy (OB) model of depression in rats, it we reported that curcumin changed the moncaminergic neurotransmitter levels such a Shydroxstriptamine, rnoradrenalin and dopamine and their metabolites in the hippocampus and frontal cortex of brain (Xu et af, 2005), ‘Those neurotransmitters comprise the important part of ‘non opioid endogenous pain modulating system (Yoshimatsu and Furie, 2006) Finally, it seems that several mechanisms. are involved in the analgesic effect of curcumin in the formalin test Further studies are needed to identify ‘mechanisms, REFERENCES Ammon, HLP. and MA, Wahl, 1991. Pharmacology of CCurcuma tonga. Planta Med, $7: 1-7. 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