Investigation of antidepressant and anxiolytic activity of curcumin given alone and in combination with amitriptyline in rats
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Pankti Patel et.al Indian Journal of Research in Pharmacy and Biotechnology ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online)
Investigation of antidepressant and anxiolytic activity of curcumin given alone and
in combination with amitriptyline in rats
Pankti Patel*, Kashmira J Gohil, Samaresh Pal Roy, Nikunj Patel
Department of Pharmacology, Shree Dhanvantary Pharmacy College, Kim- 394110, Surat, Gujarat, India.
*Corresponding author: Email: panktip691@yahoo.co.in; Ph.: +91-9714000750
ABSTRACT
The objective of the present study was to evaluate and investigate the antidepressant and
anxiolytic activity of curcumin given alone and in combination with amitriptyline in rats. The
investigation of antidepressant activity was carried out using Despair Swim Test (DST) and Tail
Suspension Test (TST) and for anxiolytic activity Open Field Test (OFT) and Elevated Plus Maze Test
(EPM) at 14th day. Wistar albino male rats were divided into six groups consisting six animals each.
Control Group I received 2% Gum Acacia (Vehicle control), standard groups Group II and Group III
recieved Amitriptyline (AMI) 10 mg/kg and Curcumin (CUR) 70 mg/kg respectively and combination
groups Group IV, V and VI received CUR 35 mg/kg + AMI 5 mg/kg, CUR 70 mg/kg + AMI 5 mg/kg and
CUR 70 mg/kg + AMI 10 mg/kg respectively for 14 days. Duration of immobility and locomotion,
number of rearings, number of squares crossed, and time, spend and number of entries in open and closed
arms was evaluated for antidepressant and anxiolytic activity respectively. Combination therapy of
curcumin and amitriptyline showed significant decrease in duration of immobility in DST and TST,
increase in locomotion, number of rearings and squares crossed in OFT and increase in number of open
arm entries and time spend in open arm in EPM as compared to control and standard group of curcumin
and amitriptyline alone. The results revealed that curcumin when given in combination with amitriptyline
showed synergistic antidepressant and anxiolytic activity, which may reduce side effects of amitriptyline
in chronic depressive patient.
Keywords: Curcumin, antidepressant, amitriptyline, anxiolytic
INTRODUCTION neurotransmitters such as serotonin, noradrenaline and
Depression and anxiety are heterogeneous dopamine (Xu et al., 2005). Curcumin promotes the
mood disorder that has been classified and treated in hippocampal neurogenesis (Xu, 2007; Xu, 2006;
various ways. Across the globe, depression and Wang, 2008). Amitriptyline, Tricyclic Antidepressant
anxiety are major cause of disability. A close (TCA), used as antidepressant acts by inhibiting the
relationship between anxiety and depression has been reuptake of serotonin and noradrenaline to large extent
acknowledged since ancient times. Thus if anxiety is and dopamine to a lesser extent (Rang, 2007). It has
an integral aspect of depression using the treatment of many side effects due to chronic administration which
depression, anxiety may also be treated (Hranov, overcomes its benefit. Dose reduction of amitriptyline
2007). Tricyclic antidepressants are effective in when it is given in combination with curcumin may
treating both depression and anxiety (Rickels and decrease the side effects of amitriptyline and may
Schweizer, 1993). prove to be cost effective, thus this drug-herb
Although a number of synthetic drugs are interaction may be beneficial and may produce
being used as standard treatment for depression and synergistic effect of both curcumin and amitriptyline.
anxiety, they have adverse effects that limit the MATERIALS AND METHODS
therapeutic treatment. Traditionally herbs are used for Animals: Wistar albino rats weighing 150-250 gm
depression and anxiety which may offer advantage in were procured from Zydus Cadila Healthcare Ltd. as a
terms of safety and tolerability, possibly by gift for research endeavour. They were housed in
improvement in patient compliance (Richelson, 1994). group of six and were fed on standard pellet diet and
Curcuma longa (Turmeric) is the most widely water ad libitum and maintained in environmental
used herb as an Indian spice with numerous controlled room at 25+ 3 °C and 50 + 20 % humidity
pharmacological activities with antidepressant with 12h light/dark cycle. The experimental protocols
activity. Curcumin as it has high safety profile were approved by the Institutional Animal Ethical
(Ahmad, 2011). Curcumin acts as an antidepressant by Committee (IAEC) and experiments were conducted
variety of mechanisms. Curcumin also inhibits the according to the guidelines of CPCSEA (Committee
monoamine oxidase enzyme which increases the level for the Purpose of Control and Supervision of
of neurotransmitters in the synaptic cleft of neurons Experiments on Animals).
(Kulkarni, 2008). Curcumin modulates the level of
IJRPB 2(3) www.ijrpb.com May-June 2014 Page 1173Pankti Patel et.al Indian Journal of Research in Pharmacy and Biotechnology ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online)
Curcumin: Curcumin was obtained as a free gift evaluating potential antidepressants. Rats were
sample from Sami Labs Pvt. Ltd. for the project. treated with the test compounds or the vehicle by p.o.
Curcumin suspension was made in 2% gum acacia for administration 60 min prior to testing. For the test, the
administration. rats were suspended on the edge of a shelf 58 cm
Drug and chemicals: Amitriptyline hydrochloride above a table top by adhesive tape placed
was received as a generous gift sample from IPCA approximately 1 cm from the tip of the tail. The
Laboratories Ltd. Amitriptyline hydrochloride was duration of immobility was recorded for period of 6
dissolved in distilled water for administration. Gum min. Rats were considered immobile when they hang
Acacia, Butanol, Hydrochloric acid, Heptane, Sodium passively and completely motionless.
acetate, Sodium Hydroxide, Iodine, Glacial Acetic Elevated Plus maze (EPM): Based on Montgomery
acid, Sodium sulfite, O-phthaldialdehyde reagent, (1958), Pellow et al. (1985) and Corbett et al. (1991),
Noradrenaline, Serotonin and Dopamine were elevated plus-maze apparatus consists of two open
procured from Shree Dhanvantary Pharmacy College, arms, 50×10×40 cm, and two enclosed arms,
Kim, Surat. All the chemicals and reagents used were 50×10×40 cm, with an open roof, arranged so that the
analytical grade. two open arms are opposite to each other. The maze is
Dosage administration: Animals were randomly elevated to a height of 50 cm. The Curcumin or
divided in to six groups consisting of 06 animals per vehicle was administered for 4 weeks once daily p.o.
group as mentioned below. All the test solutions were and the last dose was given on the 28th day, 60 min
freshly prepared daily and administered in animals prior to experiment. The rat was placed in the center
orally for 14 days by p.o. route. of the maze, facing one of the enclosed arms. During a
Groups Treatment 5 min test period, the number of entries into open and
Group I Vehicle (Gum acacia 2%) enclosed arms, time spent in the open and enclosed
Group II Amitriptyline (AMI) 10 mg/kg arms, total number of arm entries and percentage
Group III Curcumin (CUR) 70 mg/kg preference to open/enclosed arm was observed. An
Group V Curcumin (CUR) 35 mg/kg and arm entry was defined as the entry of all four paws
Amitriptyline (AMI) 5 mg/kg into the arm.
Group V Curcumin (CUR) 70 mg/kg and Open field test (OFT): Measure of movements of rats
Amitriptyline (AMI) 5 mg/kg in a cage (“open field”) has been used by Dews
Group VI Curcumin (CUR) 70 mg/kg and (1953), Saelens et al. (1968) and Nakatsu and Owen,
Amitriptyline (AMI) 10 mg/kg (1980). Movements of rats were observed in a square
Despair swim test (DST): The study was carried out open field arena (68×68×45 cm) for various
on rat according to the method of Porsolt et al.(1977, parameters.Treatments were given orally 60 minutes
1978). When the rats were placed in the vertical open before test. The rats were observed for 5 min in a
cylinder (height: 40 cm; diameter: 18 cm) containing dark, ventilated, sound-attenuating box and following
15 cm of water maintained at 25 °C for the first time variables were evaluated: Locomotion, number of
initially highly active, vigorously swimming in rearings, number of field segments entered and
circles, trying to climb the wall or diving to the number of fecal boluses.
bottom. After 5–6 min, the rats remained immobile for Statistical analysis: The data obtained were presented
approximately 80% of the time. After 15 min in the as mean + S.E.M (n=6) and ***Pankti Patel et.al Indian Journal of Research in Pharmacy and Biotechnology ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online) decreased in combination group receiving curcumin The antidepressant activity of curcumin and 70 mg/kg and amitriptyline 5 mg/kg (74.16 + 0.74) as amitriptyline was evaluated by Despair Swim Test compared to control group (157.5 + 0.42) and (DST) and Tail Suspension Test (TST). The standard group receiving curcumin 70 mg/kg alone combination of curcumin and amitriptyline showed (97.83 + 1.31). significant decrease in duration of immobility Effect of Curcumin alone and in combination with comparable to that of standard amitriptyline and Amitriptyline on Immobility time in rats in Tail curcumin after 14th day of chronic treatment. DST suspension test (TST): On 14th day of treatment in based on the observation that animals in an TST, the immobility time was found to decrease in inescapable cylinder filled with water and in TST, combination group as compared to vehicle treated and when suspended by their tails using adhesive tape to standard groups. (Table 2; Figure 2) There was a table top, initially are engaged in escape-like significant (***p< 0.001) decrease in immobility time behaviour followed by developing immobile posture. in combination group receiving curcumin 70 mg/kg Immobility is interpreted as a passive stress-coping and amitriptyline 5 mg/kg (53.67 + 1.77) as compared strategy or depression-like behavior (behavioral to control group (119.33 + 0.42) and standard group despair). After antidepressant administration, the receiving curcumin 70 mg/kg alone (90.83 + 0.74). animals actively perform escape-directed behaviors Effects of Curcumin alone and in combination with with longer duration than animals with control amitriptyline on % preferences towards open arm treatment (Demouliere et al., 2005). Earlier study in rats in Elevated Plus Maze test (EPM): In reported that NMDA receptor involved in Elevated Plus Maze Test, combination group antidepressant activity of curcumin in DST and TST receiving curcumin 70 mg/kg and amitriptyline 5 model (Lin et al., 2013). A significant decrease in mg/kg showed no significant results in number of immobility was observed in a combination of both entries but showed significant (***p< 0.001) results in DST and TST and it may be due to ability of curcumin time spent in open and closed arm than that of and amitriptyline to modulate the brain standard curcumin (70 mg/kg) alone and amitriptyline neurotransmitter level. (5 mg/kg) on 14th day but no significant results were Anxiolytic activity was measured by Open observed in number of entries in open and closed arm Field Test (OFT) and Elevated plus maze test (EPM). (Table 3; Figure 3). In Open Field Test (OFT), rats when treated with Effect of Curcumin alone and in combination with combination of curcumin and amitriptyline produces Amitriptyline in rats in Open Field test (OFT): In significant increase in locomotion, number of rearings OFT, Amitriptyline (10 mg/kg) treated rats showed and number of field segments entered. Higher levels significant increase in number of rearing, number of of anxiety should mainly lead to decreases in the ratio squares crossed and locomotion during 5 min interval ‘number of squares visited in centre/number of as compared to control group (table 4). Combination squares visited on periphery. In experiments involving groups receiving curcumin (70 mg/kg) and rodents, observers do not measure the effects of amitriptyline (5 mg/kg) produced significant treatments on exploration, but the reaction to a (***p
Pankti Patel et.al Indian Journal of Research in Pharmacy and Biotechnology ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online)
of open and closed arms whereas significant results antidepressant by variety of mechanisms. Curcumin is
were observed in time spent in open and closed entries known to play a role in increasing hippocampal
and % preference to open arm. This model is used for BDNF levels (Hurley et al., 2013), stimulating
evaluation of anxiolytic drugs as exposure of animal neurogenesis (Xu et al., 2005, 2006) (Wang et al.,
to external and internal stimuli are assumed to be 2008) and involvement of glutamate receptor (Lin et
capable of producing anxiety (Bourin et al., 2007). al., 2013). Thus the synergistic effect of curcumin and
Based on the results obtained, the possible amitriptyline as an antidepressant and anxiolytic
mechanism of curcumin may be the modulation of the observed may be because of combination of all this
level of brain neurotransmitter, serotonin, mechanisms.
noradrenaline and dopamine. Curcumin works as an
Table.1.Effects of Curcumin alone and in combination Table.2. Effect of Curcumin alone and in combination
with Amitriptyline on immobility time in rats in Despair with Amitriptyline on Immobility time in rats in Tail
Swim Test (DST) Suspension Test (TST)
Groups Immobility time (sec) Groups Immobility time (sec)
14th day 14th day
Group I:Control 157.5 + 0.42 Group I:Control 119.33+0.42
Group II:AMI 10 mg/kg 66.66 + 0.79*** Group II:AMI 10 mg/kg 47.83+0.50***
Group III:CUR 70 mg/kg 97.83 + 1.31*** Group III:CUR 70 mg/kg 90.83+0.74***
Group IV:CUR 35 mg/kg+ 86.83 + 0.81*** Group IV:CUR 35 mg/kg 82.83+0.79***
AMI 5 mg/kg and AMI 5 mg/kg
Group V:CUR 70 mg/kg+ 74.16 + 0.74*** Group V:CUR 70 mg/kg 53.67+1.77***
AMI 5 mg/kg and AMI 5 mg/kg
Group VI:CUR 70 mg/kg+ 61.33 + 0.46*** Group VI:CUR 70 mg/kg 34.16+0.74***
AMI 10 mg/kg and AMI 10 mg/kg
Values represent the mean+ S.E.M.(n=6). ***pPankti Patel et.al Indian Journal of Research in Pharmacy and Biotechnology ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online)
180 140
Immobility time (sec)
160 120
140
Immobility time (Sec)
100
120 ***
*** 80 ***
100
***
80 *** 60 ***
*** *** ***
60 40
40
***
20
20
0 0
Figure 1: Immobility time (Sec) in rats in Despair Figure 2: Immobility time (Sec) in rats in Tail
Swim Test. Values represent the mean+ Suspension Test. Values represent the mean+
S.E.M.(n=6). ***pPankti Patel et.al Indian Journal of Research in Pharmacy and Biotechnology ISSN: 2321-5674(Print) ISSN: 2320 – 3471(Online)
CONCLUSION Lin et al., NMDA GluN2B receptors involved in the
From the above results, it can be concluded antidepressant effects of curcumin in the forced swim test,
that the combination therapy of Curcumin and Progress in Neuro. Psychopharmacol. Bio. Psychiat, 40,
Amitriptyline showed a significant result as compare 2013, 12–17.
to the individual administration of standard Montgomery KC, The relation between fear induced by
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significant decrease in immobility time in Despair Nakatsu K and Owen JA, A microprocessor-based animal
Swim test and Tail Suspension test which confirms its monitoring system, J. Pharmacol. Meth, 3, 1980, 71–82.
antidepressant activity. From Open field test and
Pellow S, Chopin P, File SE, Briley M, Validation of
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groups which confirms its Anxiolytic activity. Hence
the synergism of curcumin and amitriptyline may be Porsolt RD, Anton G, Blavet N, Jalfre M, Behavioural
despair in rats: a new model sensitive to antidepressive
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This work was supported by a grant from Gujarat Int. Pharmacodyn, 229, 1977, 327–336.
Council on Science and Technology Rang HP, Dale MM, Ritter JM and Flower RJ, Rang and
(GUJCOST/MRP/12-13/56/1336). Dale’s Pharmacology: Antidepressant drugs, (6th Edn)
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