|Year : 2017 | Volume
| Issue : 4 | Page : 215-219
Anti-anxiety and anti-depressant like effects of murraya koenigii in experimental models of anxiety and depression
Snigdha Sharma, Shailendra Handu, Ashok Kumar Dubey, Prashant Sharma, Pramod Mediratta, Qazi Mushtaq Ahmed
Department of Pharmacology, School of Medical Sciences and Research, Sharda University, Greater Noida, Uttar Pradesh, India
|Date of Web Publication||28-Nov-2017|
Ashok Kumar Dubey
Department of Pharmacology, School of Medical Sciences and Research, Sharda University, Greater Noida, Uttar Pradesh
Source of Support: None, Conflict of Interest: None
Background: Presence of free radical scavenging activity in Murraya koenigii, commonly known as Curry leaves, has been shown in previous studies. Oxidative stress plays an important role in the development of various neurobehavioral disorders including anxiety and depression. Aim: The present study aimed to evaluate the effects of Murraya koenigii in animal models of depression and anxiety. Materials and Methods: The effect of incremental doses of Murraya koenigii aqueous leaf extract was evaluated on spontaneous motor activity (SMA), open arm incursions in elevated plus maze, and despair behaviour in forced swim (FST) and tail suspension (TST) tests as compared to control groups in Swiss albino mice. Results: Murraya koenigii 300 mg/kg, p.o. (MK300) and 400 mg/kg, p.o. (MK400) reduced the SMA count from 754 ± 64.9 to 540 ± 29 and 295 ± 34 respectively, which was statistically significant. MK300 and MK400 reduced significantly the open arm count from 29 ± 8.6 to 16 ± 7 and 10 ± 3.9, respectively. On FST, MK400 reduced the duration of immobility from 145.5 ± 29 to 91 ± 17.3, which was statistically significant. On TST, MK produced a dose-dependent decrease in the duration of immobility; however, it was statistically significant only with MK400. Conclusion: Murraya koenigii aqueous leaf extract reduced the despair behavior in experimental animal models, suggesting an anti-depressant like activity. Murraya koenigii extract also reduced spontaneous locomotor activity in a dose-dependent manner suggesting a sedative and/or anxiolytic effect though there wasn't any anxiolytic effect in the elevated plus maze test.
Keywords: Anxiety, depression, Murraya koenigii
|How to cite this article:|
Sharma S, Handu S, Dubey AK, Sharma P, Mediratta P, Ahmed QM. Anti-anxiety and anti-depressant like effects of murraya koenigii in experimental models of anxiety and depression. Ancient Sci Life 2017;36:215-9
|How to cite this URL:|
Sharma S, Handu S, Dubey AK, Sharma P, Mediratta P, Ahmed QM. Anti-anxiety and anti-depressant like effects of murraya koenigii in experimental models of anxiety and depression. Ancient Sci Life [serial online] 2017 [cited 2019 Aug 17];36:215-9. Available from: http://www.ancientscienceoflife.org/text.asp?2017/36/4/215/219368
| Introduction|| |
Anxiety disorders are the most prevalent mental disorders affecting up to 33 percent of the population during their lifetime. Anxiety disorders commonly coexist with other mood disorders such as depression. Depression is also very common in the general population with a lifetime prevalence of 8 to 12 percent. Pharmacotherapy of depression and anxiety is often unsatisfactory with more than one third of the depressed patients not responding properly to the currently available drugs. Even in those who respond, the cost and the associated adverse effects of prolonged therapy with the current medications, have been important limiting factors for proper compliance and effective management.
Murraya koenigii, also known as curry leaf plant, is a tropical to sub-tropical plant belonging to family Rutaceae. Murraya koenigii is found almost everywhere in the Indian subcontinent except at the higher levels of the Himalayas. The bitter aromatic Murraya koenigii leaves are commonly used as a spice and a domestic remedy for many disorders. The aqueous extract of the leaves of the plant have been shown to possess antioxidant activity in previous studies., The studies have established the presence of free radical scavenging activity in Murraya koenigii highlighting its potential to be used for decreasing oxidative stress.
Oxidative stress plays an important role in the development of various neurobehavioral disorders including anxiety and depression. Studies on knockout and over-expression of antioxidant enzymes have shown a relationship between anxiety-like behavior and oxidative stress. A linear relationship has been established between peripheral blood oxidative stress markers and anxiety-like behavior. Oxidative stress has also been shown to be linked with the pathogenesis of depressive disorder. Patients with depression have decreased anti-oxidant defenses and more oxidative DNA damage as compared to non-depressed individuals and studies have found significantly increased serum levels of oxidative stress biomarkers in depressed patients in the acute phase as compared to those of healthy subjects.
Considering the role of oxidative stress in neurobehavioral disorders, a compound with antioxidant activity may be effective in the management of depression and anxiety. Not much research has been done to evaluate the role of Murraya koenigii in depression and anxiety, though its antioxidant activities have been shown in previous studies. Hence, this study aimed to evaluate the effects of Murraya koenigii in the animal models of depression and anxiety.
| Materials and Methods|| |
The present experimental study on animal models of depression and anxiety was conducted as part of the ICMR Short Term Studentship program. Permission was obtained from the Institutional Animal Ethics Committee (IAEC) of the institute. The IAEC is constituted by and registered with CPCSEA, Department of Animal Welfare, Ministry of Environment. None of the tests conducted was of invasive nature and full care of the animals was taken as per “CPCSEA Guidelines” for laboratory animal facilities.
Preparation of plant extract
The leaves of Murraya koenigii were collected locally and authenticated by an authorized center. Aqueous extract was prepared from the powder obtained upon finely grinding the shade dried leaves. The extract was concentrated by evaporation, lyophilized and refrigerated for further use in the study.
Swiss albino mice of either sex weighing between 25 to 30 g were taken for the study. The animals were housed in the central animal house of the institution, under standard laboratory conditions and were kept on pellet diet and water ad libitum. Prior to the day of experiment, animals were kept on fast overnight, though water was given ad libitum. Animals were maintained on 12:12 light- dark cycle, temperature 22 ± 2°C and 30-70% humidity. The animals were acclimatized to laboratory conditions prior to experimentation.
Sample size and the experimental groups
A total of 60 mice of male sex were subjected to the study.
The tests for depression were done in five different groups (n = 6/group) of mice. The vehicle, standard drug and the three test groups were orally administered distilled water (10 ml/kg), fluoxetine (25 mg/kg), and incremental doses of 200, 300 and 400 mg of Murraya koenigii extract respectively, one hour prior to the experiments.
Similarly the tests for anxiety were done in five different groups (n = 6/group) of mice. The vehicle, standard drug and the three test groups were orally administered distilled water (10 ml/kg), diazepam (5mg/kg), and incremental doses of 200, 300 and 400 mg of Murraya koenigii extract respectively one hour prior to the experiments.
There were the following groups (n = 6) to study the effects of Murraya koenigii on anxiety and depression models in mice. Male mice only were used in both the control and drug treated group. The experimental test was performed 1 hour after oral (p.o) administration of vehicle/drug.
- Group 1 – Control: Vehicle of Murraya koenigii, (Distilled water) (10ml/kg, p.o.) – [DW]
- Group 2 – Standard drug: Fluoxetine (20 mg/kg, p.o) for anti-depressant activity or Diazepam 5 mg/kg, p.o for anti-anxiety activity
- Group 3 – Murraya koenigii (200 mg/kg, p.o.) - [MK 200]
- Group 4 – Murraya koenigii (300 mg/kg, p.o.) - [MK 300]
- Group 5 – Murraya koenigii (400 mg/kg, p.o.) - [MK 400].
Experimental models for depression
Forced swim test
Each animal was placed inside a vertical plexiglass cylinder (40 cm in height and 18 cm in diameter), containing water up to a height of 15 cm maintained at 24 – 25°C. After the initial two to three minutes of vigorous struggling activity, an animal usually assumes a typical immobile posture. A mouse was considered to be immobile when it remained floating in the water without trying to struggle, making only minimum movements of its limbs, necessary to keep its head above water. During the test, each animal was placed in the water for 6 min. The duration of immobility was recorded during the last four minutes of the forced swim test. After removing the mice from water they were allowed to dry for 15 min. and then replaced back into the cage. An antidepressant effect was indicated when there was decrease in the duration of immobility.
The tail suspension test
In this test, each mouse was suspended from the edge of a 58 cm high table top with the help of an adhesive tape placed approximately 1 cm from the tip of the tail. The duration of immobility was recorded for a period of 5 minutes. Mouse was considered immobile when it hung passively and remained completely motionless, except for the respiratory movements, for at least 1 min. The duration of immobility in the different groups was compared. A decreased duration of immobility is indicative of antidepressant effect.
Experimental models for anxiety
Elevated plus maze method
The apparatus consists of two open arms, two enclosed arms and a central platform, arranged in such a way that the two arms of each type will be opposite each other. Each animal was placed at the center of the maze facing one of the open arms. During the 5 min test period, the number of open and enclosed arm entries plus the time spent in the open and enclosed arms was recorded digitally. Entry into an arm was defined as the point when the animal places all the four paws on the arm. An increase in the proportion of time spent and the number of entries in the open arm is indicative of anti-anxiety effect.
Measurement of spontaneous locomotor activity
Photoactometer is a chamber that was used to observe locomotor activity of the mice. Animals were placed in a photoactometer which has continuous beams of lights, criss-crossing the chamber and falling on corresponding photoelectric cells. The light beams would get interrupted when the mouse crosses them and these interruptions were recorded for a period of 10 min. Locomotor activity was expressed in terms of total photo beam interruptions.
Data was collected, compiled and analyzed using SPSS software. The data was presented as Mean ± SEM. The data was subjected to appropriate test of significance i.e., Analysis of Variance followed by post-hoc Tukey's Multiple Comparison test. A P value of < 0.05 was considered as significant.
| Results|| |
Diazepam 5 mg/kg reduced the SMA count from 754 ± 64.9 in the case of vehicle to 386 ± 51.4, which is a statistically significant reduction. MK300 and MK400 also reduced the SMA count to 540 ± 29 and 295 ± 34 respectively, as compared to vehicle control, which were statistically significant reductions [Figure 1].
|Figure 1: The total count of spontaneous motor activity in photoactometer. *P < 0.05, **P < 0.01, and ***P < 0.001. MK-Murraya koenigii; DW-Distilled Water|
Click here to view
Diazepam 5 mg/kg increased the open arm incursions from 29 ± 8.6 as compared to 53 ± 15, which is a statistically significant increase. However, MK300 and MK400 reduced the open arm count to 16 ± 7 and 10 ± 3.9 respectively, as compared to vehicle control, the latter being statistically significant [Figure 2]. Diazepam 5 mg/kg and vehicle produced similar closed arm counts of 191 ± 6.9 and 196 ± 19 respectively. MK200 to MK400, increased the closed arm count in a dose-dependent manner, the values were however not significant (223 ± 21.1, 236 ± 20 and 241 ± 6.4 respectively).
|Figure 2: The open arm count in elevated plus maze test. *P < 0.05; MK-Murraya koenigii; DW-Distilled Water|
Click here to view
Fluoxetine, 20 mg/kg reduced the duration of immobility in forced swim test from 145.5 ± 29 in the case of vehicle to 83 ± 51.4, which is a statistically significant reduction. MK400 also reduced the duration of immobility to 91 ± 17.3 as compared to vehicle control that were again statistically significant reductions [Table 1].
|Table 1: Effects of Murraya koenigii on experimental models of depression|
Click here to view
Fluoxetine, 20 mg/kg reduced the duration of immobility in the tail suspension test from 191.7 ± 9.6 in the case of vehicle to 76.5 ± 24, which is a statistically significant. MK also produced a dose-dependent decrease in the duration of immobility; however, it was statistically significant only in the case of MK400 (125 ± 17.4) [Table 1].
| Discussion|| |
The cost of modern anti-anxiety and anti-depressant drugs makes them unaffordable for the common man, especially in the Indian sub-continent where maximum population is poor. These drugs cannot be used for prolonged period because of their adverse effects. There is a delayed onset of action especially in patients of depression and high recurrence and relapse rates with these medicines on discontinuation.
A large number of patients either achieve partial or no response with these medicines even when used as multiple drug therapy. WHO recommends use of cheap and safe medicines of herbal origin. Hence, there is a need to develop more effective, cheaper and safer alternatives. This study may provide leads for newer agents by screening for activity in Murraya koenigii leaf extract.
Murraya koenigii is a tropical to sub-tropical plant belonging to family Rutaceae, which is native to India. Steam distillate of the curry leaves can be used as stomachic, purgative, febrifuge, and anti-emetic. Murraya koenigii also possesses antioxidant properties. Studies in both humans and animals have shown a strong correlation between anxiety and oxidative stress.,, Individuals who suffer with MDD display lower serum/plasma total antioxidant potentials, and reduced brain Glutathione (GSH) levels as compared to matched controls.,
In a study with an ethanol extract of Murraya koenigii leaves, it exhibited CNS stimulating and analgesic activities inin vivo models. In other studies with Murraya koenigii analgesic and anti-inflammatory activity has been reported.,
Not much research has been done to evaluate the role of Murraya koenigii in depression and anxiety, though antioxidant activity and other activities have been shown in previous studies.
Appropriate animal models of anxiety and depression were chosen for evaluation. These models are reliable and validated in large number of studies.
MK300 and MK400 reduced the SMA count as compared to vehicle control, which are statistically significant reductions. Diazepam, a long acting benzodiazepine significantly reduced the exploratory behaviour of mice in photoactometer, an indicator of sedative and anxiolytic effect. Murraya koenigii, also reduced this behavior in a dose-dependent manner pointing towards a sedative and/or anxiolytic effect.
Diazepam, as expected, increased the open arm incursion count, a measure of its anxiolytic effect. However, MK300 and MK400 reduced the open arm incursions, suggesting a sedative effect rather than an anti-anxiety effect. This sedative action is also suggested by the behaviour in SMA paradigm. A previous study, has paradoxically shown a central excitatory effect.
Diazepam 5 mg/kg and the vehicle group, produced similar closed arm count, whereas MK did produce a dose-dependent increase in the count which was, however, not statistically significant. This result suggests that MK is either producing a sedative effect or has no significant anti-anxiety like activity.
Fluoxetine, 20 mg/kg and MK400 reduced the duration of immobility in forced swim test which are statistically significant reductions as compared to vehicle control.
MK also produced a dose-dependent decrease in the duration of immobility in the tail suspension test; however, it was statistically significant only in the case of MK400.
Forced swim test and tail suspension test are validated primary screening methods producing despair behavior in mice. The agents that inhibit despair behavior in animals have shown antidepressant activity in humans. Fluoxetine, a selective serotonin reuptake inhibitor, reduced the duration of immobility (i.e., despair behavior) in both the models of our study. Murraya koenigii also reduced the duration of immobility (i.e., despair behavior) in both the models of our study, though only at the highest dose. This signifies that Murraya koenigii has an anti-depressant like activity. The effect is equivalent to fluoxetine in FST and less marked in the TST.
| Conclusion|| |
In two experimental animal models of despair behavior Murraya koenigii reduced the despair behavior, suggesting a putative anti-depressant like activity. In animal models for anxiety, the results were ambiguous. Murraya koenigii, reduced spontaneous locomotor activity in a dose-dependent manner pointing towards a sedative and/or anxiolytic effect. But in the elevated plus maze test, the result did not show any anxiolytic effect. Further studies are warranted to confirm the above results.
Financial support and sponsorship
Conflicts of interest
There are no conflicts of interest.
| References|| |
Bandelow B, Michaelis S. Epidemiology of anxiety disorders in the 21st
century. Dialogues Clin Neurosci 2015;17:327-35.
Bromet E, Andrade LH, Hwang I, Sampson NA, Alonso J, de Girolamo G, et al.
Cross-national epidemiology of DSM-IV major depressive episode. BMC Med 2011;9:90.
Ionescu DF, Rosenbaum JF, Alpert JE. Pharmacological approaches to the challenge of treatment-resistant depression. Dialogues Clin Neurosci 2015;17:111-26.
Ramkissoon JS, Mahomoodally MF, Ahmed N, Subratty AH. Antioxidant and anti-glycation activities correlates with phenolic composition of tropical medicinal herbs. Asian Pac J Trop Med 2013;6:561-9.
Mitra E, Ghosh AK, Ghosh D, Mukherjee D, Chattopadhyay A, Dutta S, et al.
Protective effect of aqueous curry leaf (Murraya koenigii
) extract against cadmium-induced oxidative stress in rat heart. Food Chem Toxicol 2012;50:1340-53.
Devatkal SK, Thorat PR, Manjunatha M, Anurag RK. Comparative antioxidant effect of aqueous extracts of curry leaves, fenugreek leaves and butylated hydroxytoluene in raw chicken patties. J Food Sci Technol 2012;49:781-5.
Krolow R, Arcego D, Noschang C, Weis S, Dalmaz C. Oxidative imbalance and anxiety disorders. Curr Neuropharmacol 2014;12:193-204.
Bouayed J, Rammal H, Younos C, Soulimani R. Positive correlation between peripheral blood granulocyte oxidative status and level of anxiety in mice. Eur J Pharmacol 2007;564:146-9.
Black CN, Bot M, Scheffer PG, Cuijpers P, Penninx BW. Is depression associated with increased oxidative stress? A systematic review and meta-analysis. Psychoneuroendocrinology 2015;51:164-75.
Porsolt RD, Bertin A, Jalfre M. Behavioral despair in mice: A primary screening test for antidepressants. Arch Int Pharmacodyn Ther 1977;229:327-36.
Steru L, Chermat R, Thierry B, Simon P. The tail suspension test: A new method for screening antidepressants in mice. Psychopharmacology (Berl) 1985;85:367-70.
Pellow S, Chopin P, File SE, Briley M. Validation of open: Closed arm entries in an elevated plus-maze as a measure of anxiety in the rat. J Neurosci Methods 1985;14:149-67.
Bonde SD, Nemade LS, Patel MR, Patel AA. Murraya koenigii
(Curry leaf): Ethnobotany, phytochemistry and pharmacology – A review. Int J Pharm PhytopharmacolRes 2011;1:23-7.
Zahin M, Aqil F, Husain FM, Ahmad I. Antioxidant capacity and antimutagenic potential of Murraya koenigii
. Biomed Res Int 2013;2013:263509.
Bondy SC. The relation of oxidative stress and hyperexcitation to neurological disease. Proc Soc Exp Biol Med 1995;208:337-45.
Radwańska-Wala B, Buszman E, Druzba D. Reactive oxygen species in pathogenesis of central nervous system diseases. Wiad Lek 2008;61:67-73.
Bouayed J, Rammal H, Soulimani R. Oxidative stress and anxiety: Relationship and cellular pathways. Oxid Med Cell Longev 2009;2:63-7.
Sarandol A, Sarandol E, Eker SS, Erdinc S, Vatansever E, Kirli S, et al.
Major depressive disorder is accompanied with oxidative stress: Short-term antidepressant treatment does not alter oxidative-antioxidative systems. Hum Psychopharmacol 2007;22:67-73.
Maes M, Mihaylova I, Kubera M, Uytterhoeven M, Vrydags N, Bosmans E, et al.
Increased 8-hydroxy-deoxyguanosine, a marker of oxidative damage to DNA, in major depression and myalgic encephalomyelitis/chronic fatigue syndrome. Neuro Endocrinol Lett 2009;30:715-22.
Brind L, Misra A, Srivastava S. Evaluation of central nervous system stimulating and analgesic activities of Murraya koenigii
leaves. J Acute Med 2014;4:81-5.
Darvekar VM, Patil VR, Choudhari AB. Anti-inflammatory activity of Murraya koenigii
Spreng on experimental animals. J Nat Product Plant Resour 2011;1:65-9.
Das BN, Biswas BK. Analgesic activity of the leaf extract of Murraya koenigii
. Int J Compr Pharm 2012;3:1-3.
[Figure 1], [Figure 2]