Evaluation of the effect of Wheatgrass powder on Stress-induced depression and memory loss in mice
Jangam Divya Latha1, Iswarya Obilineni2, A V S Ravi Sai Nadh2,
Vadivelan Ramachandran3, C. Sahana Reddy4, Kanuri.Bhuvaneswari5, Padavala Harika5, Tripuraneni Sai Praneeth5
1Assistant Professor, A.S.N Pharmacy College, Tenali, Guntur dt.
2Assistant Professors, KVSR Siddhartha College of Pharmaceutical Sciences, Vijayawada.
3Professor, Department of Pharmacology, JSS College of Pharmacy,
JSS Academy of Higher Education and Research, Ooty, Nilgris, Tamil Nadu, India.
4Research Scholar, JSS College of Pharmacy, JSS Academy of Higher Education and Research,
Ooty, Nilgris, Tamil Nadu, India.
5B. Pharmacy Students, KVSR Siddhartha College of Pharmaceutical Sciences, Vijayawada.
*Corresponding Author E-mail: divyalatha7777@gmail.com, aishwaryauday4@gmail.com
ABSTRACT:
Aim: To evaluate the effect of wheat grass powder on stress induced depression and memory loss in mice. Objective: The main objective is to observe the effect of wheat grass powder on stress induced depression and memory loss. Wheatgrass is the young grass of Triticum aestivum. The powder of wheat grass is known to contain chlorophyll, aminoatcids, minerals, flavonoids, vitamins etc. Stress was induced in mice by forced swim test. Methods: Forced swim test to induce stress, Behavioural tests by using actophotometer and Y maze apparatus and histopathological studies. Results: wheat grass powder treated group showed significant increase in locomotor activity when observed in actophotometer. Wheat grass powder treated group chosen and spent much time in novel arm of Y maze when compared to other groups. Conclusion: From result, it was concluded that wheat grass powder shows significant antidepressant activity and memory improving ability.
KEYWORDS: Depression, Memory loss, Stress induced, Forced swim test, Memory improvement.
INTRODUCTION:
Depression is a problematicheterogenousdisorder worldwide.1,2 Depression can be a gradual withdrawal from your active life and enjoyment of living.3 Patients suffering from this disorder feel empty, helpless and restless.4 It is caused by many factors like hereditary, stroke, parkinsonism, stress5 andunhealthy lifestyle like cigarette smoking, drug abuse6. Stressful events in life for long time results in many behavioural changes like biased cognition, major depressive disorder etc7. Major depressive disorder is a major psychological disorder affecting the quality of life8 of more than 17% of population worldwide9, hence it is described as psychic cold10.
According to WHO, in elders the overall prevalence rate of depressive disorder is 10-20%11 due to their varied medical conditions.12 Depression became a life threatening illness during this century,13 hence it is needed to be treated during the early symptoms14 and ultimately results in increased vulnerability to stress.15 Wheat grass is the young grass of the plant Triticum aestivum (wheat plant).16 Wheat grass is known to contain several phytochemical constituents like chlorophyll, saponins, alkaloids, phytosterols17, several enzymes like protease, amylase and lipase.18 Wheat grass also contains high amounts of calcium, phosphorus, potassium, magnesium, boron, zinc and molybdenum.19 Due to the presence of higher amounts of chlorophyll, it supplies more oxygen to all the tissues. Hence it is also known as Green Blood20 Chlorophyll present in the wheat grass powder helps in wound healing.21 Wheat grass powder is believed to be a effective detoxifier.22 It is well known of having anti-ulcer, anti-oxidant and anti-cancer activities.23 It is also known to increase the metabolic rate.24 Not only these, wheat grass powder helps the people by nourishing the kidneys and liver.25 Due to the many advantages of wheat grass powder, it was made available commercially in tablets form.26
MATERIALS AND METHODS:
Requirements:
Wheat grass powder (marketed product), Piracetam, Fluoxetine.
Instruments: UV-Visible Spectrophotometer-Thermo scientifics (Evolution-201), Cold centrifuge-Thermo scientifics (Sorvall ST8R), Shimadzu weighing balance-ATX/ ATY series, Photoactometer, Homogenizer- Remi (RQT-124 A/D), Advance Rotary Microtome- Medimeas Instrument –MRM 112
Animals: Male Swiss Albino mice weighing approximately 25-35g were used for anti-depressant activity and mice are used for the acute toxicological studies. The animals were housed at ambient temperature (24±2⁰C,) relative humidity (70%) and 12:12hrs light/dark cycle. Animal had free access to standard pellet diet and water ad libitum. The protocol approved by the Institutional Animal Ethics Committee (IAEC). Experimental design:
Animals are divided into 5 groups of ten mice each and the treatment was given once a day upto 1 week.
Group-1: Control
Group-2: Stress- induced group.
Group-3: Stress +Piracetam (200mg/kg) i.p27.
Group-4:Stress +Fluoxetine(10mg/kg) - oral +0.5%CMC.28
Group-5:Stress +Wheat grass powder(100mg/kg)
Procedure:
Forced swim test29
Mice are pre-tested by using y-maze and photoactometer and kept in a cylinder and forced to swim in water maintained at 25̊ C. Mice are placed in the cylinder for the first time are initially highly active, always trying to climb the wall or diving to the bottom. After 2-3min, the mice becomes slightly inactiveand after 5-6 min, the mice stops floating. After 15min, the mice were removed from the waterand allowed to dry in a heated enclosure (32̊ C) before being returned to their home cages. After 24hrs, they are placed in the cylinder again and the total duration of immobility is measured during a 5min test.Test drugs or standard are administered one hour prior to testing.30,31
Behavioral tests:
Mice of either sex were randomly divided into 5 groups of six each. The mice were placed individually inside the chamber of actophotometer for 10 min and 1stday basal activity score was noted for all group of animals. At stage of kindling, mice are placed again in actophotometer for 10 min and the activity was monitored. The average scores before and after drug administration are noted and the difference in the average activity scores were calculated for all the groups.
Y-maze test33
The Y-maze apparatus consists of three arms joined in the middle to form a ‘‘Y’’ shape. This test is based on the rodent’ curiosity to explore novel areas. Mice wereplaced in the start arm of the Y maze.The number of entries into and the time spent in each arm, and the first choice of entry were observed. During the training, rats were placed in the start arm in the first trial. Mice were allowed to explore the start arm and familiar arm which was opened, whereas the novel arm was closed. In the second trial, rats were allowed to explore the three arms; the number and order of arm entries for total 6 min duration was recorded. Total number of arm entries were used to calculate the spontaneous alteration behavior.
RESULTS:
Table 1: Effect of WGPon locomotor activity in actophotometer in stress induced mice in FST
|
Groups |
% increase in locomotor activity (Mean±SEM) |
|
Control |
6.377±1.585 |
|
Stress |
-23.98±5.952*** |
|
Piracetam |
9.434±1.962@@@ |
|
Fluoxetine |
14.01±1.049@@@ |
|
WGP |
11.07±1.509@@@ |
Values are expressed as Mean±SEM, n=10. One Way ANOVA test, *P<0.05, **P<0,01 and ***P<0,001 considered as statistically significant difference when compared to control and @P<0.05,@@<0,01 and @@@P<0,001compared to stress group.
Fig 1: Effect of WGP on locomotor activity in actophotometer in stress induced mice in FST.
Locomotor activity showed significant decrease in stress group when compared to control. Pretreated with Piracetam, Fluoxetine and Wheat grass powder group shown increase in locomotor activity .
Y-maze:
Locomotor activity of animals is expressed as (Mean±SEM), number of arm entries of ten mice challenged with the Y-Maze test within 5min of observation.
Table 2: Effect of WGP on Transfer latency time in Y maze test in stress induced mice in FST.
|
Groups |
Before treatment Mean+SEM |
After treatment mean+SEM |
|
Control |
3.200±0.374 |
3.000±0.4472 |
|
Stress |
3.000±0.4472 |
4.800±0.3742*** |
|
Piracetam |
3.000±0.3162 |
3.000±0.4472 |
|
Fluoxetine |
3.000±0.3162 |
3.200±0.3742 |
|
WGP |
3.200±0.3742 |
3.000±0.7071@@@ |
Values are expressed as Mean±SEM, n=10. One Way ANOVA test, *P<0.05, **P<0,01 and ***P<0,001 considered as statistically significant difference when compared to control and @P<0.05,@@<0,01 and @@@P<0,001compared to stress group
Fig 2:Effect of WGP on Transfer latency time in Y maze test in stress induced mice in FST.
Effect of WGP on number of arm entries in Y maze test:
Table 3: Effect of Wheat. G.P. on number of arm entries in Y maze test in stress induced mice in FST.
|
Groups |
Before treatment (Mean±SEM) |
After treatment (Mean±SEM) |
|
Control |
3.100±0.1528 |
3.300±0.3055 |
|
Stress |
3.033±0.088 |
1.600±0.5686 |
|
Piracetam |
3.400±0.1528 |
3.800±0.650 |
|
Fluoxetine |
3.100±0.3215 |
3.937±0.0917 |
|
WGP |
4.033±0.2186 |
4.900±0.100 |
One Way ANOVAtest, *P<0.05, **P<0,01 and ***P<0,001 considered as statistically significant difference when compared to control and @P<0.05,@@<0,01 and @@@P<0,001compared to stress group.
Stress group showed decreased locomotor activity than control group at 7th day of study. Whereas, after treatment with Piracetam, Fluoxetine and Wheat grass powder improved the locomotor activity.
The effect of WGP with duration of time in each arm in Y-maze test:
Locomotor activity of animals is expressed as mean duration of time in each arm (±SEM) of ten mice challenged with the Y-maze test within 10 min of observation.
Table 4: Effect of WGP on duration of time spent in novel arm in Y-maze:
|
Groups |
Before treatment (Mean+SEM) |
After treatment (Mean+SEM) |
|
CONTROL |
81.25±4.270 |
88.50±4.717 |
|
STRESS |
92.50±4.787 |
12.25±2.250*** |
|
PIRACETAM |
95.75±4.049 |
105.0±5.000@@@ |
|
FLUOXETINE |
94.50±2.630 |
92.50±4.787@@@ |
|
WGP |
110.0±13.54 |
120.00±0.00***, @@@ |
Values are expressed as Mean±SEM, n=10. One Way ANOVA test, *P<0.05, **P<0,01 and ***P<0,001 considered as statistically significant difference when compared to control and @P<0.05,@@<0,01 and @@@P<0,001compared to stress group.
Fig 4: Effect of WGP on duration of time in Y maze test in stress induced mice in FST.
In Control group duration of time increases in novel arm, whereas in stress group mice showed decreased locomotor activity. Pre-treated groups showed increased duration of time in novel arm. WGP treated group has showed better results against compared to all groups
Effect of Wheat.G.P. on first choice of novel arm:
Table 5: Effect of WGPon % of first choice of novel arm in Y maze test in stress induced mice in FST.
|
S. No |
Groups of animal |
Mean±SEM |
|
1 |
CONTROL |
83.33±16.67 |
|
2 |
STRESS |
16.67±16.67** |
|
3 |
PIRACETAM |
100.0±0.00@@ |
|
4 |
FLUOXETINE |
83.33±16.67@@ |
|
5 |
WGP |
100±0.0@@ |
Values are expressed as Mean±SEM, n=10. One Way ANOVA test, *P<0.05, **P<0,01 and ***P<0,001 considered as statistically significant difference when compared to control and @P<0.05,@@<0,01 and @@@P<0,001compared to stress group.
Fig 5: Effect of WGPon % offirst choice of novel arm in Y maze test in stress induced mice in FST.
Before treatment control group of all animals were chosen novel arm but stress group does not choose the novel arm entry. Pre-treatment with Piracetam, Fluoxetine and wheat grass powder chosen the novel arm when compared to all groups except control group.
DISCUSSION:
In Forced swim test, the mice pre-treated with fluoxetine, piracetam and wheat grass powder showed improved climbing time and swimming time. Decreased in immobility time was observed in sub chronic treatment with fluoxetine. Fluoxetine, antidepressant decreased the immobility time (Floating time) in FST test34,35. Scientific reports on Piracetam in FST test evident that, it also decreased the immobility time in FST test36,37 WGP shown Anti-immobility activity in FST test and the results were comparable to that of standard fluoxetine group.
Piracetam improved FST induced stress produced changes in Y-maze test38,39like decreased transference latency, % first choice of novel arm, duration of time spent in novel arm. Fluoxetine showed memory improvement in FST test. WGP treatment showed significant differences in novel arm entries, duration of time spent in the novel arm, transfer latency compared to stress group.
CONCLUSION:
WGP 100 mg/kg pre-treatment shown protective effect against FST induced depression and memory loss.
REFERENCES:
1. Jaseela KP. Anjan K. Veeresh B. D. NarayanaSVB. Studies on Anti-depressant and Anti-inflammatory Activities of Curcuma aromatica Rhizome in Experimental Animal Models. Asian J. Pharm. Res. 2016; 6(2): 79-86.DOI:10.5958/2231-5691.2016.00014.9
2. Sumanpreet K. A Cross-sectional Study to Assess the Psychosocial Deviations and Depression among the Senior Citizens of Selected Rural Community of Gurdaspur, Punjab. Asian J. Nur. Edu. and Research. 2016; 6(3): 401-406.DOI: 10.5958/2349-2996.2016.00075.6
3. Mulsant BH. Ganguli M. Epidemiology and diagnosis of depression in late life. The Journal of Clinical Psychiatry. 1999 Aug 1;60(suppl 20):9-15. PMID: 10513852
4. Juna T. VeereshBD. Anjan K.Narayana SB. Studies on Anti-Depressant Activity of Perment-A Polyherbal Formulation in Experimental Animal Models. Research J. Pharmacology and Pharmacodynamics. 2016; 8(2): 45-54.DOI: 10.5958/2321-5836.2016.00009.4
5. Praveen KU. SwarnaLatha M. Shashidhar RR.Chakravarthi G. Evaluation of anti-depressant activity of Methanolic Seed Extract of Avenasativa L. In Mice. Research J. Pharmacology and Pharmacodynamics. 2013; 5(4). 212-217.DOI:10.5958 2321-5836
6. Sumanpreet K. A Descriptive Study to Assess Depression and Codependency among Wives of Alcoholics in a Selected Rural Community of Gurdaspur. Punjab. Asian J. Nur. Edu. and Research. 2016; 6(2): 183-187. DOI: 10.5958/2349-2996.2016.00033.1
7. Gal RL. Lin X. How could stress leads to Major depressive disorder. IBRO Reports. 2018; 4: 38-43. DOI: 10.1016/j.ibror.2018.04.001
8. Longfei Y. Yinghao Z. Yieun W. Lei L. Xingyi Z. Bingjin L Ranjicui. The effects of psychological stress on depression. Current Neuropharmacology. 2015; 13(4): 494-504. DOI: 10.2174/1570159X1304150831150507
9. Fushun W. Jiongjiong Y. Fang P. James AB and Jason H. Early life stress and Depression, Frontiers in psychiatry Mood and Anxiety Disorders 2020. Doi.org/10.3389/fpsyt 2019.00964.
10. Fateme GH. Saeid P. Mortaza. RoohollahI. The relationship between religiosity and depression among Medical students. Asian J. Nur. Edu. and Research. 2016; 6(3): 414-418. DOI: 10.5958/2349-2996.2016.00078.1
11. Sangroyangla, T. TemjenyanglaI. Tsungrosenla L. Vizayieno. KhumjanbeniM. Prevalence of Depression and the associated risk factors among the Elderly. Asian J. Nursing Education and Research. 2019; 9(4): 552-554. DOI: 10.5958/2349-2996.2019.00119.8
12. Anisha V.Femila.P. Santhi A. A Study to Assess the Effectiveness of Taichi Exercise on Depression among Senior Citizens in Selected Old Age Home. Asian J. Nur. Edu. and Research. 2014; 4(4): 403-407. DOI: 10.5958/2349-2996
13. MalarkodiV.Ravichandiran.V.Ramamoorthy.S.Vijayalakshmi.A.Srikanth.J. Antidepressant-Like Effects of the Ethanolic Extract of Albizzialebbeck (Linn) Leaves in Animal Models of Depression. Research J. Pharmacognosy and Phytochemistry. 2010; 2(1): 30-33.DOI: 10.1007/7854_2010_108
14. Kavitha.V.Leethyal.R. Prevalence of postnatal depression among postnatal mothers. Asian J. Nur. Edu. and Research. 2012; 2(1): 33-36. DOI: https://doi.org/10.22159/ajpcr.2016.v9i6.12900
15. Gustavo ET. Charles BN. The link between stress and depression: Psychoneuroendocrinological, Genetic and Environmental interactions,The journal of Neuropsychiatry and Clinical Neurosciences. 2015. DOI: 10.1176/appi.neuropsych.15030053
16. Rajesh M. Ramesh B. A study on wheat grass and its nutritional value. Food Science and Quality Management. .
17. Devi SK. Hari PK. Nalini GR. Veenaneesh P. Ravichandra S. Wheat grass juice, Triticum aestivum Linn, A therapeutic tool in pharmaceutical research,an overview. IJPPR. 2015; 3:112-121.
18. Mujoriya R. Bodla RB. A study on wheat grass and its nutritional value,Food science and Quality Management. 2011; 2:1-9.
19. Swati P. Sushma D. Indira R. Alka G. Mamta D. Multitude potential of wheat grass juice (Green Blood): An overview. Chronicles of Young Scientist. 2010: 1(2): 23-28.
20. RuchaDG, SangeetaJP, AnirudhaRB, Wheat grass: Green blood can help to fight cancer.Journal of Clinical and Diagnostic Research, 2017; 11(6): ZC40-ZC42 doi: 10.7860/JCDR/2017/26316.10057
21. Bowers W. Chlorophyll in wound healing and supportive disease. Am. J. Surg. 1947:73:37-50.
22. MukthiT. Seshamma S. SailajaN.A mini review on wheat grass. Research and Reviews; Journal of Pharmacognosy and Phytochemistry.
23. N.Singh. P.Verma. B.R.Pandey. Therapeutic Potential Of Organic Triticum Aestivum Linn. (Wheat Grass) In Prevention And Treatment Of Chronic Diseases; An Overview. International Journal of Pharmaceutical Sciences and Drug Research. 2012; 4(1).
24. Rimple MKK. Kumar R. Newton A. Reeta SL. Polypharmacological effects of green blood therapy: An update.World Journal of Pharmaceutical and Medical Research. 2016; 2(1): 10-21.
25. S.Dey. R.Sarkar. P.Ghosh.R.Khatun. K.Ghoral. R.Choudhury. Effect of wheat grass juice in supportive care of terminally ill cancer patients- A teritiary Cancer centre experience from India. Journal of Clinical Oncology. 2006: 8634-8634.
26. S.D. Kulkarni. R. Acharya. N.S. Rajurkar. A.V.R. Reddy. Bioaccessibility of some elements from wheat grass (Triticumaestivum L) by invitro gastrointestinal digestion combined with neutron activation analysis with ammonium bicarbonate as an alternate base. Food Chemistry. 2006; 95(4): 699-707.
27. Moran TH. Capone GT. Knipp S. Davisson MT. Reeves RH. Gearhart JD. The effects of piracetam on cognitive performance in a mouse model of Down's syndrome. Physiology and Behavior. 2002 Nov 30; 77(2): 403-9 DOI: 10.1016/s0031-9384(02)00873-9
28. Sirisha G. Rahul PB. Usha MD. Evaluation of antidepressant effect of chronic administration of tramadol alone and in combination with fluoxetine in low doses in albino mice. Int J Pharm Pharm Sci. 2014; 6(6): 101-5DOI: 10.13040/IJPSR.0975-8232.6(6).2462-67.
29. Vogel H. Drug discovery and evaluation: pharmacological assays. Springer Science and Business Media. 2007 Oct 30.
30. Mezadri TJ. Batista GM. Portes AC. Marino-Neto J. Lino-de-Oliveira C. Repeated rat-forced swim test: reducing the number of animals to evaluate gradual effects of antidepressants. Journal of Neuroscience Methods. 2011 Feb 15; 195(2): 200-5 DOI: 10.1016/j.jneumeth.2010.12.015.
31. Petit-DB. Chenu F. Bourin M. Forced swimming test in mice: A review of antidepressant activity. Psychopharmacology. 2005 Jan 1; 177(3): 245-55 DOI: 10.1007/s00213-004-2048-7
32. Verma A. Jana GK. Sen S. Chakraborty R. Sachan S. Mishra A. Pharmacological evaluation of Saracaindica leaves for central nervous system depressant activity in mice. J Pharm Sci Res. 2010; 2(6): 338-43.
33. Detrait E. Brohez C. Hanon E. Ryck M. Automation of continuous spontaneous alternation to increase the throughput for in vivo screening of cognitive enhancers, Optimization of the Ethovision system for the Y-maze test in mice. Proceedings of Measuring Behavior. 2010 Aug 24: 141-144.
34. Bianchi M. Moser C. Lazzarini C. Vecchiato E. Crespi F. Forced swimming test and fluoxetine treatment: in vivo evidence that peripheral 5-HT in rat platelet-rich plasma mirrors cerebral extracellular 5-HT levels, whilst 5-HT in isolated platelets mirrors neuronal 5-HT changes. Experimental Brain Research. 2010 Mar 1; 143(2): 191-7 DOI: 10.1007/s00221-001-0979-3.
35. Reneric JP. Lucki I. Antidepressant behavioral effects by dual inhibition of monoamine reuptake in the rat forced swimming test. Psychopharmacology. 2009 Mar 1; 136(2): 190-7 DOI: 10.1007/s002130050555.
36. Gokhale VS. Bhide SS. Jalgaonkar SV. Marathe PA. Mane Y. Khan FM. Rege NN. Evaluation Of Effect Of Piracetam In Experimental Models Of Depression.International Journal of Pharmaceutical Sciences and Research. 2013 Jul 1; 4(7): 2667 DOI: 10.13040/IJPSR.0975-8232.4(7).2667-72.
37. Kale PP, Addepalli V, Sarkar A, Patel S, Savai J. The combination of antidepressant duloxetine with piracetam in mice does not produce enhancement of nootropic activity. Experimental Neurobiology. 2014 Sep 1; 23(3): 224-30 DOI: 10.5607/en.2014.23.3.224.
38. Uma G, Maheswari S, Neuroprotective effects of polyherbal formulation (Indian Noni) on scopolamine-induced memory impairment in mice. International Journal of Pharmaceutics. 2014; 6: 354-7.
39. Poornachandra MN, Khanam S, Shivananda BG, Shivananda TN, Dris R. Mucunapruriens (L.) DC-A novel drug for learning and memory retrieval. Journal of Food Agriculture and Environment. 2015 jul; 3(3/4): 13.
Received on 26.10.2020 Modified on 08.06.2022
Accepted on 18.01.2023 © RJPT All right reserved
Research J. Pharm. and Tech 2024; 17(5):2315-2319.
DOI: 10.52711/0974-360X.2024.00363