|Year : 2014 | Volume
| Issue : 4 | Page : 231-236
Adaptogenic and nootropic activity of Mimosa pudica in albino wistar rats
Sibi Perumbamkudiyil Ittiyavirah1, Ibrahim Pullochal2
1 Department of Pharmacology, Neuropharmacology Research Division, Mahatma Gandhi University, Kottayam, Kerala, India
2 Research Scholar, University College of Pharmacy, Cheruvandoor Campus, Mahatma Gandhi University, Kottayam, Kerala, India
|Date of Submission||17-Mar-2014|
|Date of Acceptance||18-May-2014|
|Date of Web Publication||22-Aug-2014|
Sibi Perumbamkudiyil Ittiyavirah
Department of Pharmacology, Neuropharmacology Research Division, University College of Pharmacy, Cheruvandoor Campus, Mahatma Gandhi University, Kottayam 686 631, Kerala
Source of Support: None, Conflict of Interest: None
| Abstract|| |
Aim: To evaluate the adoptogenic activity of ethanolic extract of Mimosa pudica L. in chronic Alzheimer's model. Materials and Methods: The dried whole part of the plant were subjected to hot continuous extraction method using ethanol as solvent and were standardized using pharmacognostical and phytochemical screening. The in-vitro antioxidant studies were conducted by super oxide scavenging assay and hydrogen peroxide scavenging assay. Satisfactory IC 50 values were obtained for both assays. Dose selection for in-vivo study was made on the basis of already conducted acute toxicity studies (500 mg/kg body weight). Oral administration of extract of M. pudica was continued for 21 consecutive days. Adaptogenic activity was assessed by using oral dose of 500 mg/kg of EEMP and 2 mg/kg diazepam as test and standard compound respectively. Results: There was a significant improvement in memory, which was observed from the test paradigms viz., morris water maze, radial arm maze. Forced swim test was used as the observation paradigm for the adaptogenic activity and EEMP caused significant reduction in swimming endurance time. Conclusion: It was concluded that ethanolic extract of M. pudica at the dose of 500 mg/kg p.o produces potential changes in chronic Alzheimer's model and stress.
Keywords: Adaptogenic, Mimosa pudica, nootropic activity
|How to cite this article:|
Ittiyavirah SP, Pullochal I. Adaptogenic and nootropic activity of Mimosa pudica in albino wistar rats. Int J Nutr Pharmacol Neurol Dis 2014;4:231-6
|How to cite this URL:|
Ittiyavirah SP, Pullochal I. Adaptogenic and nootropic activity of Mimosa pudica in albino wistar rats. Int J Nutr Pharmacol Neurol Dis [serial online] 2014 [cited 2020 Aug 6];4:231-6. Available from: http://www.ijnpnd.com/text.asp?2014/4/4/231/139404
| Introduction|| |
Stress can be defined as the sum total of all the reactions of the body, which disturb the normal physiological condition and results in a state of threatened homeostasis. Normally stress-induced changes are compensatory, self-limiting and adaptive. However, in higher animals when stress events of any nature (Physical, Chemical, Biological, and Emotional) over certain 'threshold' limits, the changes become rather irreversible. It leads to altered homeostasis and exhaustion, manifesting itself in the pathologic form of stress-induced disease and maladjustment. There is no treatment in modern drug therapy for stress-related diseases. The available techniques for increasing endurance performance include physical training for endurance work, yogic and meditation practices, supplementation of neutraceuticals and intervention by adaptogens. Present study will provide a scientific base for experimental research on Indian herb for stress-related diseases. The term 'Adaptogen' denotes an agent that improves adaptation capacity of the organism during stress and "Antistress" agent is a pharmacological word for the same, meaning an agent, which nullifies or prevents ill effects of stress and improves adaptation. 
Alzheimer's disease (AD) is a slowly progressive disease of the brain that is characterized by impairment of memory and eventually by disturbances in reasoning, planning, language, and perception. Neurological disorders results in various degrees of disability and loss of productive life. 625 per 100000 population per year consult their family doctor regarding a neurological disorders, rates between males and females being similar.  Poor learning abilities and impaired memory are increasing their importance in accompanying with elderly population and stress exposure. Hence there is an urgent need for cognitive enhancer to develop. Recent studies showed that the oxidative stress play a major role in neurodegenerative disorders, and therefore the present study focuses on M. pudica due to its potent antioxidant activity. The reports of literature were further supported with its neuroprotective activity and sedative hypnotic activity. ,
Several naturally occurring herbs viz., Abana, Centella asiatica and Celastrus paniculatus were found to be have potent nootropic activities. ,, Piracetam and fulvestrant were evaluated for its memory enhancing activity. , Other plants having adaptogenic activity include Micania micrantha, Panex ginseng, Withania somnifera.
| Materials and methods|| |
Ethanol (Travancore chemicals), Piracetam, Diazepam (Nice chemicals (P), Ltd, Kochi, Kerala), D-galactose, Carboxy methyl cellulose (CMC), Hydrogen peroxide (Nice chemicals (P), Ltd, Kochi, Kerala), Gallic acid (Nice chemicals (P), Ltd, Kochi, Kerala), Methanol, Distilled water, Riboflavin, EDTA, Sodium hydroxide, potassium dihydrogen phosphate, Nitro blue tetrazolium (NBT), Soxhlet extractor (Glastron Laboratory Equipments, Haryana, India), UV/VIS spectrophotometer (Systronic double beam-UV-2201, systronics, India).
The whole plant of M. pudica was collected from nearby places of University College of Pharmacy Cheruvandoor and approved by Professor Jomy Augestine, head of Department of Botany, St. Thomas College, Palai. The plant was collected during the month January/February and dried under light shade at room temperature and subjected to extraction procedure.
The dried whole plant was grounded to get a free flowing powder. With the help of a soxhlet apparatus the dried powder were subjected to extraction using dehydrated alcohol . The extract obtained was filtered through Whatmann filter paper and dried at 40-50°C to get a blackish green semisolid mass, which was dissolved in saline solution for final use.
Albino wistar rats weighing 150-200 g of either sex maintained under standard husbandry conditions were used for the screening which was obtained from the animal house of the UCP. Animals were fed with standard lab food during the study period. The experiments were performed after getting the approval for the experimental protocol from the institutional animal ethical committee (IAEC) with the IAEC number (IAEC/004/MPH/UCP/CVR/13).
In vitro antioxidant activity
Hydrogen peroxide scavenging assay
One milliliter of the Ethanolic extract of Mimosa pudica (EEMP) and 0.6 ml hydrogen peroxide was added to the test tube and it was made upto 5 ml with phosphate buffer. The absorbance was measured at 230 nm using UV spectrophotometer against a blank solution containing phosphate buffer without using hydrogen peroxide. Assay was done in triplicate for all test samples and averages were counted. Percentage inhibition was determined by the given formula:
%inhibition = [(A 0 -A 1 )/A 0 ] ×100
were, A 0 was the absorbance of the control and A 1 was the absorbance of sample. 
Super oxide radical scavenging assay
0.02 ml of various concentrations of extract (12.5-200 μg/ml), 0.05 ml of Riboflavin solution (0.12 mM), 0.2 ml of EDTA solution [0.1 M], and 0.1 ml NBT (Nitro-blue tetrazolium) solution (1.5 mM) were mixed in test tube and reaction mixture was diluted up to 2.64 ml with phosphate buffer (0.067 M). The absorbance of solution was measured at 560 nm Dimethyl sulfoxide (DMSO) as blank after illumination for 5 min and difference in OD was determined after 30 min incubation in fluorescent light using UV visible spectrometer.
% Inhibition = [OD of control - OD of test sample/OD of control] ×100 
Evaluation of antistress activity
Swimming endurance test
In Forced swim test (FST) the animals were forced to swim in a restricted space from which they were unable to climb which has cause a state of depression and anxiety resulted in severe mental stress. During the test the animals were induced to characteristic behavior of immobility which reflected a state of despair resembling a mental depression and had been used to screen anti-depressants. The stress was created once daily for a period of 8 days and the animals were allowed to swim till complete exhaustion and the time was noted when the animals start drowning, which was noted as swimming endurance time. Finally the mean swimming endurance time was calculated. 
Group 1: Normal control (CMC 0.5%w/v, for a period of 42 days)
Group 2: Positive control (D-Galactose 100 mg/kg, for a period of 42 days)
Group 3: Standard group (D-Galactose 100 mg/kg, for a period of 21 days + Piracetam 100 mg/kg, for a period of next consecutive 21 days).
Group 4: Test group (D-Galactose 100 mg/kg, for a period of 21 days + EEMP 500 mg/kg, for a period of next consecutive 21 days.
Radial arm maze
The radial arm maze used was open type. It consists of a central circular arena and eight equally sized arms. Each arm consists of 60-cm length and 20-cm breadth. The animals were placed on the central platform and allowed to move freely on each arm. At far end of each arm a small plastic dish was kept and sucrose food pellet was mounted on it. The spatial working and spatial reference memory was evaluated by the instrument.
The animals are habituated to the environment, placed to the central platform and allowed to explore maze for 15 minutes. Reinforces (or baits) are scattered on the arms. For the training periods of first days all the arms were baited and followed by baiting of alternative arms. Retention latencies (time for rat to reach the reward) was recorded on 21 st and 42 nd day.
The spatial memory error was measured in the radial arm maze apparatus. The two types of spatial memory include spatial working memory and spatial reference memory. Spatial working memory error is considered as the double entries into the baited arm while the spatial reference memory error is considered as entries in never baited arm. 
Morris water maze
Morris water maze consisted of large circular pool (75-cm diameter and 30-cm height) filled with water at a depth of 20 cm. The pool was divided into four equal quadrants with the help of a thread. A circular platform was placed in one quadrant of the pool 1 cm above the water level during the acquisition phase. A similar platform was placed 1 cm below the water level during the retention phase. The position of the platform was not changed in any quadrant during assessing of both phases. Each animal was subjected to four consecutive trials with a gap of 5 min. Each animal was allowed 120 sec to locate the platform. If animal failed to reach the platform within 120 sec the animal was guided to reach the platform.
The control animals were treated with the D-galactose throughout the period of 42 days. The test drug was given to the animal on 20 th day onwards of treatment period. After 24 h the animals were released on the Morris water maze from the same quadrant of the maze. The animals were allowed to find the hidden platform and the time taken for the finding was recorded and termed as the first retention latency. This was repeated on 42 nd day which was the last day of drug treatment and the time taken for finding the hidden platform was recorded and which was termed as the second retention latency. 
Statistical analysis was done by using GRAPHPAD PRISM 5.0. All the values of Biochemical parameters and body weight were expressed as Mean ± Standard Error Mean (SEM). The values were analyzed for statistical significance using one-way analysis of variance (ANOVA).
| Result and Discussion|| |
The observations of preliminary phytochemical analysis on M. pudica showed the presence of alkaloids, flavonoids, steroids, tannins, and phenolics. In the present study the results of swimming endurance test showed that the M. pudica plant extract was found to be effective for its anti-stress activity as exhibited by the significant reduction in swimming endurance time [Figure 1]. A group of plant-based drugs, the adaptogen increase the non-specific resistance of the organisms against different types of stressors (Kokate 1994). Various measures are available to counteract the adverse effect of stress, which include pharmacological and non-pharmacological methods. Use of several anti-stress agents particularly, benzodiazepines like diazepam showed significant anti-stress activity against various models of stress. But the problems of tolerance and physical dependence exhibited by benzodiazepines on prolonged use, limits their utility.
|Figure 1: Evaluation of swimming endurance time of EEMP in forced swimming test|
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Debnath et al had reported that the plasma levels of adrenaline and noradrenaline are enhanced during stress induced by swimming endurance test. In addition, monoamine oxidase (MAO) levels in the brain were reportedly decreased during stress. It is postulated that the EEMP has the property to elevate the physical endurance and thereby causing the antistress effect. These results are in accordance with the findings that the normalizing the plasma levels of MAO and noradrenaline.
Stress is believed to act by increasing non-specific resistance. In-vitro antioxidant studies were carried out to assess the scavenging potential of M. pudica. Hydrogen peroxide scavenging assay and super oxide scavenging assay has been used to evaluate the free radical scavenging activity of natural antioxidant. The results obtained revealed the antioxidant activity and were shown in [Figure 2] and [Figure 3] respectively. In the present study the stressful environment in forced swim test leads to endogenous depression in animals and pretreatment with ethanolic extract of M. pudica had increased the stress tolerance indicating their adaptogenic/anti-stress activity and it may be due to elevation of succinate dehydrogenase (SDH) in the brain. This enzyme is responsible for utilization and conservation of energy in the cellular system of the organism, which helps adaptive processes during stress. In the present study the swimming endurance time was found to be more when compared to the control group.
|Figure 2: Comparison of IC50 values of ascorbic acid and EEMP in hydrogen peroxide radical scavenging|
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|Figure 3: Comparison of IC50 values of gallic acid and EEMP in superoxide radical scavenging|
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The present study also revealed its nootropic potential in various paradigms. In Morris water maze retention latencies were reduced siginifcantly (P < 0.001), where retention latency is the time taken by the rat to reach the hidden platform. The results were shown in [Figure 4] and [Figure 5]. In radial arm maze the number of errors were reduced significantly (P < 0.01), EEMP treated rats showed little errors in entering the arms and a less time to complete the whole session in the maze. The two parameters measured were working memory error (WME) and reference memory error (RME) on 21 st and 42 nd day of the study. The results for WME and RME on 21 st day were shown in [Figure 6] and [Figure 7], whereas for 42 nd day in [Figure 8] and [Figure 9] respectively. Time taken by the animal to complete one session in radial arm maze on 21 st day and 42 nd day were shown in [Figure 10] and [Figure 11]. These effects are may be suggested the effect of EEMP for the hippocampal repair or stimulate the existing neurons or it may also suggested that its effect to protect vulnerable neurons.
|Figure 10: Time taken to complete one session in radial arm maze- 21st day|
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|Figure 11: Time taken to complete one session in radial arm maze- 42nd day|
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| Conclusions|| |
The result from the study indicated that ethanolic extract of Mimosa pudica possessed significant anti-stress activity along with a potential protective effect against chronic Alzheimers model.
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[Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8], [Figure 9], [Figure 10], [Figure 11]