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 Table of Contents  
ORIGINAL ARTICLE
Year : 2014  |  Volume : 2  |  Issue : 1  |  Page : 29-35

Nootropic potential of Bauhinia variegata: A systematic study on murine model


1 Drug Discovery Laboratory, Ram Institute of Technology-Pharmacy, Jabalpur, Madhya Pradesh, India
2 Drug Discovery Laboratory, Ram Institute of Technology-Pharmacy, Jabalpur; Pranav Institute of Pharmaceutical Science and Research, Sitholi, Gwalior, Madhya Pradesh, India
3 Pharmaceutics Research Laboratory, Shri Ram Institute of Technology-Pharmacy, Jabalpur, Madhya Pradesh, India

Date of Web Publication4-Jun-2014

Correspondence Address:
Aditya Ganeshpurkar
Drug Discovery Laboratory, Shri Ram Institute of Technology - Pharmacy, Jabalpur, Madhya Pradesh - 482 002
India
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/2321-4848.133792

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  Abstract 

Objectives: Bauhinia variegata Linn (leguminosae) is one of the important medicinal herbs used traditionally to treat fever, as tonic, astringent, diarrhea, dysentery, hemorrhoids, piles, edema. Recent findings on Bauhinia variegata Linn have demonstrated its antioxidant, anti-hyperlipidemic, and hepatoprotective potential. The present work is focused to evaluate nootropic potential of Bauhinia variegata Linn in rats. Materials and Methods: The leaves of Bauhinia variegata were collected in the month of January from Jawaharlal Nehru Krishi Vishwavidyalaya Jabalpur (Madhya Pradesh). Leaves were subjected for isolation of crude flavonoids and characterized by total flavonoid content assay. Flavonoid-rich extract of Bauhinia variegata was studied for acute oral toxicity as per revised Organization for Economic Cooperation & Development guidelines No. 423. Nootropic activity was determined by elevated plus maze, rotating rod apparatus, baclofen-induced catatonia, diazepam-induced amnesia. Results: Flavonoid-rich fraction of Bauhinia variegata caused no alteration in locomotion in animals. In the current study, animals treated with flavonoid-rich fraction of Bauhinia variegata (400 mg/kg) showed a significant decrease in transfer latency as compared to the control group, which indicates cognitive enhancement effect flavonoid-rich fraction of Bauhinia variegata. In rota rod studies, flavonoid-rich fraction of Bauhinia variegata increased fall of time as compared to diazepam. In baclofen-induced catatonia, administration of flavonoid-rich fraction of Bauhinia variegata demonstrated protective effect on rats. Over all, flavonoid-rich fraction of Bauhinia variegata was found to enhance the performance of murine models. Conclusion: Thus, it could be concluded that flavonoids from Bauhinia variegata possess nootropic potential. However, more systematic studies are required to determine its exact mechanism of action.

Keywords: Amnesia, Bauhinia variegata, catatonia, memory, nootropic


How to cite this article:
Jatav N, Ganeshpurkar A, Gupta N, Ayachi C, Ramhariya R, Bansal D, Dubey N. Nootropic potential of Bauhinia variegata: A systematic study on murine model. Arch Med Health Sci 2014;2:29-35

How to cite this URL:
Jatav N, Ganeshpurkar A, Gupta N, Ayachi C, Ramhariya R, Bansal D, Dubey N. Nootropic potential of Bauhinia variegata: A systematic study on murine model. Arch Med Health Sci [serial online] 2014 [cited 2017 Apr 23];2:29-35. Available from: http://www.amhsjournal.org/text.asp?2014/2/1/29/133792


  Introduction Top


Recent years have seen a sudden surge in an array of mental illnesses such as dementia and other memory-related disorders. Memories are ability of an individual to record event, information and retain them over short or long period of time, recall the same whenever needed. Age and stress along with emotional breakdown are some conditions, which could lead to loss of memory and memory deficit disorder, with change in cognitive and behavioral attitude that may lead to progression of schizophrenia and Alzheimer's diseases. Dementia is one of the age-related mental problems, characterized by loss of intellectual ability and interferes with one's occupational and social activities. It is a characteristic symptom of Alzheimer's disease. [1]

Alzheimer's disease is a neurodegenerative brain disorder with slow onset that interferes with the patient's professional life, social activities, and relationships. [2] As discussed above, it causes dementia, personality changes, and ultimately death. [3] It primarily affects the geriatric population, and is accounted for 50-60% of dementia cases in persons over 65 years of age. [4]

Nootropics agents are known to augment the cognitive dexterity. [5],[6] Various drugs like diazepam, alcohol, barbiturates are known to have detrimental effect on learning and memory. Nootropic agents such as piracetam, [7] pramiracetam, aniracetam, [8] and cholinesterase inhibitors like onepezil are used primarily for improving memory, mood, and behavior. However, the resulting side-effects associated with these agents have limited their use. [9] Thus, it is necessary to explore the utility of traditional medicines in the treatment of various cognitive disorders.

Ayurveda is an ancient form of Indian medicine, which deals with use of plants and plant products for maintenance of good health. This indigenous form of medicinal system uses the active ingredients present in plants for treating various diseases. [10] Bauhinia variegata Linn (leguminosae) is known as Kanchanar in Hindi. It is a medium-sized tree abundant in Sub-Himalayan tract extending eastwards to Assam, Eastern, Central and South India. [11] Various parts of the plants viz., leaves, flower buds, flower, stem, stem bark, seeds, and roots are known for therapeutic activity and are used as tonic, astringent, for fever, diarrhea, dysentery, hemorrhoids, piles, edema, laxative, anthelmintic, antileprotic, in skin diseases, for wound healing, antigoitrogenic, antitumor, in obesity, stomatitis, antidote for snake poisoning, in dyspepsia, flatulence, and as carminative. [12]

Recent findings on Bauhinia variegata Linn have demonstrated its antioxidant, anti-hyperlipidemic, [13] and hepatoprotective potential. [14] The plant is also known to inhibit enzyme acetylcholinesterase. [15] Extract of Bauhinia variegata stem bark has shown protective action against milk-induced eosinophilia in mice. [16]

The present work is focused to evaluate nootropic potential of Bauhinia variegata Linn in rats.


  Materials and Methods Top


Plant material

The leaves of Bauhinia variegata were collected in the month of January from Jawaharlal Nehru Krishi Vishwavidyalaya Jabalpur (M.P.). The plant was identified and authenticated from Department of Crop and Herbal Physiology, by Dr. A. B. Tiwari, Sr. Scientist, Jawaharlal Nehru Krishi Vishwavidyalaya, Jabalpur (M.P.)

Drugs and chemicals

Bulk solvents like Petroleum ether (60 - 80 0 C), Benzene, Chloroform, Acetone, Ethyl alcohol (95%) Methanol, and routine chemicals were obtained from Central Drug House, India. All chemicals used were of analytical grade.

Preparation of Flavonoid-rich fraction

Briefly, One hundred g of air-dried leaves were grounded to fine powder and soaked in 70% ethanol for 24 h with continuous stirring. The soaked mixture was filtered using Whatmann No. 1 filter paper and the pellet discarded after centrifugation of the filtrate at 10,000 rpm at room temperature (25°C). The supernatant was concentrated in vacuo by means of a rota-vapor and then dissolved in as little water as possible and washed 3 times with chloroform. The resultant residual layer after extraction 3 times with ethyl acetate and subjected to concentration in vacuo served as the flavonoid-rich fraction (FRF). [17]

Estimation of flavonoids

Assay for total flavonoids content

Total flavonoid content was determined using the method given elsewhere. [18],[19] Briefly, Aluminum trichloride (1 ml, 2% w/v) in methanol was mixed with the same volume of the flavonoid-rich fraction of Bauhinia variegata (1 ml, 2000 μg/ml). Absorption readings at 415 nm were taken after 10 min against a blank sample consisting of a flavonoid-rich fraction of Bauhinia variegata (1 ml, 2000 μg/ml) with methanol (1 ml) without AlCl 3 . [20] The concentrations of flavonoid compounds were calculated according to the following equation that was obtained from the standard quercetin graph:

Absorbance = 0.0338 {quercetin (μg)} - 0.0002; R 2 = 0.9969

Experimental animals

Healthy adult Wistar albino rats between 5-6 month of age and weighing about (150-200 g were used for the study. The animals were housed in polypropylene cages, under standard conditions (12 h light: 12 h dark 25 ± 3°C; 35-60% humidity). All the experimental protocols were approved by Institutional Animal Ethics Committee.

Acute toxicity studies

Flavonoid-rich extract of Bauhinia variegata was studied for acute oral toxicity as per revised OECD guidelines No. 423. [21] The extract was devoid of any toxicity in rats when given in doses up to 2000 mg/kg by oral route. Hence, 200 and 400 mg/kg doses of extract were used for the study.

Nootropic activity

Elevated plus maze


Treatment groups

All the groups received the vehicle, standard drug, and plant extract one hour prior to each experiment. Overnight-fasted animals were selected and divided into groups of 5 animals each.

Group I : Control group (distilled water 10 ml/kg, per oral)

Group II : Piracetam (200 mg/kg body weight)

Group III :

Flavonoid-rich fraction of Bauhinia variegata (200 mg/kg bw)

Group IV :

Flavonoid-rich fraction of Bauhinia variegata (400 mg/kg bw).

Experimental procedure

Rats have an aversion for open-high spaces and prefer enclosed arm than the open arm. They spend more time in enclosed arm. Nootropic agents improve the acquisition process in rats subsequent to training session. Rats avoid open arm exploration and prefer enclosed arm. The plus maze has two opposite arms 50 X 10 cms, crossed with two enclosed arms of the same dimensions with walls 40 cms high. The arms were connected with a central square 10 X 10 cms, giving the apparatus a plus sign appearance. The maze was elevated at 50 cms above the floor in a dimly-lit room. Rats were placed individually in the center of the maze facing towards an enclosed arm. Initially, they were allowed to explore the maze. During the next 5 minutes, the number of entries and the time spent on the open arm and the enclosed arm were recorded. An arm entry was defined when all 4limbs were on the arm. [22]

Rotating rod activity

Treatment groups

The animals are divided into 4groups of 5 animals each.

Group I : Control group (distilled water 10 ml/kg, bw)

Group II : Diazepam (1 mg/kg bw)

Group III :

Flavonoid-rich fraction of Bauhinia variegata (200 mg/kg bw)

Group IV :

Flavonoid-rich fraction of Bauhinia variegata (400 mg/kg bw)

Experimental procedure

Rats were previously trained to remain on the rod rotating at the speed of 20 rpm for a period of 5 min. On the next day, animals are divided into 4 group 5 animal in each group. Flavonoid-rich fraction of Bauhinia variegata (200 and 400 mg/kg) and diazepam (1 mg/kg, i.p) was administered 30 min before the test, and the time required to fall off the rotating rod was noted for each animal. [23]

Effect on baclofen-induced catatonia

The animals are divided into 4 groups of 5 animals each.

Group I : Normal group

Group II : Baclofen (5 mg/kg, ip)

Group III :

Methanol Extract of Bauhinia variegata 200 mg/kg + Baclofen (5 mg/kg, ip)

Group IV :

Methanol Extract of Bauhinia variegata 400 mg/kg + Baclofen (5 mg/kg, ip)

Experimental procedure

Catatonia was measured using bar test. In brief, rats were treated with flavonoid-rich fraction of Bauhinia variegata (200 and 400 mg/kg) 30 min before Baclofen (5 mg/kg, i.p), and their forepaws were placed on the bar (0.9 cm diameter placed 2.5 cm above the table), and the duration of catatonia was measured at 5, 15, 30, 60, 90, 120 min intervals. [24]

Diazepam-induced amnesia

Treatment groups

The animals are divided into 4 groups of 5 animals each.

Group I : Control group (distilled water 10 ml/kg bw)

Group II : Diazepam (5 mg/kg bw. i.p)

Group III :

Flavonoid-rich fraction of Bauhinia variegata (200 mg/kg bw)

Group IV :

Flavonoid-rich fraction of Bauhinia variegata (400 mg/kg bw)

Experimental procedure

Four groups of rats each comprises 5 animals and weighing between 18-22 g were used. Group I was maintained as normal control, which was given with distilled water (10 ml/kg, po) once daily for 7 days on 7 th day 90 min after treatment (distilled water); transfer latency (TL) was recorded on elevated plus maze (EPM), and retention (memory) of learned task was examined 24 hr later. Group II was administered with diazepam (5 mg/kg, ip) alone on 1 st day only and after 45 min TL was recorded on EPM, and retention (memory) of learned task was examined 24 hr later.

Group III was administered with flavonoid-rich fraction of Bauhinia variegata (200 mg/kg bw) and after 45 min TL was recorded on EPM, and retention (memory) of learned task was examined 24 hr later. Group IV was administered with flavonoid-rich fraction of Bauhinia variegata (400 mg/kg bw) and after 45 min TL was recorded on EPM, and retention (memory) of learned task was examined 24 hr later. [25]

Statistical analysis

Experimental results were mean ± SEM of 6 animals. Statistical differences between the means of the various groups were evaluated using one-way analysis of variance (ANOVA) followed by Tukey test. Data were considered statistically significant only when P value < 0.05.


  Results Top


Elevated plus maze

In the plus maze test, the rats treated with flavonoid-rich fraction of Bauhinia variegata showed nootropic effect in terms of significant increase in the time spent in enclosed arm than open arm after their training sessions. This was found comparable to the standard drug and very significant as compared to the control group [Figure 1] and [Figure 2].
Figure 1: Time spent by rats after treatment with Bauhinia variegata on elevated plus maze. Significance at P < 0.05*, P < 0.01**, P < 0.001*** determined by ANOVA followed by post Hoc-Dunnet Test

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Figure 2: Number of entries by rats after treatment with Bauhinia variegata on elevated plus maze. Significance at P < 0.05*, P < 0.01**, P < 0.001*** determined by ANOVA followed by post Hoc-Dunnet Test

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Activity on rotating rod

Flavonoid-rich fraction of Bauhinia variegata (200 mg/kg) significantly decreased the fall of time to 112.5 ± 28.27 min as compared to Diazepam (1 mg/kg, ip) [Figure 3].
Figure 3: Effect of flavonoid-rich extract of Bauhinia variegata on fall-off time in mice using rota rod test. Significance at P < 0.05*, P < 0.01**, P < 0.001*** determined by ANOVA followed by post Hoc-Dunnet Test

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Effect on Baclofen-induced catatonia

Flavonoid-rich fraction of Bauhinia variegata (200 mg/kg) preponed the peak effect and at a dose of (400 mg/kg) potentiated catatonia, and peak catatonia was observed after 15 mins [Figure 4].
Figure 4: Effect of flavonoid-rich extract of Bauhinia variegata on baclofen-induced catatonia in rats. Significance at P < 0.05*, P < 0.01**, P < 0.001*** determined by ANOVA followed by post Hoc-Dunnet Test

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Diazepam-induced amnesia

Diazepam has induced dose-dependent amnesia in this amnesic model; a decrease in inflexion ratio IR was observed as compared to normal control group. Flavonoid-rich fraction of Bauhinia variegata had shown an increase in the IR and a significant reduction TL observed on EPM, and Diazepam-induced amnesia was reversed [Figure 5].
Figure 5: Effect of flavonoid-rich extract of Bauhinia variegata on diazepam-induced amnesia in rats. Significance at P < 0.05*, P < 0.01**, P < 0.001*** determined by ANOVA followed by post Hoc-Dunnet Test

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  Discussion Top


Cognitive deficits are generally observed in a number of neuropsychiatric disorders. Thus, the use of nootropics could be helpful to enhance cognitive aptitude. In the present era, exploration of novel nootropics from plants is advocated to promote cognition-associated activities. The lack/absence of major side-effects is also a reason for it. Nootropics are important groups of psychotropic drugs, which selectively facilitate protective effect on functioning of the central nervous system, basically on intellectual performance, learning capability, and memory. Nootropic drugs such as piracetam and donepezil are being used for improving memory, mood, and behavior, but as with all other psychotropic drugs, the resulting side-effects arising from them pose a major hurdle. [26],[27]

The Indian traditional system of medicine offers a number of safe treatments for central nervous system-related disorders such as anxiety and memory loss. These nature-derived treatments are effective and devoid of any untoward effects.

Flavonoids are one of the major classes of secondary metabolites that are produced by plants. Foods rich in 3 specific flavonoid sub-groups, like flavanols, anthocyanins, and/or flavanones are known to show profound potential on cognitive processes. [28],[29],[30],[31] A number of flavonoids from plants demonstrate nootropic potential. [18],[19],[32],[33],[34],[35],[36]

In the present study, Bauhinia variegata was studied for nootropic potential. Locomotor activity could be regarded as a manifestation of attentive behavior. It also indicates presence of sedative activity. Flavonoid-rich fraction of Bauhinia variegata caused no alteration in locomotion in animals. Such a response is quite expected from plants showing nootropic potential. Elevated plus maze is one of the important methods that help to assess state of anxiety in animals. It is also used to determine status of learning and memory in rats. [37] In the current study, animals treated with flavonoid-rich fraction of Bauhinia variegata (400 mg/kg) showed a significant decrease in transfer latency as compared to the control group, which indicates cognitive enhancement effect flavonoid-rich fraction of Bauhinia varigate.[38]

In the present study, effect of flavonoid-rich fraction of Bauhinia variegata was assessed by rota rod test. Rota rod test demonstrates CNS depressant activity as manifested by a decrease in fall of time. [39] In the current work, flavonoid-rich fraction of Bauhinia variegata increased fall of time as compared to diazepam as evident from statistical analysis.

Baclofen is GABA B receptor agonist, which induces catatonia in rats. Catatonia may serve as a tool for the study of GABA B receptor function. In the current work, administration of flavonoid-rich fraction of Bauhinia variegata demonstrated protective effect on rats. [40],[41] Diazepam is well-known for amnestic properties. These effects are selective for certain psychobiologically distinct memory functions. [42],[43]

Flavonoids are one of the important phytoconstituents, which could be regarded as boon in form of medicine. Flavonoids neutralize free radicals and toxic metabolites, and this compensate in loss of memory, concentration, and Alzheimer's disease. [44],[45]

Scopolamine is one of muscarinic receptor antagonists, which is known for amnestic potential, which has led to be used to study impairment of performance for various task to study integral working and reference memory. [46],[47] However, 'scopolamine reversal' is observed, because blockade in function of brain is arbitrated due to muscarinic action. In addition to this, cholinergic neurons of basal forebrain are affected in early stage of Alzheimer's disease. [48],[49]

Scopolamine is a non-selective agent for cholinergic receptor subtype. Scopolamine exerts a little effect on non-cholinergic neuronal pathway. [50] Benzodiazepines like diazepam are known to exert 'anterograde amnestic effects.' These effects are exerted due to GABA A receptor complex. This is also site for eliciting anxiolytic effects. Benzodiazepines do not blight 'impair the recall of previously learned information,' which is demonstrated by scopolamine. Thus, in the present work, 'diazepam-induced amnesia model' was followed in the study. [51]

Medhya Rasayana/Nootropic herbs are widely recognized in Ayurveda. Herbs like Centella asiatica Linn, Glycirrhiza glabra Linn, Tinospora cordifolia, Convolvulus pleuricaulis Chois, Bacopa monniera, Celastrus panniculata, Benincasa hispida, Nardostachys jatamamsi are traditionally and scientifically recognized as nootropic agents. [52]

Thus, in the present work, it could be concluded that flavonoid-rich Bauhinia variegata extract effectively demonstrated nootropic activity. However, more studies are required to validate these results.


  Acknowledgement Top


Authors are thankful to Rewa Shiksha Samiti for providing grant for this research.

 
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