|Year : 2018 | Volume
| Issue : 3 | Page : 141-147
Comparative Efficacy of Ekapākī (1x) and Daśapākī (10x) Brāhmī Ghr̥ta in Chronic Unpredictable Mild Stress Induced Behavioral Alterations in Rats
Swati S Gadgil1, Asmita A Wele2, Vidushi R Sharma1, Madhuri S Pawar1
1 Department of Rasashastra Evam Bhaishajyakalpana, College of Ayurved, BharatiVidyapeeth (Deemed to be University), Katraj, Pune, Maharashtra, India
2 Department of Pharmacology and Pharmacotherapy, University of Debrecen, Debrecen, Hungary
|Date of Submission||03-Dec-2018|
|Date of Decision||31-Jul-2019|
|Date of Acceptance||03-Aug-2019|
|Date of Web Publication||10-Feb-2020|
Dr. Swati S Gadgil
Department of Rasashastra Bhaishajyakalpana, College of Ayurved, Bharati Vidyapeeth (Deemed to be University), Katraj, Pune - 411 043, Maharashtra
Source of Support: None, Conflict of Interest: None
Context: With a rising need for development of drugs that are effective with multifaceted therapeutics for the treatment of depression and the associated behavioral changes, brāhmī ghr̥ta offers a choice of treatment. The challenge of palatability of ghr̥ta in therapeutic dose of 20 to 40 g per day can be overcome by potentiation and the same was tested in the present study. Aims: To evaluate and compare the efficacy of 1x Brāhmī Ghr̥ta (BG) and 10x BG in Chronic unpredictable mild stress (CUMS) induced behavioral alterations. Settings and Design: Randomized Controlled preclinical trial. Materials and Methods: Self prepared and standardized 1x Brāhmī ghr̥ta (3.6 g/kg) and 10x BG (3.6, 1.8 and 0.9 g/kg) were evaluated in CUMS induced behavioral changes in rats by using sucrose preference (SPT), forced swim (FST), and memory assessment with elevated plus maze (EPM) tests. Statistical Analysis Used: One-way ANOVA followed by Post hoc analysis. Results: 28 days CUMS procedure induced depression in rats which was confirmed. Upon comparison with CUMS control group, 1x BG and 10x BG, at all three dose levels, increased the sucrose preference (P < 0.05); in forced swim test increased the number of rotations (P < 0.001) and in elevated plus maze test significantly lowered the transfer latency time in retention transfer latency [1X BG (P < 0.05) and10X BG (P < 0.001)]. Further, 10x BG at all 3 dose levels showed significantly better results in comparison to 1x BG in FST (P < 0.01) and EPM (P < 0.05). Conclusions: Both the samples of brāhmī ghr̥ta āēapākī (1x) and daśapākī (10x), showed antidepressant effect and reversal of memory impairment induced by CUMS model. This is the first study to report that drug potentiation increases the efficacy and achieves dose reduction.
Keywords: Brāhmī, chronic unpredictable mild stress, potentiation, Siddha Ghr̥ta
|How to cite this article:|
Gadgil SS, Wele AA, Sharma VR, Pawar MS. Comparative Efficacy of Ekapākī (1x) and Daśapākī (10x) Brāhmī Ghr̥ta in Chronic Unpredictable Mild Stress Induced Behavioral Alterations in Rats. Ancient Sci Life 2018;37:141-7
|How to cite this URL:|
Gadgil SS, Wele AA, Sharma VR, Pawar MS. Comparative Efficacy of Ekapākī (1x) and Daśapākī (10x) Brāhmī Ghr̥ta in Chronic Unpredictable Mild Stress Induced Behavioral Alterations in Rats. Ancient Sci Life [serial online] 2018 [cited 2020 Oct 29];37:141-7. Available from: https://www.ancientscienceoflife.org/text.asp?2018/37/3/141/277985
| Introduction|| |
Poor drug compliance of medicated ghee or oil prescribed in neuropsychological disorders poses major challenge to ayurveda physicians. The present study on Siddha ghr̥ta (medicated ghee) of two potencies using well known antidepressant Bacopa monnieri (Brāhmī) as a single drug was conducted to trace the change and subsequent effect.
Brāhmī (Bacopa monnieri Linn) is one among the most praised mēdhya rasāyana drugs–class of herb taken to sharpen intellect and attenuate mental deficits. It was initially described in Atharva-vēda and also in the treaties like Caraka saṣhitā and Suśruta saṣhitā.Brāhmī is attributed with properties like life-span and cognition enhancer, voice quality, luster and complexion promoter, anti-aging etc., Owing to its high potential especially in the domain of various neuropsychological conditions it has been extensively researched in CDRI laboratory.
Brāhmī is well known for its anti-oxidant, hepatoprotective, neuroprotective, anti-depressant, anxiolytic, immunostimulatory, anti-inflamatory and anticonvulsant potentials in pre-clinical trials., Current evidence suggests Bacopa monnieri acts via the following mechanisms antioxidant neuroprotection (via redox and enzyme induction), acetylcholinesterase inhibition and/or choline acetyltransferase activation, b-amyloid reduction, increased cerebral blood flow, and neurotransmitter modulation (acetylcholine [ACh], 5-hydroxytryptamine [5-HT], dopamine [DA]). Clinically brāhmī is known for safely enhancing the cognitive performance in elderly and in individuals with already high cognitive abilities. It improves the cognitive function in cerebro-vascular accident and cerebral palsy patients with behavioral problems. It is perhaps the most extensively, in depth researched herb which is also unambiguous.
In the Chronic Unpredictable Mild Stress (CUMS) animal model of depression, the animals are exposed to continuous mild stress using various stressors such as food deprivation, water deprivation, constant illumination, tilted cages, and soiled cages in a random unpredictable manner. Usually, after two weeks of CUMS, the animals develop variety of symptoms similar to depressive symptoms in human beings such as weight loss, altered diurnal rhythms, sleep disturbances, and anhedonia. Unlike other stress models, the CUMS mimics the ways humans encounter stressors on daily basis and thus is more suitable for elucidation of pathological impacts of stress. Preclinical and clinical studies have shown that prolonged stress plays a prominent role in memory deteriorations and represents a major risk factor that triggers alzheimer's disease pathology. The effects of stress on memory include interference with the individual's ability to encode memory and retrieve vital information.
Several disadvantages are associated with the current pharmacotherapy for depression. For example, it takes around 4–6 weeks' time for the onset of antidepressant effect and approximately 30% of patients remain unresponsive to currently available drugs. Also, several side effects limit the patient compliance to therapy. Thus there is a need to have a drug with a better efficacy with multifaceted actions and that could target various aspects of the underlying pathologies of the depression.
Siddha ghr̥ta is a novel and unique dosage form in Ayurved wherein active constituents from herbs, minerals or animal products are extracted into the lipids. It is of foremost importance when neuropsychological conditions are involved. The base drug cow ghee possesses mēdhya (cognitive enhancer) activity and when it is processed with a drug like brāhmī its activity multiplies. Siddha ghr̥ta exhibitspoor drug compliance owing to its high dose of about 20 to 40 g per day, greasy nature, strong smell and taste. To combat these issues; āvartana, which literally means repetition of the procedure can prove helpful. Thus present work was carried out for evaluation and comparison of efficacy of standardized brāhmī ghr̥ta with 1x potency (1x BG) (prepared with a single processing cycle - ēkapākī) and standardized brāhmī ghr̥ta with 10x potency (10x BG) (prepared with ten processing cycles - daśapākī) using CUMS induced behavioral changes gauged by sucrose preference test, forced swim test and elevated plus maze test in rats.
| Materials and Methods|| |
1x BG and 10x BG were prepared using only two ingredients i.e. cow ghee and brāhmī juice extracted from fresh brāhmī plant. Drugs were prepared by complying with the guidelines in ayurvedic treaties and were standardized using Bacoside A as a marker compound.
Adult wistar rats of either sex (180-250 g) were procured from CPCSEA approved Central animal house, BVDU Medical College, Pune. They were housed under standard (23-300 C, 50–60% humidity) laboratory conditions, maintained on a 12-h natural day–night cycle. Unless otherwise stated; animals had free access to standard food and water. They were acclimatized to laboratory conditions before onset of the experiment. Approval was obtained from Institutional Animal Ethics Committee (Ref: BVDUMC/2228/2017/001/001).
This experiment was conducted by using the simultaneous drug induction and challenge method. Here 1X BG was used in therapeutic dose while 10X BG was used in therapeutic; half and also one fourth of the therapeutic dose with an intention to check whether or not potentiation results in drug dose reduction.
Forty two rats (six in each group) were randomly divided in to seven groups. Group I consisted of unstressed rats (normal control), Group II comprised of stressed rats (disease control), group III consisted of stressed rats receiving imipramine hydrochloride (10mg/kg) (positive control) (trade name Tab. Depranil 25 mg of Manas Pharma diluted in distilled water to prepare a desired concentration), group IV consisted of stressed rats receiving 1x BG (3.6 g/kg) (therapeutic dose equivalent to 40 g in Humans), group V, VI and VII consisted of stressed animals receiving 10x BG in the dose of 3.6 g/kg therapeutic dose, 1.8 g/kg half of the therapeutic dose and 0.9 g/kg one fourth of the therapeutic dose. Study drugs were liquefied using water bath just before dosing; and administered. Unstressed rats were housed together and were kept in a different room than the rest of the groups. All the drugs were administered orally; daily for twenty eight days one hour before exposure to stressor.
Chronic unpredictable mild stress procedure
Rats were exposed to various stress schedules as confirmed by previous researchers with minor modifications., Animals were subjected to one of the stress schedule in random order once a day for 28 days. Following Stressors were used: 1hour separation in an empty bottle, 24 hours without bedding, high density housing (12 or more rats in one cage) for 24 h, housing in soiled cage for 24 h, overnight illumination for 12 h, 30 second tail pinch, 450 cage tilt for 24 h, food and water deprivation for 24h, 60 second tail pinch, cold water (4°C) swim for 5 min.
Body weight measurement
Body weight of each animal was measured every day and recorded on 1st, 4th, 8th, 11th, 15th, 18th, 22nd, 25th, and 28th day. Rate of change in weight over the 28 days duration was calculated by following formula: Rate of change in weight = (Final wt. - Baseline wt.)/Duration of treatment in days.
Sucrose preference test
Sucrose preference test (SPT) was done as described previously by Kumar et al. 2011 with minor modifications to test anheadonia-related behavior. It consisted of training and testing courses. During training; rats were deprived of food and water for 48 hr. prior to exposure to 1% sucrose solution. For next three days; routine food and water supply was provided. On 6th day they were again deprived of food and water for next 23 hrs starting at 9:00 am and on day seven SPT was performed between 8:00-9:00 am to record baseline readings. This time rats were exposed to two pre-weighed bottles kept in each cage, one bottle of 1% sucrose solution and another containing normal tap water. Rats had a choice of selection in between these two bottles. The consumed quantity was recorded. Test was repeated in the same manner on day thirty six. Sucrose preference was calculated using the following formula: Sucrose Preference = Sucrose intake (g)/ [Sucrose intake (g) + Water intake (g)] × 100.
Forced swim test
It was performed the next day after SPT. A water tank (30 × 20 × 15 cm) with a rotating wheel (25 cm diameter) at its centre was used. The tank was filled with water up to height of 13 cm maintained at 25 ± 10 C. Each rat was placed in the tank and the door was locked so that rat could not escape out. In an effort to escape out of water, rat tries to swim and end up with rotating the wheel. Thus, each rat was observed for five minute duration and the rotations displayed on the digital counter were recorded. After each procedure, the water in the tank was changed with water of similar temperature. In FST; decreased number of rotations is suggestive of depressive behavior.
Elevated plus maze test
It was performed on the next day after FST as stated by Rinwa and Kumar 2014 with minor modification. Animals were subjected to acquisition and retention trial using elevated plus maze. The apparatus consists of four arms, two closed arms measuring 50 cm × 40 cm × 10 cm and two open arms of the dimensions 50 cm × 10 cm. From central platform the arms extended up to 10 cm × 10cm and the plus maze is elevated to a height of 50 cm from the ground. In elevated plus maze transfer latency on first day was considered as initial transfer latency. The retention/consolidation (memory) was examined 24 h later and considered as retention transfer latency.
All the rats were given training on the first day i.e. on 38th day, each rat was placed at the distal end of an open arm, facing away from the central platform and the time taken by rat to move into one of the closed arms with all its four legs, was recorded i.e. Transfer latency (TL). If the animal did not enter into one of the closed arms within 90s, he was gently pushed into one of the two closed arms and the TL was assigned as 90s. The rat was allowed to explore the maze for another 90s. After 24hr i.e. on day thirty-nine; retention of this learned-task i.e. memory was recorded using same methodology by recording TL. Experiment was conducted in a dimly lit, semi soundproof room under natural light. [Figure 1] shows the total study schedule from day one up to day thirty eight.
Data was expressed as Mean ± standard deviation. Statistical analysis was carried out using Graphpad Instat software version 3 and differences were considered significant at P < 0.05. The normal control group was compared with the CUMS control group. The drug treated animals were compared with the CUMS control and with each other also. The data was analyzed using one-way analysis of variance (ANOVA) followed by post hoc test.
| Results|| |
Effect of chronic unpredictable mild stress and drug treatment on body weight
Body weights were assessed to confirm the efficacy of CUMS. Rate of change in weight over the duration of 28 days was calculated and compared as shown in [Table 1]. One-way ANOVA revealed that there was a significant difference in the groups. Post hoc analysis (Kruskal Wallis test) indicated a decrease in the rate of body weight gain of the animals exposed to CUMS treatment compared to normal control animals (P < 0.001) over the period of 28 days; chronic pre-treatment with 1x BG (3.6 g/kg) and imipramine significantly (P < 0.05) increased the body weight over the duration of 28 days compared to CUMS group. However there was no significant difference between 10x BG (3.6 g/kg, 1.8 g/kg and 0.9 g/kg) groups and CUMS group with regard to weight gain (P > 0.05).
|Table 1: Effect of chronic unpredictable mild stress and drug treatment on body weight (g) over a period of 28 days duration|
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Effect of chronic unpredictable mild stress and drug treatment on sucrose preference test
[Figure 2] shows the effect of CUMS and CUMS along with drug treatment on sucrose preference (%) after 28 days duration. The reduced consumption of the sucrose solution is the indicator of anhedonia-like behavior. In the initial baseline test, one way ANOVA revealed that there is no significant (P > 0.05) differences between various treatment groups. Data is not shown here. Data (after 28 days treatment) was analyzed using one way analysis of variance followed by post hoc Tukey-Kramer multiple comparisons test. 28 days exposure to CUMS significantly (* P < 0.05) reduced the percentage of sucrose preference when compared with the non stressed group (normal control). However administration of imipramine and various study drugs for the period of 28 days demonstrated significant (# P < 0.05) percentage increase for sucrose preference compared to CUMS group. There was no significant alteration in percentage of sucrose preference ((P > 0.05) between rats in 1x BG and 10x BG (three dose levels) groups.
|Figure 2: Effect of chronic unpredictable mild stress and drug treatment on sucrose preference (%) after 28 days duration. N = 6/group. All values represent mean ± SD; *P < 0.05 compared to Normal control group,# P < 0.05 compared to CUMS control (DC) group SD: Standard deviation, NC: Normal control, Disease Control (DC): CUMS control, PC: Positive control|
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Effect of chronic unpredictable mild stress and drug treatment on forced swim test
[Figure 3] shows effect of CUMS and drug treatment on forced swim test. Data was analyzed using one way analysis of variance followed by post hoc Tukey-Kramer multiple comparisons test. Number of rotations of CUMS group (DC) was significantly (P < 0.001) decreased when compared with non stressed normal control group. Treatment with 1x BG (3.6 g/kg), imipramine and 10x BG (3.6, 1.8 and 0.9 g/kg) at all three dose levels significantly (P < 0.001) increased the number of rotations as compared to CUMS group. Treatment with 10x BG (3.6, 1.8 and 0.9 g/kg) at all three dose levels significantly (P < 0.001) increased the number of rotations compared with treatment with 1x BG (3.6 g/kg). Treatment with imipramine also showed increase in the number of rotations (P < 0.05) when compared with the treatment with 1x BG (3.6 g/kg).
|Figure 3: Effect of chronic unpredictable mild stress and drug treatment on forced swim test. N = 6/group. All values represent mean ± SD; ***P < 0.001 as compared to normal control and###P < 0.001 as compared to disease control,αP < 0.05,ααP < 0.01 as compared to 1x BG. SD: Standard deviation, NC: Normal control, Disease Control (DC): CUMS Control, PC: Positive control|
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Effect of chronic unpredictable mild stress and drug treatment on elevated plus maze test
[Figure 4] demonstrates the results on EPM test. Data was analyzed using one way analysis of variance followed by post hoc Tukey-Kramer multiple comparisons test. After 28 days exposure to CUMS and drug treatment, there was no significant difference (P > 0.05) in first day latency (initial transfer latency) (ITL) among the groups. Data is not presented here. However, there was a significant difference between groups in the retention transfer latency (RTL). TL of CUMS group (DC) was significantly (P < 0.001) increased when compared with non stressed normal control group demonstrating stress-induced memory impairment. Treatment with 1x BG (3.6 g/kg) and imipramine significantly (P < 0.05) lowered TL in RTL as compared to CUMS group. Treatment with 10x BG (3.6, 1.8 and 0.9 g/kg) also significantly (P < 0.001) lowered the TL in RTL compared to CUMS group. Treatment with 10x BG (3.6, 1.8 and 0.9 g/kg) significantly (P < 0.05) lowered the TL in RTL compared to treatment with 1x BG (3.6 g/kg) group. Treatment with 10x BG (3.6 g/kg) significantly (P < 0.05) lowered TL in RTL compared with treatment with imipramine. Treatment with 10x BG (1.8 and 0.9 g/kg) significantly (P < 0.01) lowered TL in RTL compared with treatment with imipramine. Treatment with 1x BG (3.6 g/kg) and imipramine showed significantly (P > 0.05) higher TL in RTL compared to normal control group demonstrating that both drugs were not able to protect memory impairment due to CUMS comparable to non stressed normal group.
|Figure 4: Effect of chronic unpredictable mild stress and drug treatment on elevated plus maze test on initial transfer latency and retention transfer latency. N = 6/group. All values represent mean ± SD; *P < 0.05 and ***P < 0.001 as compared to normal control and#P < 0.05,###P < 0.001 as compared to disease control,αP < 0.05 as compared to 1x BG,+P < 0.05,++P < 0.01 as compared to positive control|
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| Discussion|| |
Objective of the present study is to evaluate and compare efficacy of 1x BG and 10x BG (used at three dose levels) using CUMS induced behavioral changes. The chronic administration of various uncontrollable stressors in an unpredictable manner is a well-documented animal model for the preclinical evaluation of antidepressants. CUMS is generally thought to be the most promising and valuable rodent model to study depression in animals, mimicking several human depressive symptoms. It imitates stressful situations of peoples' everyday life. Impulses that initiate the stress response in laboratory animals, so-called stressors, act usually from two to four weeks, and are potentially harmless to the body. Furthermore, long-term exposure (of experimental animals) to various mild stressors is associated with significant change in the behaviors. Thus in the present study depression was induced by CUMS and subsequent behavioral alterations were assessed using SPT, FST and EPM test.
The CUMS procedure (28 days) displayed a significant decrease in the rate of body weight gain in stressed rats when compared to non stressed rats as shown in the [Table 1]. Chronic treatment with 1x BG and imipramine could be able to attenuate this decrease in the rate of body weight gain when compared with CUMS control. 1x BG is ghee based formulation which was prepared using juice of brāhmī with a single processing cycle and 10x BG was prepared using juice of brāhmī (in each cycle) with ten processing cycles. Reddy M et al. 2013 have reported the percentage increase in the body weight of ghee fed rats was 32.8 in a period of forty days; which is quite high. In agreement with this finding we can say that 1x BG containing greater concentration of lipids than the concentration of brāhmī could be responsible for little more body weight gain in 28 days duration. 10x BG has higher concentration of brāhmī as compared to concentration of lipids. Also it was heated for 31 days. Heating (agnisaṣskāra) is known to make the product lighter (laghu). A light in nature substance does not increase body weight, is a well known Ayurveda principle. That's why it's found that 1X BG shows slight increase in body weight but not 10X BG.
The sucrose preference test represents the anhedonia-like behavioral change. Anhedonia was modelled by inducing a decrease in responsiveness to rewards reflected by a reduced consumption of sucrose solutions; it is a core symptom of human major depression. In this study, CUMS exposed rats (after 28 days schedule) showed a reduced preference of sucrose solution as compared to non-stressed control group animals. This result is in agreement with the previous findings. Treatment with 1x BG, 10x BG (at all three dose levels) and imipramine administered in parallel to CUMS procedure significantly reversed this behavioral change; which represents the antidepressant-like effect of the study drugs in CUMS induced depression.
Forced swim test is the behavioral despair test and had been frequently used to determine depressant-like behavior in rodents after exposure to stress. The immobility time or decreased number of rotations of FST reflects 'behavioral despair' as seen in human depression. In this study; CUMS rats showed decreased number of rotations as compared to non stressed rats indicating depressive–like behavior. All the treatment drugs showed positive effect with increase in the number of rotations compared to CUMS group suggestive of reversal of the depressive behavior.
Elevated plus maze test was used to assess effect of CUMS on memory. Many studies have reported memory deficits in depressed subjects. An effect size analysis of cognitive functioning in 726 patients with major depressive disorder (MDD), conducted using meta-analytic principles, found that the type of memory task most affected by depression was recollection. Recollection memory is analogous to explicit or declarative memory, requiring the conscious recall of specific facts or events. Present experiment showed increase in the transfer latency time in the RTL in rats exposed to CUMS when compared with non-stressed rats indicating probable deficit in the recall memory, in agreement with the above fact. Both the study drugs along with the standard drug imipramine displayed decreased TL in the RTL when compared with CUMS treated group; indicating reversal of memory impairment due to CUMS procedure.
Clinical and pre-clinical data show that depression is associated with activation of the immune system, which is manifested as the inflammation state. Moreover, in the CNS, the microglia plays an important role in the synapses; signalling and neurotransmission control and also systematically regulate the normal functions of the immune system. It is a proven fact that bacopa inhibits the release of inflammatory cytokines from microglial cells and inhibits enzymes associated with the inflammation in the brain limiting the inflammation of the CNS. This anti-inflammatory activity of bacopa can be responsible for positive results of the study drugs in reversing the behavioral changes due to CUMS.
Recent clinical studies of depression have paid special attention to the hippocampus and frontal cortex, which are brain regions structurally and functionally affected by stress responses and critically involved in the regulation of mood and learning/memory function. Brain derived neurotrophic factor (BDNF) is a member of the nerve growth factor family and is essential for growth, maintenance, cellular differentiation and survival of neurons in the central nervous system. It modulates neuronal plasticity, inhibits cell death cascades and increases the cell survival proteins that are responsible for the proliferation and maintenance of central nervous system neurons; hence, it may be an important factor in the development and treatment of depression. A study demonstrated that chronic unpredictable stress exposure decreased BDNF protein and mRNA levels in both the hippocampus and frontal cortex of depressed rats and long term treatment with Bacopa monnieri (80-120 mg/kg) significantly reversed the CUS induced changes in behavior, BDNF protein and mRNA levels. Consistent with this view, we can postulate that 1x BG and 10x BG at all three dose levels have displayed their positive effect on CUMS induced depression and memory assessed using SPT, FST and EPM, probably by up regulating the BNDF proteins in the hippocampus decreased due to exposure to CUMS.
Antioxidants can produce improvement in cognitive function by protecting neurons against the injurious effect of reactive oxygen species. Oxidative stress occurs in brain tissues whenever there is increased generation of reactive oxygen species and impaired antioxidant defense mechanism. A study demonstrated that the activities of antioxidant enzymes like SOD, CAT and GSH significantly diminished in the hippocampus; and MDA (lipid peroxidation marker) level was found elevated after exposure to chronic unpredictable stress. Chronic administration of Bacopa monnieri extract (80mg/kg) for five consecutive weeks increased the activities of SOD and CAT, increased the levels of GSH, inhibited lipid peroxidation and reduced the production of MDA in chronic unpredictable stress. Thus by virtue of antioxidant and neuroprotective effect of brāhmī which must have been present in 1x BG and 10x BG, both might have protected neurons against oxidative stress-mediated injury and prevented stress induced depression and memory loss.
In FST and EPM test, post-hoc test indicated a significant difference in the results of 1x BG and 10x BG (at all three dose levels). 10x BG at all three dose levels showed better results in both the behavioral tests when compared with 1x BG group. The major chemical entity shown responsible for neuropharmacological effects of Bacopa monniera is bacoside A (64.28%) and bacoside B (27.11%). 10x BG is prepared using brāhmī juice repeatedly for ten times and hence contain higher concentration of phytochemical from brāhmī including bacoside A in comparison with 1x BG. Thus this higher concentration must be responsible for better results of 10x BG at half (1.8 g/kg) and one-fourth (0.9 g/kg) of the therapeutic dose, which is actually the purpose of potentiating the drug.
It is evident that orally taken Bacopa monnieri extract (BME) was up taken into the system by showing presence of bioactive compound bacoside A in the serum of BME treated rats. The bioactive compounds in the BME could directly or indirectly interact with neurotransmitter systems to enhance learning and memory. Since the bacosides present in the BME are nonpolar glycosides, they can cross the blood-brain barrier (BBB) by simple lipid-mediated passive diffusion. To add to this effect; both the study drugs are lipoid in nature and have a natural affinity towards brain tissue facilitating the easy entry of the drug into the tissue.
To sum up with the observations found; it can be said that 1x BG and 10x BG at all three dose levels demonstrated reversal of all behavioral changes induced due to CUMS. Additionally 10x BG being a potentiated version; demonstrated significant results at half and one-fourth of the therapeutic dose as compared to 1x BG. This study is the first and foremost attempt to generate evidence between the relationship of drug potentiation and drug dose reduction. Further extended research is required to find out exact mechanism of action of the drug; as well its efficacy in reduced dose needs to be clinically evaluated.
| Conclusions|| |
1x BG (3.6 g/kg) and 10x BG (3.6 g/kg, 1.8 g/kg and 0.9 g/kg) showed antidepressant effect and reversal of memory impairment induced by CUMS model. The study established that drug potentiation increases the efficacy of the drug and thus drug dose can be reduced and drug can be dispensed in a better and palatable form like soft gel capsules.
Source of Support
Authors are thankful to Dr. Vijaya Pandit, Head, Department of Pharmacology, Medical College, Bharati Vidyapeeth (Deemed to be University), Katraj, Pune for making the animals and animal house facilities available. Authors are also thankful to Dr. Kanchan Borole and Dr. Jayashri Dawane for their guidance and help during the animal study conducted.
Conflicts of interest
There are no conflicts of interest.
| References|| |
Aguiar S, Borowski T. Neuropharmacological review of the nootropic herb Bacopa monnieri
. Rejuvenation Res 2013;16:313-26.
Singh HK. Brain enhancing ingredients from āyurvedic medicine: Quintessential example of Bacopa monniera
, a narrative review. Nutrients 2013;5:478-97.
Mathur D, Goyal K, Koul V, Anand A. The molecular links of re-emerging therapy: A review of evidence of brahmi (Bacopa monniera
). Front Pharmacol 2016;7:44.
Calabrese C, Gregory WL, Leo M, Kraemer D, Bone K, Oken B. Effects of a standardized Bacopa monnieri
extract on cognitive performance, anxiety, and depression in the elderly: A randomized, double-blind, placebo-controlled trial. J Altern Complement Med 2008;14:707-13.
Kumar N, Abichandani LG, Thawani V, Gharpure KJ, Naidu MU, Venkat Ramana G. Efficacy of standardized extract of Bacopa monnieri
(Bacognize®) on cognitive functions of medical students: A six-week, randomized placebo-controlled trial. Evid Based Complement Alternat Med 2016;2016:4103423.
Ramakrishnan K, Sathya, Sajgotra E. Efficacy of brahmi in cognitive function – A comparison between cerebral palsy children and C.V.A patients. Int J Innov Res Sci Eng Technol 2017;6:6945-52.
Abdul Shukkoor MS, Baharuldin MT, Mat Jais AM, Mohamad Moklas MA, Fakurazi S. Antidepressant-like effect of lipid extract of Channa striatus
in chronic unpredictable mild stress model of depression in rats. Evid Based Complement Alternat Med 2016;2016:2986090.
Umukoro S, Omorogbe O, Aluko O, Eduviere T, Owoeye O, Oluwole O. Jobelyn®, a sorghum-based nutritional supplement attenuates unpredictable chronic mild stress-induced memory deficits in mice. J Behav Brain Sci 2015;5:586-97.
Gadgil S, Wele A. Drug potentiation with subsequent extraction of biomarker: A case study of brāhmīghr̥ta. J Res Educ Indian Med 2018;24:71-80.
Kumar B, Kuhad A, Chopra K. Neuropsychopharmacological effect of sesamol in unpredictable chronic mild stress model of depression: Behavioral and biochemical evidences. Psychopharmacology (Berl) 2011;214:819-28.
Zhang L, Luo J, Zhang M, Yao W, Ma X, Yu SY. Effects of curcumin on chronic, unpredictable, mild, stress-induced depressive-like behaviour and structural plasticity in the lateral amygdala of rats. Int J Neuropsychopharmacol 2014;17:793-806.
Shang X, Shang Y, Fu J, Zhang T. Nicotine significantly improves chronic stress-induced impairments of cognition and synaptic plasticity in mice. Mol Neurobiol 2017;54:4644-58.
Gupte PA, Dawane J, Wele AA. Experimental evaluation of Hiṅgvādi Ghṛta in behavioral despair using animal models. Anc Sci Life 2016;36:84-9.
Rinwa P, Kumar A. Unpredictable, mild, stress-induced depressive-like behaviour and structural plasticity in the lateral amygdala of rats. Naunyn Schmiedebergs Arch Pharmacol 2014;387:129-41.
Parle M, Singh N. Animal models for testing memory. Asia Pac J Pharmacol 2004;16:101-20.
Gumuslu E, Mutlu O, Sunnetci D, Ulak G, Celikyurt IK, Cine N, et al.
The antidepressant agomelatine improves memory deterioration and upregulates CREB and BDNF gene expression levels in unpredictable chronic mild stress (UCMS)-exposed mice. Drug Target Insights 2014;8:11-21.
Pekala K, Budzynska B, Biala G. Utility of the chronic unpredictable mild stress model in research on new antidepressants. Curr Issues Pharm Med Sci 2014;27:97-101.
Reddy M, Satish V, Nagamounica M, Nagamounica M, Kumar M. Myths and facts about consumption of ghee in relation to heart problems – A comparative research study. Int J Pharm Pharm Sci 2013;5:560-3.
Tianzhu Z, Shihai Y, Juan D. Antidepressant-like effects of cordycepin in a mice model of chronic unpredictable mild stress. Evid Based Complement Alternat Med 2014;2014:438506.
Dong H, Gao Z, Rong H, Jin M, Zhang X. B-asarone reverses chronic unpredictable mild stress-induced depression-like behavior and promotes hippocampal neurogenesis in rats. Molecules 2014;19:5634-49.
Xi G, Hui J, Zhang Z, Liu S, Zhang X, Teng G, et al.
Learning and memory alterations are associated with hippocampal N-acetylaspartate in a rat model of depression as measured by 1H-MRS. PLoS One 2011;6:e28686.
Reus GZ, Abelaira HM, Leffa DD, Quevedo J. Cognitive dysfunction in depression: Lessons learned from animal models. CNS Neurol Disord Drug Targets 2014;13:1860-70.
Nemetchek MD, Stierle AA, Stierle DB, Lurie DI. The ayurvedic plant Bacopa monnieri
inhibits inflammatory pathways in the brain. J Ethnopharmacol 2017;197:92-100.
Banerjee R, Hazra S, Ghosh AK, Mondal AC. Chronic administration of Bacopa monniera
increases BDNF protein and mRNA expressions: A study in chronic unpredictable stress induced animal model of depression. Psychiatry Investig 2014;11:297-306.
Kumar S, Mondal AC. Neuroprotective, neurotrophic and anti-oxidative role of Bacopa monnieri
on CUS induced model of depression in rat. Neurochem Res 2016;41:3083-94.
Rajan KE, Preethi J, Singh HK. Molecular and functional characterization of Bacopa monniera
: A retrospective review. Evid Based Complement Alternat Med 2015;2015:945217.
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