Elevated Plus-maze Model Research Articles

MB‐204: A Novel Adenosine A2a Antagonist for AD, ADHD and Depression

AbstractBackgroundMB‐204 is a novel Adenosine A2a antagonist that immediately reduces depression and anxiety in animal modelsMethodMB‐204 was dosed one hour prior to its assessment for depression using the forced swim test (FST) and the elevated plus maze (EPM) for anxiety. General locomotive activity was measured as was the effect on rotarod balance as a surrogate for behavioural scores. Receptor occupancy studies were measured using radio‐labelled ligand and ex vivo brain tissue analysis.ResultMB‐204 dose dependently reduced depressive symptoms in the FST (>85%, p<0.001) with no effect on non‐specific locomotive activity or rotarod balance. MB‐204 was found to time and dose dependently occupy the A2a receptor in vivo (p<0.01) which mimicked blood PK levels. In head‐to‐head studies, MB‐204 outperformed Istradefylline (the only approved A2a antagonist for the treatment of Parkinson’s disease) in almost all measurements in the EPM model, and appeared to improve social interactions as measured by sub‐vocalization analyses.ConclusionMB‐204 appears to be an excellent pre‐clinical candidate for the treatment of A2a associated conditions

Efficacy and Anti-Inflammatory Activity of Ashwagandha Sustained-Release Formulation on Depression and Anxiety Induced by Chronic Unpredictable Stress: in vivo and in vitro Studies.

Stress is the psychological, physiological, and behavioral response of an individual's body when they perceive a lack of equilibrium between the demands placed upon them and their ability to meet those demands. Adaptogens are herbs that help with stress management, and Ashwagandha is one such safe and effective adaptogen. We evaluated the anti-neuroinflammatory potential of Ashwagandha sustained-release formulation (AshwaSR) by estimating the in vitro expression of pro-inflammatory cytokines, and its efficacy on anxiety and depression in an in vivo study. Our in vitro study investigated the anti-inflammatory potential of AshwaSR by estimating the expression of tumour necrosis factor [TNF]-α and interleukin [IL]-1β levels in LPS-induced THP-1 human monocytes, and the antioxidant effects by its potential to inhibit the superoxide [SO] generation in PMA-induced HL-60 human monocytic cells. The in vivo study assessed the efficacy of AshwaSR on chronic unpredictable stress (CUS)-induced comorbid anxiety and depression in Sprague Dawley rats. Antidepressant and anxiolytic effects of AshwaSR were evaluated by open field test (OFT), elevated plus maze (EPM), forced swim test (FST), and Morris water maze (MWM) test. AshwaSR inhibited TNF-α, IL-1β and superoxide production in a dose-dependent manner in the in vitro study. The in vivo CUS model induced depression-like and anxiety-like behaviour. Treatments with AshwaSR and escitalopram showed improvement in the EPM and MWM models compared to the CUS-group. In vitro study demonstrated that AshwaSR inhibits expressions of pro-inflammatory cytokines, IL-1β and TNF-α, and superoxide production. Further, the in vivo study confirmed its anxiolytic and stress-relieving effects in the CUS model that confirmed AshwaSR's potential in managing stress and stress-related symptoms.

Evaluation of anxiolytic effect of combination of tramadol with gabapentin and pregabalin in Wistar rats

Background: Anxiety is a widespread disorder that approximately 18% of the population experience at some stage in their lives. Pain is a common stimulus that induces anxiety in both animals and human beings. Combination of drugs with good anti-anxiety and analgesic effect can be used for the treatment of chronic pain induced anxiety, post-operative, and procedure-related pain-induced anxiety. Aims and Objectives: The study was conducted to evaluate the anxiolytic activity of combination of gabapentin with tramadol, pregabalin with tramadol compared to standard fluoxetine, in elevated plus maze (EPM) and light-dark arena (LDA) models of anxiety in Wistar albino rats. Materials and Methods: Twenty-four male or female albino Wistar rats from Central Animal House, MRMC, Kalaburagi, were utilized. Fluoxetine 10 mg/kg, gabapentin 30 mg/kg, tramadol 30 mg/kg, and pregabalin 30 mg/kg were used. Eddy’s hot plate method used for inducing anxiety. EPM and LDA models were used to study the effect of drugs in reducing the pain and anxiety. Results: The present study showed that the exposure to hot plate induces pain, creates anxiety, and reduces locomotor and explorative activity among the rats when exposed to hot plate. There was reduction in anxiety after drug administration, in fluoxetine and gabapentin with tramadol groups with >75% increase in entry into the light chamber or open arm at least once or more during the time period of 5 min when compared to hot plate post-exposure readings. Whereas in pregabalin and tramadol group, it was observed that 25% increase in entry of rats into open arm at least once during the time period of 5 min and 25% decrease in entry of rats into light chamber as compared to those rats after exposure to hot plate. Conclusion: Our study has demonstrated that tramadol, pregabalin, and gabapentin have got analgesic as well as anti-anxiety effects in rats when given in combination. All these experimental data, together with the previous experimental studies and the results reported in this work, suggest that combination of these drugs could be more effective in treating anxiety-related disorders such as chronic pain, pain-induced anxiety, post-operative, and procedure-related pain-induced anxiety with minimal side effects. Further dose ranging studies and models might be necessary to better understand the effects of these drugs in combination.

Antianxiety Activity Evaluation of Hibiscus Sabdariffa Linn. Calyces

Background: Anxiety is one of the most commonly prevailing psychiatric and behavioural disorders. Evolutionarily, it is regarded as a healthy emotion or behavioural repertoire, which is the outcome of a stressful situation, thereby, helping an individual to cope with a possible risk or a novel situation by increasing awareness and responsiveness. Benzodiazepines, barbiturates, SRIs etc., are used as first line drugs for the management of anxiety, but due to myriad of adverse effects associated with these conventional drugs, inclination of patients towards complementary and alternative medicines (CAMs) has tremendously risen. CAM interventions mainly include herbal remedies.Different parts of Hibiscus sabdariffa have been used traditionally for treating several ailments including mental disorders. Purpose: Keeping in mind the traditional use of H. sabdariffacalyces,the present study was designed to evaluate their antianxiety potential. Methods: Petroleum ether (60-80°C), chloroform, ethanol, and water extracts were prepared using successive Soxhlet extraction and evaluated using elevated plus-maze model. Results: Among all the extracts, ethanol extract showed significant antianxiety activity at a dose of 400 mg/kg. Conclusion: Thus, the study establishes antianxiety activity of H. sabdariffa calyces and also validates their traditional use.

English

Anxiety is present in several psychiatric disorders, and pharmacological treatment has limitations. Therefore, there is a search for new therapeutic approaches. This study aimed to evaluate the anxiolytic effect of acute administration of the dry extract of Passiflora alata (P. alata) leaves in male Wistar rats submitted to swimming. The animals were randomly divided into four groups (n = 8) (G1 to G4). G2 to G4 were submitted to swimming for 15 min, and 23 h later, all groups were treated via gavage: G1 (control); G2, treated with 0.9% saline solution; and G3 and G4, treated with 22 mg/kg and 66mg/kg of P. alata extract, respectively. After 1 h of administration, G2-G4 were again submitted to swimming for 5 min. All groups were submitted to the Elevated Plus Maze (EPM). The results showed that all animals submitted to swimming (G2-G4) remained less time in the open arms when compared to the control group (G1). The results demonstrate that the swimming model was adequate to induce anxiety in the animals. However, no significant differences were observed between the groups in the times of movement and immobility (in the swimming model) and the length of time in the open arms in the EPM model, indicating that in the model and doses used, P. alata extract had no anxiolytic effect. Key words: Passiflora alata extract, anxiety, swimming, wistar rats, elevated pluz maze.

Potential role of Thiocolchicoside in anxiety disorder: A pre-clinical study

Abstract Aim:The aim of the current study is to evaluate anti-anxiety and potentiating effect of Thiocolchicoside in animal models of anxiety. Methodology: A total of 24 (n=24) Swiss albino mice were procured, and they were divided into four groups of six mice in each. First group of mice (control) received 10 ml/kg-Normal Saline, second group (standard) received 2.0 mg/kg-Diazepam, test-1 received 1 mg/kg-Thiocolchicoside and test-2 received Thiocolchicoside (1mg/kg) + Diazepam (2mg/kg) for seven days per orally. All the mice were evaluated for anti-anxiety activity by Elevated Plus Maze (EPM) 60 minutes after the oral drug administration of drugs on day 1, 3 and 7 and later after a washout period of one month, same four groups of mice were screened by Light and Dark Arena (LDA) model after receiving respective drugs. Results: One-way ANOVA followed by Tukey’s Kramer test were applied for inter group comparison and correlation test for intra-group comparison. Results are expressed in mean ± SEM. In EPM, time spent in open arm for the control, standard, test- 1 and test-2 were 64.5±25.81, 128.33±17.6, 138±10.56 and 168.33±22.35 seconds respectively. There is statistically significant difference between standard (P=0.05), test-1 (P=0.05) and test-2 (P=0.03) when compared with control group. Similarly, time spent in closed arm for the control, standard, test-1 and test-2 were 238.83±20.41, 171.67± 17.6, 162±10.56 and 131.67±22.35 seconds respectively. In this, test-2 is statistically significant from the control group (P=0.05). The number of entries in the open arm for the control group, standard group, test-1 and test-2 were 3.5±1.64, 13.17±7.44, 21 ± 4.05 and 13.33 ± 2.16 respectively. There is statistically significant difference between standard (P=0.03), test-1 (P=0.02) and test-2 (P=0.03) with control. Similarly, number of entries to closed arm for the control, standard, test-1 and test-2 were 9.5±3.62, 16.33 ± 5.65, 16.33 ± 4.23 and 8.17 ± 1.72 respectively. The values obtained for the standard, test-1 and test-2 were not statistically significant (P=0.8). In LDA, time spent in light arena for the control, standard, test-1 and test-2 were 106.83±18.21, 163.5±21.66, 105.33 ± 11.57 and 125.17 ± 16.35 seconds respectively. Statistically significant difference between the control and the standard group (P=0.05) is noted. Time spent in dark arena for the control, standard, test-1 and test-2 were 193.17±18.21, 136.5±11.66, 194.67±15.57 and 174.83±16.35 seconds respectively. Here only standard group is statistically significant when compared with control (P=0.05). The number of entries in the light arena for the control, standard, test-1 and test-2 were 11.67 ± 1.37, 13.17 ± 2.48, 12 ± 2.61 and 11.67 ± 1.03 respectively. The number of entries in the dark arena for the control, standard, test-1 and test-2 were 12.17 ± 1.47, 13.17 ± 2.93, 11.83 ± 2.23 and 11 ± 0.89 respectively. With regard to number of entries in the light and dark arena there was no statistical significant difference between the groups (P>0.05). Conclusion: The study result clearly showed that Thiocolchicoside (1 mg/kg) has anti anxiety and additional potentiating effect when combined with diazepam in EPM and LDA models.

Effect of Ocimum sanctum leaf extract in animal models of anxiety

Background: Anxiety is one of the most prominent psychiatry disorders related to common stress. Approximately two-thirds of anxious patients respond to currently available treatments but the magnitude of problem is still disappointing. Ocimum sanctum (OS), a perennial shrub grown in India, has anti-stress activity. Yet, there is a paucity of data regarding this. Aims and Objectives: This study aims to evaluate the anti-anxiolytic effect of OS leaf extract (OSLE) in Swiss albino mice. Materials and Methods: Elevated plus maze (EPM) and open field test are standardized tests done for screening anxiolytic effects of drugs. The animals were grouped and the tests were conducted and the results were compared with the standard drug diazepam. Results: There was significant increase in the time spent in open arm and number of open arm entries in the diazepam group and group receiving OSLE at dose of 100 mg/kg and 200 mg/kg in EPM test. Furthermore, in the open field test, there was significant increase in the number of square crossed and rearing in the diazepam group and OSLE at dose of 100 mg/kg and 200 mg/kg. Conclusion: OSLE shows significant anxiolytic effect in EPM and open field test models in Swiss albino mice. This can be further studied to open up new possibilities for new drug development for anxiety.

In-vivo Assessment of Tranquilizer Activity of Various Extracts of Cajanus cajan Leaves in Mice

Background: Cajanus cajan is a legume of the plant family Fabaceae. Another name of this plant is the red gram, gungo pea, and no-eye pea. It is a multifaceted plant as it is widely often consumed as a dal. It contains wealthy proteins. The whole plant parts are utilized for silkworms as a vegetable; tops, green leaf, and husk are utilized as feed. Objective: The objective of the analysis was to determine the anti-anxiety effect of varied extracts viz n-hexane, chloroform, ethyl acetate, and methanol of the leaves of Cajanus cajan with an EPM (Elevated plus maze) as well as actophotometer model in albino mice. Methods: In the present research, the anti-anxiety activities of several extracts were evaluated viz n-hexane, chloroform, ethyl acetate, & methanol of the leaves of Cajanus cajan with an EPM model in albino mice. Albino mice have ministered with varied extract dosages orally (for example, 200 & 400 mg/kg) and behaviour on the EPM has been seen. The standard usage of Diazepam (2 mg/kg P.O.) (positive control). Results: Results indicate that the methanolic extract of C. cajan manifested a significant as well as maximum dose-dependent impact at 200 & 400 mg/kg on mice with the help of the EPM model and the results were exactly like diazepam (2 mg/kg), the standard antianxiety substance. The model actophotometer exhibited a dose-dependent reduction in locomotor activity compared to control animals, which is shown at two distinct dosages (200 & 400 mg/kg) of Cajanus cajan. Conclusion: The content of polyphenols was shown in the phytochemical test for methanol extract, which could be liable for the anxiolytic potential of Cajanus cajan. This plant might thus also be cultivated as a potentially beneficial anti-anxiety substance.

1H-1,2,3-triazole grafted tacrine-chalcone conjugates as potential cholinesterase inhibitors with the evaluation of their behavioral tests and oxidative stress in mice brain cells

The present paper explicates the synthesis of 1H-1,2,3-triazole tethered tacrine-chalcone conjugates and evaluation of their AChE and BuChE inhibitory activity. In-vitroAChE inhibition assay revealed three compounds, 9h, 9i, and 11f, being more potent than the standard drug tacrine and further evaluated against butyrylcholinesterase. The present study was extended to investigate the anti-amnestic effect of promising compoundson scopolamine-induced behavioral and neurochemical changes in mice. Inclined plane model and Elevated plus-maze model were performed to assess general limb motor activity and anxiety-like behavior, respectively, in mice pre-treated with scopolamine. Oxidative stress parameters reduced glutathione contents (GSH) and lipid peroxidation products (TBARS) in the brain homogenates as estimated using ex-vivo studies. Furthermore, molecular docking studies were performed for the potent compounds to decipher the mechanism of observed activities.

Potential Estrogenic Properties of 17β-Hydroxy-ethylimine Estradiol Derivative Targeted to Menopause Stage.

Hormone replacement therapy during menopause increases the risk of thromboembolic diseases and cancer, so safety alternative therapeutic strategies are needed. 17β-Aminoestrogens are a synthetic estrogens group that possess mild anticoagulant activity that contrasts with the pro-coagulant effects showed by estradiol's (E2) in rodents. Being considered an alternative to conventional hormone replacement therapy during menopause without thrombogenic risks producing. The present study aimed to determine the estrogenic profile and anxiolytic activity of 17β-[hydroxy-ethylimine]-1,3,5(10)-estratrien-3-ol (IE2), a related compound unknown until now. IE2 was assessed in immature rats by uterotrophic assay administering IE2, E2, or vehicle. In ovariectomized adult Wistar rats (Ovx) to facilitating the lordotic behavior compared with E2, estradiol benzoate, or vehicle. The effect of IE2 on anxiety was estimated in Ovx animals treated with IE2, E2, or vehicle group and evaluated in the elevated plus-maze model. IE2 produced an uterotrophic effect, lordotic behavior, and anxiolytic effect in a dose-dependent manner, similar to E2. IE2 depicted estrogenicity, indicating potential clinical use as hormone replacement therapy during menopause.

Pharmacological and In-Silico Investigations of Anxiolytic-like Effects of Phyllanthus Fraternus: A Probable Involvement of GABA-A Receptor

Background:: Phyllanthus fraternus Webster Linn (family, Euphorbiaceae) is used as a traditional medication for the treatment of various disorders and has therapeutic implications. Objective:: This study intends to investigate the anxiolytic potential of Phyllanthus fraternus standardized extract and prediction of the probable role of its marker phytoconstituents. Methods:: We tested the standardized hydro-ethanolic extract of Phyllanthus fraternus (whole plant) for the Elevated plus-maze model (EPM) and Light & Dark Exploration test as classical models for anxiety. Phyto-chemical HPTLC fingerprint analysis was performed for the detection of two classes of compounds lignans and tannins. HPTLC analysis of the standardized extract was performed using phyllanthin hypophyllanthin and corilagin as marker compounds. Additionally, GABA receptor antagonism was studied in other sets of experiments to assess the involvement of this receptor in the anxiolytic- like effects produced by Phyllanthus fraternus. Results:: The lower doses of the lignan and tannin-rich extract of the Phyllanthus fraternus possess significant anxiolytic-like activity compared to the standard diazepam. Additionally, the results of the present study suggested that high doses (400mg/kg) of Phyllanthus fraternus have exerted some sedative- like effects. Phytochemical screening and HPTLC fingerprint analysis indicate the presence of lignans and tannins, whereas HPLC analysis of the standardized extract revealed the presence of marker lignan (Hypophyllanthin) and Tannin (Corilagin). The anxiolytic-like effect of Phyllanthus fraternus observed in the mice models were blocked by Flumazenil indicating the involvement of GABAA receptors in the modulation of this effect. Our molecular docking studies also supported probable anxiolytic and sedative effects. Conclusion:: To summarize, results support the use of Phyllanthus fraternus in the anxiety-like symptoms/ disease condition and suggest its anxiolytic-like effect governed by the GABA-A receptors.

Memory Enhancing Activity of Saraswatarishta in Mice

Alzheimer's disease (AD) is a progressive neurodegenerative disorder characterized by a gradual decline in memory. Incidence of Alzheimer's disease increases with age. The disease incidence is 1% in 60 year olds & increases to 30 % at 85 years age. Hence this disease is already having enormous magnitude in today's graying world. Current treatment of Alzheimer’s disease includes- cholinesterase inhibitors& N-methyl- D-aspartate antagonists, but the benefit observed is modest. In traditional medicine Saraswatarishta is being used as memory enhancer for centuries. Brahmi, one of its major ingredients, is also being used to treat Alzheimer’s. So present study was undertaken for authentication of traditional claims of Saraswatarishta as a memory-enhancing agent. Five groups of mice (6 mice in each) were used for this study. Control group (group I) received distilled water, Group II received Saraswatarishta (2.5ml/kg) single dose and Group III received Saraswatarishta (2.5ml/kg) for 2wks. Group IV was given Diazepam (1mg/kg) to produce amnesia. For Group V, Saraswatarishta (2.5ml/kg) was given for 2wks followed by Diazepam (1mg/kg). Effect of Saraswatarishta on learning and memory of mice was studied using elevated plus maze model (EPM). Reduction in TL (Transfer Latency) indicates improvement in learning or memory and prolongation indicates impairment. Diazepam induced prolongation of TL is an accepted model of dementia. In our study, 2 weeks daily treatment of Saraswatarishta completely prevented impairment of learning and memory by Diazepam, corroborating the Ayurvdic use of Saraswatarishta and Brahmi, its major ingredient in the management of dementia. Saraswatarishta can be used as preventive measure to overcome demensia in Alzheimer’s disease.

EVALUATION OF NEUROPROTECTIVE EFFECT OF AZILSARTAN AS MEMORY ENHANCER AGAINST SCOPOLAMINE AND SODIUM NITRITE-INDUCED AMNESIA IN SWISS ALBINO MALE RATS

Objective: The objective of the study was to assess the neuroprotective effect of azilsartan as a memory enhancer against scopolamine-induced amnesia in rats.
 Methods: Albino Swiss male rats in equal numbers per group (n=6) were taken. Scopolamine hydrobromide was administered to induce amnesia within the rats. Control group rats were administered normal, negative control groups were administered with scopolamine to induce amnesia and nitrite during trials, positive control group rats were administered piracetam+ scopolamine and piracetam nitrite during trials, and test control group rats were administered azilsartan +sodium nitrite and azilsartan nitrite. Exteroceptive behavioral models just like the elevated plus-maze model, Morris water maze model, acquisition trials, and retrieval trial were wont to evaluate the neuroprotective effect of azilsartan.
 Results: The scopolamine and azilsartan have a significantly decreasing effect on time spent within the target quadrant (TSTQ) but piracetam has an increasing effect. The effect of azilsartan on transfer latency time (TLT) was observed against scopolamine-induced amnesia in rats using the elevated plus-maze test. Piracetam was found to decrease the TLT and restore memory function at a better dose. Within the case of scopolamine treated rats, a big increase in TLT was noted. Azilsartan treated group also increased TLT within the elevated plus maze. It is noted that the scopolamine features a significantly increasing effect on escape latency time (ELT). Piracetam features a decreasing effect on ELT. A rise in ELT was seen because of azilsartan.
 Conclusion: This study suggested that the azilsartan features a significant decreasing effect on TSTQ, azilsartan treated group also increased TLT and ELT.

Kaempferol Facilitated Extinction Learning in Contextual Fear Conditioned Rats via Inhibition of Fatty-Acid Amide Hydrolase.

Background: Fear, stress, and anxiety-like behaviors originate from traumatic events in life. Stress response is managed by endocannabinoids in the body by limiting the uncontrolled retrieval of aversive memories. Pharmacotherapy-modulating endocannabinoids, especially anandamide, presents a promising tool for treating anxiety disorders. Here, we investigated the effect of kaempferol, a flavonoid, in the extinction of fear related memories and associated anxiety-like behavior. Methods: The ability of kaempferol to inhibit fatty-acid amide hydrolase (FAAH, the enzyme that catabolizes anandamide) was assessed in vitro using an enzyme-linked immunosorbent assay (ELISA) kit. For animal studies (in vivo), the extinction learning was evaluated using contextual fear conditioning (CFC, a behavioral paradigm based on ability to learn and remember aversive stimuli). Furthermore, an elevated plus-maze (EPM) model was used for measuring anxiety-like behavior, while serum corticosterone served as a biochemical indicator of anxiety. Lastly, the interaction of kaempferol with FAAH enzyme was also assessed in silico (computational study). Results: Our data showed that kaempferol inhibited the FAAH enzyme with an IC50 value of 1 µM. In CFC, it reduced freezing behavior in rats. EPM data demonstrated anxiolytic activity as exhibited by enhanced number of entries and time spent in the open arm. No change in blood corticosterone levels was noted. Our computational study showed that Kaempferol interacted with the catalytic amino acids (SER241, PHE192, PHE381, and THR377) of FAAH enzyme Conclusion: Our study demonstrate that kaempferol facilitated the extinction of aversive memories along with a reduction of anxiety. The effect is mediated through the augmentation of endocannabinoids via the inhibition of FAAH enzyme.

Anxiolytic activity of Psidium guajava in mice subjected to chronic restraint stress and effect on neurotransmitters in brain.

The anxiolytic activity of Psidium guajava L. leaf ethanolic extract (PLE) and its effect on neurotransmitter systems was investigated. PLE, extracted using Soxhlet apparatus, was subjected to preliminary qualitative and quantitative (flavonoids and phenols) analyses. The anxiolytic activity at 100, 200, and 400 mg/Kg doses were assessed in mice using elevated plus maze (EPM) and light/dark transition (LDT) test models on days 1 and 16. Neurotransmitters such as monoamines (serotonin, norepinephrine, and dopamine), γ-aminobutyric acid (GABA), and glutamate were estimated in different regions of the brain (cortex, hippocampus, and cerebellum and brain stem). Phytoconstituents identified using gas chromatography-mass spectrometry and liquid chromatography-mass spectrometry were analyzed in silico to evaluate their potential binding mode(s) to GABAA and 5-HT1A receptors. Phytochemical studies showed the presence of alkaloids, tannins, flavonoids, saponins, anthraquinone glycosides, carbohydrates, and proteins, whereas total flavonoid and phenol contents were estimated to be 64.96 ± 0.95 and 206.58 ± 1.60 mg/g of dried extract, respectively. PLE treatment significantly enhanced exploratory activity of mice in EPM and LDT models with significant effects on monoamines, GABA and glutamate levels in the brain. The in silico studies suggested the interaction(s) of PLE component(s) with GABAA /5-HT1A receptors as a potential mechanism of its anxiolytic activity.

An Insight Into the Anxiolytic Effects of Lignans (Phyllanthin and Hypophyllanthin) and Tannin (Corilagin) Rich Extracts of Phyllanthus amarus : An In-Silico and In-vivo approaches.

This investigation was aimed at exploring the anxiolytic potential of Phyllanthus amarus standardized extracts and predict probable role of marker phyto constitutents. Three standardized extracts of Phyllanthus amarus plant viz. standardized aqueous extract of Phyllanthus amarus whole plant (PAAE), standardized methanolic extract of P. amarus leaf (PAME) and the standardized hydro-methanolic extract of P. amarus leaf (PAHME) were tested in the classical animal models of anxiety: Elevated plus-maze model and Light & Dark Exploration test. The lower doses of the tannin rich extract (PAHME) of the P. amarus possess significant anxiolytic activity compared to lignin rich (PAME) and aqueous extracts (PAAE), while at a higher dose (400mg/kg) the results of all three extracts appears to be potentially sedative. While the molecular docking studies support these probable anxiolytic, the sedative effects of the Phyllanthus amarus extracts could be due to the interaction of tannins and lignans with the GABAbenzodiazepine receptor complex. The results of the present study indicate that the tannin-rich extract of the P. amarus may have potential clinical applications in the management of anxiety. It can be further studied for optimum dosage to be used as a future of anti-anxiety drug development or as a standardized Phytomedicine.

Preclinical Evaluation of Anxiolytic Activity of Adhatoda Vasica Leaf Extract in Swiss Albino Mice

Objectives: To evaluate the possible anxiolytic activity of methanolic extract of Adhatoda vasica leaves (MEAVL) in Swiss albino mice. Methods: A total of 60 (n=60) mice were used. They were divided into ten groups of six mice each. The dose of 200, 400 and 600 mg/kg MEAVL were chosen after acute toxicity study. First 5 groups were evaluated by Elevated Plus Maze (EPM) and rest 5 group were evaluated by Light and Dark Arena (LDA) model. First group of mice (control) received 0.05ml/10g of Normal Saline, second group (standard) received 1.0 mg/kg Diazepam, third, fourth and fifth group received 200, 400 and 600 mg/kg of MEAVL respectively. Anti-anxiety effect was evaluated 60 min after the oral drug administration. Results: All the results were expressed in mean ± SD. One way ANOVA followed by Tukey- Kramer multiple comparison tests were used to compare between groups. In EPM model, the numbers of entries in open arm were 3.50±1.64, 13.17±7.44, 7.50±4.63, 3.67±1.86 and 4.67±2.94 respectively noted for 1st to 5th groups respectively for Normal Saline, Diazepam, 200, 400 and 600 mg/kg of MEAVL. Similarly in LDA model, the numbers of entries in dark arena were 12.17±1.47, 13.17±2.92, 12.33±3.01, 7.17±1.94 and 10.50±3.08 respectively noted for 6th to 10th group. This result clearly showed that MEAVL-400mg/KG has antianxiety property when compared to diazepam as evidenced by increase in the number of entries in dark arena in LDA model with p 0.05). Conclusion: The current study has demonstrated an antianxiety effect of MEAVL in a dose of 400mg/kg in LDA model. However, the study plant must be further evaluated to confirm the molecular basis of the antianxiety property so that it can be helpful for the society in the near future because of its antianxiety property.

Nanostructured lipid carriers-mediated brain delivery of carbamazepine for improved in vivo anticonvulsant and anxiolytic activity.

The limited brain delivery of carbamezapine (CBZ) presents a major hurdle in the successful epilepsy treatment. The potential of carbamezapine-loaded nanostructured lipid carriers (CBZ-NLCs) for improved brain delivery is investigated in the current study. CBZ-NLCs were prepared by using binary mixture of trilaurin and oleic acid as a lipid core stabilized with Poloxamer 188, Tween 80 and Span 80. CBZ-NLCs were evaluated for physicochemical properties, in vitro release, in vivo brain kinetics, anticonvulsant and anxiolytic activities. The optimized CBZ-NLCs demonstrated nanometric particle size (97.7nm), surface charge of -22mV and high drug incorporation (85%). CBZ-NLCs displayed biphasic release pattern with initial fast followed by sustained drug release. CBZ-NLCs significantly enhanced the AUC of CBZ (520.4µg·h/mL) in brain compared with CBZ dispersion (244.9µg·h/mL). In vivo anticonvulsant activity of CBZ-NLCs in PTZ-induced seizure model showed a significant increase in the onset time (143.0 sec) and reduction in duration (17.2 sec) of tonic-clonic seizures compared with CBZ dispersion (75.4 and 37.2 sec). The anxiolytic activity in light-dark box and elevated-plus maze models also demonstrated superiority of CBZ-NLCs to CBZ dispersion. From the results, CBZ-NLCs presents a promising strategy to improve brain delivery and therapeutic outcomes of CBZ in epilepsy.

Effect of Oral Honey Administration on Sleep-Deprived Male Mice

Background: Sleep is a biologic process that is essential for life and optimal health. Sleep plays a critical role in brain function and systemic physiology. The deleterious health consequences of sleep deprivation are associated with risks for a wide variety of medical conditions. Honey’s potential role in restorative sleep may have implications in improving long term health. Objective: To determine the effect of honey on sleep deprivation in male mice. Method: The study was carried out using four (4) groups of young male mice (N=5-6 per group) deprived of sleep for a period of 6 hours. Bee honey was administered orally at three dose levels; 10 %, 20 % and 40 % V/v respectively to three groups while the control group was administered normal saline (vehicle). Novelty-Induced Behaviour (NIB), Elevated plus-maze (EPM), Hole board and Y-maze models were used to evaluate the effects of honey on the mice. Results: In the NIB model, a significant (p<0.05) decrease in locomotion was observed dose-dependently. The same observation was recorded for rearing behaviour while a biphasic effect was observed in grooming behaviour. The results obtained in the other models showed that locomotor activity was significantly decreased suggesting that honey has central inhibitory effect. In the hole board test and EPM, there were significantly (p<0.05) decreased activity of the mice due to the administration of honey. Conclusion: This study demonstrates that honey exerts dose-dependent central inhibitory effects in sleep-deprived male mice therefore suggesting possible amelioration of sleep deprivation effects.

Lawsonia Inermis Markedly Improves Cognitive Functions in Animal Models and Modulate Oxidative Stress Markers in the Brain.

Background and Objective: Medicinal plants represent an important source of alternative medicine for the management of various diseases. The present study was undertaken to assess the potential of Lawsonia inermis ethanol (Li.Et) and chloroform (Li.Chf) extracts as memory-enhancing agents in experimental animals. Materials and Methods: Li.Et and Li.Chf were phytochemically characterized via gas chromatography-mass spectroscopy (GC-MS). Samples were tested for nootropic potentials at doses of 25, 50, 100, 200 mg/kg (per oral in experimental animals (p.o.)). Swiss albino mice of either sex (n = 210) were divided into 21 × 10 groups for each animal model. Memory-enhancing potentials of the samples were assessed using two methods including “without inducing amnesia” and “induction of amnesia” by administration of diazepam (1 mg/kg, intraperitoneally. Piracetam at 400 mg/kg (i.p.) was used as positive control. Cognitive behavioral models including elevated plus maze (EPM) and the passive shock avoidance (PSA) paradigm were used. Biochemical markers of oxidative stress such as glutathione (GSH), catalase (CAT), superoxide dismutase (SOD) levels were analyzed in the brain tissue of treated mice. Results: In 2,2-diphenyl-1-picrylhydrazyl (DPPH) free radicals scavenging assay, Li.Et and Li.Chf exhibited 70.98 ± 1.56 and 66.99 ± 1.76% inhibitions respectively at 1.28 mg/mL concentration. GCMS results revealed the presence of important phytochemicals. Both samples (Li.Et and Li.Chf) at 25 mg/kg (p.o.) dose significantly (p < 0.05) improved learning and memory as indicated by decline in transfer latency and increase in step down latency in EPM and PSA models respectively. Li.Et and Li.Chf at 25 mg/kg (p.o.) showed considerable increase in GSH (2.75 ± 0.018 ***), SOD (2.61 ± 0.059 ***) and CAT (2.71 ± 0.049 ***) levels as compared to positive and negative control groups. Conclusions: This study provides the preliminary clue that L. inermis may be a potential source of memory-enhancing and anti-oxidant compounds and thus warrant further studies.