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1.  Evaluation of Anxiolytic-Like Effect of Aqueous Extract of Asparagus Stem in Mice 
There are few studies on the neuropharmacological properties of asparagus, which was applied in Chinese traditional medicine as a tonic and heat-clearing agent. The present study was designed to investigate the anxiolytic-like activity of the aqueous extract of asparagus stem (AEAS) using elevated plus maze (EPM) and Vogel conflict tests (VCT) in mice. AEAS significantly increased the percentage of time spent in open arms in EPM, when compared with control group. In the Vogel conflict drinking test, the numbers of punished licks increased to 177% and 174% by the treatment of AEAS at the doses of 1.5 and 3.0 g/kg (250 and 500 mg sarsasapogenin per kilogram of body weight), compared with control group. The serum cortisol level decreased significantly, at the same time. In conclusion, these findings indicated that the aqueous extract of asparagus stem exhibited a strong anxiolytic-like effect at dose of 1.5 and 3.0 g/kg (250 and 500 mg sarsasapogenin per kilogram of body weight) in experimental models of anxiety and may be considered an alternative approach for the management of anxiety disorder.
PMCID: PMC3853311  PMID: 24348707
2.  Chemical composition and anxiolytic evaluation of Achillea Wilhelmsii C. Koch essential oil in rat 
Herbal based remedies are used worldwide to treat psychiatric disorders. The aim of this study was to analyse the essential oil composition of Achillea Wilhemsii C. Koch (Asteraceae) and to evaluate its anxiolytic effects in the elevated plus maze (EPM) model of anxiety in rat. Gas chromatography/mass spectrometry (GC/MS) analysis of the essential oil showed that the main compounds of the oil were p-ocimen (23%), 1, 8-cineole (20.8%) and carvone (19.13%). The EPM results showed that 1 mg/kg (i.p.) of the oil significantly (P<0.05) increased the percentage of the time spent and the number of entries in the open arms of the maze while it did not change the total number of entries in the maze arms. These effects were not reversed with 2 mg/kg flumazenil and 5 mg/kg naloxone. We concluded that a minimum dose of 1 mg/kg of the oil has anxiolytic effects which are not probably mediated through GABA and opioid receptors.
PMCID: PMC3757592  PMID: 24082896
Achillea Wilhelmsii C. Koch; Anxiolytic; Essential oil; GC/MS; Elevated plus maze
3.  Effect of Cissampelos Pareira Leaves on Anxiety-like Behavior in Experimental Animals 
The present study was undertaken to evaluate anxiolytic effect of 70% hydroethanolic extract of leaves of Cissampelos pareira in murine models. C. pareira (Menispermaceae) is rich in alkaloids, and phytochemical results showed that it contains alkaloids, flavanoids, terpenoids, steroids, etc., Anxiolytic activity was evaluated by using elevated plus maze test (EPM), light dark (LandD) model, and forced swim test (FS) models in rats. The efficacy of extract (100, 200, 400 mg/kg) was compared with control as well as standard diazepam (DZ; 2 mg/kg, p.o.) in EPM, LandD model, and imipramine (IM; 2.5 mg/kg, p.o.) in FS model. The results showed that DZ and extract significantly increased the number of entries, time spent in open arm, head dip counts, and rearing time, while they decreased fecal count in EPM. DZ and extract also significantly increased the number of crossings and time spent in light compartment, while they decreased duration of immobility in LandD model. In case of FS model, IM and extract significantly increased mobility and swimming time. Thus, the results confirm that hydroethanolic extract of C. pareira has the potential to be used in the management of anxiety-like behavior in a dose of 200 and 400 mg/kg. Further study is required to explore the plant and its parts for anxiolytic potential.
PMCID: PMC3924988  PMID: 24716177
Anxiolytic; Cissampelos pareira; Elevated plus maze; Forced swim test; Light dark model
4.  Role of Monoaminergic System in the Etiology of Olive Oil Induced Antidepressant and Anxiolytic Effects in Rats 
ISRN Pharmacology  2013;2013:615685.
Olive oil is the major component of the Mediterranean diet and has rich history of nutritional and medicinal uses. In the present study, the antidepressant and anxiolytic effects and their neurochemical basis following repeated administration of extravirgin olive oil were monitored. Male albino Wistar rats were used during study. Animals of test group were given olive oil orally at the dose of 0.25 mL/kg daily for 4 weeks. Control rats received equal volume of water. Elevated-plus maze (EPM) test and forced swim test (FST) were performed for the assessment of anxiety and depression like symptoms. An increase in time spent in open arm in EPM and increased struggling time in FST following long-term administration of olive oil indicate that olive oil has anxiolytic and antidepressant properties. Neurochemical results showed that repeated administration of olive oil decreased the levels of brain 5-HT (5-hydroxytryptamine), 5-HIAA (5-hydroxyindoleacetic acid), and levels of DA (dopamine); however, levels of DA metabolite HVA (homovalinic acid) were increased. Hence, present findings suggest that olive oil has neuroprotective effects. It reduces behavioral deficits via altering 5-HT and DA metabolism. So it could be used as a therapeutic substance for the treatment of depression and anxiety.
PMCID: PMC3725699  PMID: 23936669
5.  Anti-anxiety Activity of Methanolic Extracts of Different Parts of Angelica archangelica Linn. 
Angelica archangelica a herb distributed in tropical and subtropical regions of the world. In Indian and Chinese system of medicine, it is used for nervous disorders and cerebral diseases. Previously the aqueous extract of the A. archangelica was evaluated for anxiolytic activity and was found to have significant potential for the same. The present study is aimed to evaluate the anxiolytic activity of methanol extract of root (MER), stem (MES), leaf (MEL), fruit (MEF) and whole plant (MEW) of Angelica archangelica Linn. All the extracts (MER, MES, MEL, MEF and MEW) were evaluated for anxiolytic effects using elevated plus maze test (EPM) model in rats. Methanol extracts of different parts of A.archangelica had increased number of entries and time spent in open arms while they decreased the number of entries and duration of time spent in closed arm of the EPM. In a similar fashion, the diazepam increased the percentage of time spent and percentage of arm entries in the open arms (*P <0.05, **P <0.01). Whole plant and the root had the maximum, leaf and fruits showed intermediate, while stem had the least anxiolytic activity (*P <0.05, **P <0.01) in EPM (Figure 1-5). The head dip count in DZ, SMR400, SML400, SMF400 and SMW400 in open arm are significantly shown in Table 1. The DZ, SMF400 and SMW did not show the fecal bolus while other groups were reduced the fecal bolus significantly (**P <0.01) as compared to control (Table 1). Whole plant and leaf showed the most, root and fruit the intermediate and stem the least anxiolytic activity (**P <0.01) in EPM.
PMCID: PMC3942901
Angelica archangelica; Anxiolytic; Elevated plus maze test
6.  The Ontogeny of Anxiety-Like Behavior in Rats from Adolescence to Adulthood 
Developmental Psychobiology  2010;52(8):731-739.
In human beings, susceptibility to anxiety disorders can be relatively high during adolescence. Understanding the ontogeny of anxiety-like behavior in laboratory rodents has implications for developing anxiolytic drugs that are suitable for this age group. Given the dearth of information about adolescent rodents, this study examined the response of both male and female adolescent, late adolescent, young adult, and older adult rats to three tests of anxiety-like behavior: the emergence test (ET), open field (OF), and elevated plus-maze (EPM). The results showed that adolescent rats exhibited a higher anxiety-like response than adults on each test; the amount of locomotion in the OF and percentage of time spent on the open arms of the EPM increased across the age groups, while older adult rats made the fewest start box re-entries in the ET. These results support the hypothesis that adolescent rats have a more pronounced response to stressors than do adults. © 2010 Wiley Periodicals, Inc. Dev Psychobiol 52: 731–739, 2010.
PMCID: PMC3061011  PMID: 21117243
adolescence; anxiety; rats; open field; elevated plus-maze; sex differences
7.  A Novel Elevated Plus-Maze Procedure to Avoid the One-Trial Tolerance Problem 
The elevated plus-maze (EPM) test is one of the most commonly used behavioral assays to evaluate anxiety-related behavior in rodents. It is an economic test (5 min duration) without prior conditioning of the animals. The critical measure for anxiety is the time spent in the open arms of the maze. A confounding problem of the EPM is the so called one-trial tolerance (OTT), characterized by a marked decrease of open arm exploration in spite of treatment with anxiolytic acting benzodiazepines upon re-exposure to the EPM. This consistent finding is often raised as an evidence for the inappropriateness to re-test rodents in the EPM. However, a reliable re-test paradigm would broaden the usability and effectiveness of this test. Therefore, we tested how an extension of the inter-trial interval to 28 days (instead of the usual 24 h), and an additional change of the testing room would affect the open arm time and other behaviors on the EPM. In two experiments, drug-naive Wistar rats were exposed to the EPM on trial 1, and treated intraperitoneally with either vehicle or midazolam (0.25 mg/kg) 30 min before trial 2. Then, trial 2 (28 days after trial 1) was carried out in either the same testing room (Experiment 1) or in another unfamiliar room (Experiment 2). Twenty-eight days after trial 1 the open arm time of the rats in the vehicle treated control rats of both experimental groups was comparable to that of the first trial, independent of the testing room. Most importantly, we found that the treatment with the benzodiazepine midazolam had a significantly anxiolytic-like (i.e., increase of open arm time) effect in trial 2 only when conducted in the previously unfamiliar testing room (Experiment 2). We suggest that in order to reliably re-test the EPM and to prevent confounding effects due to the OTT, an inter-trial interval of 28 days and a change in testing rooms reinstates anxiolytic-like actions of benzodiazepines.
PMCID: PMC3146044  PMID: 21845176
re-test EPM; one-trial tolerance; midazolam; motivation; memory
8.  Anxiolytic activity of methanol leaf extract of Achyranthes aspera Linn in mice using experimental models of anxiety 
Indian Journal of Pharmacology  2012;44(1):63-67.
To study the anxiolytic activity of methanol extract of Achyranthes aspera Linn (Amaranthaceae).
Materials and Methods:
Male Swiss albino mice were used. Methanolic extract of Achyranthes aspera (MEAA) was administered in the doses of 100, 300 and 600 mg/kg p.o. Hole board (HB), open field (OF), elevated plus maze (EPM) and light/dark exploration (LDE) tests were used for determination of anxiolytic activity.
The methanolic extract of Achyranthes aspera significantly increased the number and duration of head poking in HB test. The extract also significantly increased the time spent and the number of entries in open arm in EPM. In LDE test, the extract produced significant increase in time spent and number of crossings and decreased the duration of immobility in light box. In OFT, the extract showed significant increase in number of rearing, assisted rearing and the squares crossed.
In the present study, MEAA exhibited anxiolytic activity which might be attributed to its phyto-constituents viz. alkaloid, steroid and triterpenes. Since Achyranthes aspera is ubiquitous and abundantly grown, it could be a fairly economical therapeutic agent for management of anxiety disorders.
PMCID: PMC3271542  PMID: 22345872
Achyranthesaspera; anxiolytic activity; elevated plus maze; hole board; light/dark exploration; MEAA; open field
9.  Evaluation of Behavioral and Pharmacological Effects of Hydroalcoholic Extract of Valeriana prionophylla Standl. from Guatemala 
There are few studies on the pharmacological properties of Valeriana prionophylla Standl. (VP), known as “Valeriana del monte”, and used in Mesoamerican folk medicine to treat sleep disorders. This study examines the pharmacological effects of the hydroalcoholic extract of the dry rhizome using the open field, rota rod, elevated plus-maze (EPM), forced swimming (FST), strychnine- and pentobarbital-induced sleeping time, PTZ-induced seizures, and the inhibitory avoidance tests. VP did not show any protective effect against PTZ-induced convulsions. In the EPM, exhibited an anxiolytic-like effect through the effective enhancement of the entries (38.5%) and time spent (44.7%) in the open arms, when compared with control group. Time spent and the numbers of entrances into the enclosed arms were decreased, similar to those effects observed with diazepam. In the FST, acute treatment with VP, produced a dose-dependent decrease in immobility time, similarly to imipramine. VP also produced a significant dose-dependent decrease in the latency of sleeping time, while producing an increase in total duration of sleep; influenced memory consolidation of the animals only at lower doses, unlike those that produced anti-depressant and anxiolytic effects. In summary, the results suggest that VP presents several psychopharmacological activities, including anxiolytic, antidepressant, and hypno-sedative effects.
PMCID: PMC3132466  PMID: 21754942
10.  Anxiolytic and Anticonvulsant Effects on Mice of Flavonoids, Linalool, and α-Tocopherol Presents in the Extract of Leaves of Cissus sicyoides L. (Vitaceae) 
The aim of the present study is to demonstrate the anxiolytic and anticonvulsant effects of a hydroalcoholic extract obtained from the aerial parts of Cissus sicyoides L. (CS) (Vitaceae) on male and female mice using several behavioral assays. Groups of males and females treated via intraperitoneal (IP) with doses of 300, 600, and 1000 mg/kg of the extract showed significant action in the elevated plus-maze (EPM), time spent in the open arms, and number of entries in the open arms. The board-hole test also showed a significant increase in the time spent in head-dipping and in marble-burying test of the number of marbles buried. The same treatment increased the duration of sleeping time induced by sodium pentobarbital and also showed a significant increase in protection against pentylenotetrazole-induced convulsions. These results indicate an anxiolytic and anticonvulsant-like action from C. sicyoides L. extract on mice, probably due to the action of flavonoid(s), Linalool, and α-tocopherol present in the C. sicyoides leaves.
PMCID: PMC2655362  PMID: 19300520
11.  Anti-anxiety and anti-depressant activities of Sarasvata choorna in experimental animals 
Ayu  2011;32(4):590-593.
The present study was undertaken to evaluate the anxiolytic and anti-depressant activity of Sarasvata choorna. The anxiolytic activity was evaluated in elevated plus maze (EPM) and the anti-depressant activity was evaluated in forced swimming test (FST). The efficacy of Sarasvata choorna was compared with the standard anti-anxiety (diazepam 2 mg/kg) and anti-depressant (imipramine – 5 mg/kg) drugs. It was observed that Sarasvata choorna at the dose of 390 mg/kg is as effective as standard drugs used in anti-anxiety and anti-depressant activities in mice by increasing time spent in open arm and entries to open arm in EPM model and increasing immobility time in FST model respectively. Hence it can be concluded that Sarasvata choorna may be used as a potent therapeutic agent in treating anxiety and depressive disorders.
PMCID: PMC3361941  PMID: 22661860
Anxiolytic; anti-depressant; elevated plus maze; imipramine; Sarasvata choorna
12.  Evaluation of the anxiolytic effect of Nepeta persica Boiss. in mice 
The aim of the present study was to evaluate the anxiolytic effects of hydroalcoholic extract (HE) of Nepeta persica Boiss. (Lamiaceae) on the elevated plus-maze (EPM) model of anxiety. The extract of arial parts of the plant was administered intraperitoneally to male NMRI mice, at various doses, 30 min before behavioural evaluation. The HE extract of N. persica at the dose of 50 mg kg−1 significantly increased the percentage of time spent and percentage of arm entries in the open arms of the EPM. This dose of plant extract affected neither animal's locomotor activity nor ketamine-induced sleeping time. The 50 mg kg−1 dose of the plant extract seemed to be the optimal dose in producing the anxiolytic effects, lower or higher doses of the plant produce either sedative or stimulant effects. At 100 mg kg−1, the plant extract increased the locomotor activity. These results suggested that the extract of N. persica at dose of 50 mg kg−1 possess anxiolytic effect with less sedative and hypnotic effects than that of diazepam and causes a non-specific stimulation at 100 mg kg−1.
PMCID: PMC2396471  PMID: 18604252
anxiety; elevated plus-maze; Nepeta persica; sedative
13.  Malathion induces anxiety in the male adult mouse 
The cholinergic nervous system and acetylcholine esterase are involved in chronic intoxication with organophosphorous insecticides. The present study aims to investigate the influence of the chronic toxicity of these chemicals on behaviors related to anxiety, using the elevated plus maze (EPM), in the male adult mouse.
Material and methods
Either water or 1% concentration of malathion was applied dermally to the male adult mice (10 s, once daily for 28 days) and, on day 29, the EPM test was done.
Time spent in the open arms (TSOA) in intoxicated animals was decreased by over 50% compared to the controls (p = 0.047). In contrast, time spent in closed arms was significantly higher in the malathion-exposed mice (p = 0.025). Percentage of open arm entries (OAE) was slightly smaller in the malathion-treated group in comparison to the control animals. Percentage of closed arm entries (CAE) in the treated group was slightly higher than the value in the control animals.
The results showed that chronic toxicity of malathion may lead to an anxiety-like behavior in the animal model used in this study. It is difficult to extend these findings to clinical situations. However, more experimental work in different animal species as well as epidemiological studies in human subjects in this regard are highly recommended.
PMCID: PMC3648820  PMID: 23671451
anxiety; elevated plus maze; malathion; mouse; organophosphates
14.  Hypoxic-ischemic injury decreases anxiety-like behavior in rats when associated with loss of tyrosine-hydroxylase immunoreactive neurons of the substantia nigra 
Neonatal Sprague-Dawley rats were randomly divided into normal control, mild hypoxia-ischemia (HI), and severe HI groups (N = 10 in each group at each time) on postnatal day 7 (P7) to study the effect of mild and severe HI on anxiety-like behavior and the expression of tyrosine hydroxylase (TH) in the substantia nigra (SN). The mild and severe HI groups were exposed to hypoxia (8% O2/92% N2) for 90 and 150 min, respectively. The elevated plus-maze (EPM) test was performed to assess anxiety-like behavior by measuring time spent in the open arms (OAT) and OAT%, and immunohistochemistry was used to determine the expression of TH in the SN at P14, P21, and P28. OAT and OAT% in the EPM were significantly increased in both the mild (1.88-, 1.99-, and 2.04-fold, and 1.94-, 1.51-, and 1.46-fold) and severe HI groups (1.69-, 1.68-, and 1.87-fold, and 1.83-, 1.43-, and 1.39-fold, respectively; P < 0.05). The percent of TH-positive cells occupying the SN area was significantly and similarly decreased in both the mild (17.7, 40.2, and 47.2%) and severe HI groups (16.3, 32.2, and 43.8%, respectively; P < 0.05). The decrease in the number of TH-positive cells in the SN and the level of protein expression were closely associated (Pearson correlation analysis: r = 0.991, P = 0.000 in the mild HI group and r = 0.974, P = 0.000 in the severe HI group) with the impaired anxiety-like behaviors. We conclude that neonatal HI results in decreased anxiety-like behavior during the juvenile period of Sprague-Dawley rats, which is associated with the decreased activity of TH in the SN. The impairment of anxiety and the expression of TH are not likely to be dependent on the severity of HI.
PMCID: PMC3854134  PMID: 22147192
Anxiety; Tyrosine hydroxylase; Substantia nigra; Hypoxia-ischemia
15.  Anxiolytic-Like Effects of Chrysanthemum indicum Aqueous Extract in Mice: Possible Involvement of GABAA Receptors and 5-HT1A Receptors 
Biomolecules & Therapeutics  2012;20(4):413-417.
Chrysanthemum indicum Linne is an ancient herbal medicine used to treat bone and muscle deterioration, ocular infl ammation, headache, and anxiety in Korea, China, and Japan. Furthermore, tea derived from Chrysanthemum indicum Linne has been used to treat anxiety by facilitating relaxation and curing insomnia. However, no reports exist on the anxiolytic-like effects of Chrysanthemum indicum Linne water extract (CWE) in mice. In the present study, we investigated the anxiolytic-like effects of CWE using the elevated plus-maze (EPM) test in mice. CWE, at a dose of 500 mg/kg (p.o.), signifi cantly increased the time spent in the open arms of the EPM compared to a vehicle-injected control group. Moreover, the effect of CWE (500 mg/kg) was blocked by bicuculline (a selective GABAA receptor antagonist) and WAY 100635 (a selective 5-HT1A receptor antagonist). Taken together, these fi ndings suggest that the anxiolytic-like effects of CWE might be mediated by the GABAA receptor and the 5-HT1A receptor.
PMCID: PMC3762266  PMID: 24009829
Chrysanthemum indicum Linne; Anxiolytic-like effects; Elevated plus maze; GABAA receptor; 5-HT1A receptor
16.  Effects of Ocimum sanctum and Camellia sinensis on stress-induced anxiety and depression in male albino Rattus norvegicus 
Indian Journal of Pharmacology  2010;42(5):283-288.
The aim of this study was to study the ameliorative effects of Ocimum sanctum and Camellia sinensis on stress-induced anxiety and depression.
Materials and Methods:
The study was carried out using male albino rats (200 ± 50 g). The effect of O. sanctum and C. sinensis was evaluated for anxiety and depression using elevated plus maze (EPM) test, open field test (OFT), forced swim test (FST), and tail suspension test (TST).
Restraint stress (3 h/day for six consecutive days) induced a significant reduction in both the percentage number of entries and time spent in open arms in EPM, and these changes were reversed with post-treatment of aqueous extract of O. sanctum and C. sinensis (100 mg/kg for 6 days). Restraint stress-induced (a) increased latency and (b) decreased ambulation and rearing were also reversed by O. sanctum and C. sinensis in OFT. A significant increase in immobility period was observed in FST and TST after restraint stress. O. sanctum and C. sinensis significantly reduced the immobility times of rats in FST and TST.
O. sanctum and C. sinensis possess anxiolytic and antidepressant activities.
PMCID: PMC2959210  PMID: 21206619
Anxiety; Camellia sinensis; depression; Ocimum sanctum; restraint stress
17.  Anxiolytic and antidepressant-like effects of Melissa officinalis (lemon balm) extract in rats: Influence of administration and gender 
Indian Journal of Pharmacology  2012;44(2):189-192.
To analyse the behavioral effects of Melissa officinalis extract in rats following acute or subacute treatment.
Materials and Methods:
The behavioral effects of an acute or subacute (10-day course) orally administered M. officinalis (MO; 0, 30, 100 or 300 mg/kg) ethanol extract were evaluated in male and female Wistar rats in elevated plus-maze (EPM), forced swimming (FS) and open field (OF) tests. The effects of diazepam (DZP; 1 mg/kg) and fluoxetine (FXT; 10 mg/kg) were also assessed.
In the EPM test, the percentage of open arm entries and open arm times of both males and females given the subacute M. officinalis ethanol extract were significantly higher than those of the vehicle-treated animals but were at levels similar to those observed in the DZP group, regardless of the treatment length. In the FS test, immobility duration was significantly lower in both males and females treated with the plant extract when compared to vehicle-treated counterparts. A 10-day treatment with FXT induced the same antidepressant response, regardless of gender, and was more effective than the M. officinalis extract. Male and female rats demonstrated distinct gender profiles, and treatment × gender interactions were observed. Locomotion in the EPM and OF tests was not significantly altered by treatments.
The potential psychoactive properties of M. officinalis may provide a unique pharmacological alternative for certain psychiatric disorders; however, the efficacy appears to be dependent on both gender and administration length.
PMCID: PMC3326910  PMID: 22529473
Anxiety; depression; gender; locomotion; Melissa officinalis
18.  Evaluation of Anxiolytic effect of Erythrina mysorensis Gamb. in mice 
Indian Journal of Pharmacology  2012;44(4):489-492.
Aim and Objectives:
To evaluate anxiolytic effect of stem bark ethanol and chloroform extracts of Erythrina mysorensis in mice.
Materials and Methods:
The anxiolytic activity was examined by using the elevated plus maze (EPM) and open field test (OFT), and motor coordination by rotarod test (RRT). Twenty four Swiss albino male mice were divided into four groups of six mice each. Group 1 received vehicle (normal saline); group 2 received diazepam (1 mg/kg); groups 3 and 4 received ethanolic and chloroform extract of Erythrina mysorensis, 200 and 400 mg/kg p.o., respectively.
Mice treated with diazepam (1 mg/kg, p.o.) showed significant (P < 0.001) increase ini the percentage of open arms entries and time spent whereas, in closed arm the number of entries and time spent were significantly (P < 0.05) decreased. Oral administration of chloroform and ethanol extract of E. mysorensis exhibited significant (P < 0.05) increase in the number of open arm entries and time spent with significant (P < 0.05) reduction in number of entries and time spent in the closed arm as compared to group 1. Chloroform and ethanol extracts treated mice also produced significant increase in the number of rearings (P < 0.05), assisted rearings and number of squares crossed (P < 0.01). Rotarod test showed significant (P < 0.01) reduction in motor activity at 45 min with diazepam and E. mysorensis extracts (400 mg/kg) as compared to groups 3 and 1.
Erythrina mysorensis possess significant anxiolytic activity in the mice. It can be a promising anxiolytic agent.
PMCID: PMC3469953  PMID: 23087511
Elevated plus maze test; Erythrina mysorensis; open field test; Rotarod test
19.  Sildenafil enhances locomotor activity in young mice and exerts anxiogenic effects in both young and aged mice 
Sildenafil is a selective PDE5 inhibitor that increases cGMP levels in the target tissues and is an effective treatment agent for erectile dysfunction. The nitric oxide-cGMP pathway might be implicated in regulation of certain CNS functions, including locomotor activity and anxiety.
The aim of the current study was to investigate effects of sildenafil (3 and 10 mg/kg) on anxiety and locomotor activity in open field and elevated plus maze (EPM) tests in young and aged mice.
Sildenafil (3 and 10 mg/kg) significantly decreased the percent of time spent in the open arms compared to the control group in young animals in the EPM test, but only the 10 mg/kg dose significantly decreased the percentage of total number of entries to the open arms in young animals. Sildenafil (3 and 10 mg/kg) significantly decreased the percentage of total number of entries to the open arms in aged animals in the EPM test, but it significantly increased total distance moved and speed of the animals in the locomotor activity test in young animals. The total distance moved and the speed of the animals significantly decreased in aged animals compared to the young control group, although sildenafil (3 and 10 mg/kg) did not alter these parameters in aged mice.
Our results show that sildenafil had anxiogenic effects in young as well as aged mice, but it enhanced locomotor activity only in the young mice in the EPM test. Thus, sildenafil seems to exert different effects on anxiety and locomotion in young and aged animals.
PMCID: PMC3936918  PMID: 24500039
Sildenafil; locomotor activity; anxiety; mice
20.  Nociceptive and Anxiety-Like Behavior in Reproductively Competent and Reproductively Senescent Middle-Aged Rats 
Gender medicine  2009;6(Suppl 2):235-246.
Changes in levels of estradiol and progesterone that occur with the transition to reproductive senescence may influence nociception or affect.
To ascertain whether nociceptive and affective processes change with reproductive senescence, this study examined pain and anxiety-like behaviors in middle-aged female rats that were reproductively competent, transitioning to reproductive senescence, or reproductively senescent.
Middle-aged (12–14 months old) female rats (N = 46) were tested in the following tasks to assess pain and anxiety-like behavior: tail flick, elevated plus maze, elevated zero maze, mirror maze, Vogel punished drinking, and defensive burying. For the tail-flick task, the latency for rats to move their tail from a heat source, as an indication of pain sensitivity, was determined. In the elevated plus and elevated zero mazes, the time spent on the open arms or quadrants, respectively, were determined as measures of reduced anxiety behavior. In the mirror maze, the time spent in the mirrored portion of the chamber was used as an indicator of anxiety-like responding. In the Vogel task, the number of punished licks made was determined as a measure of reduced anxiety-like behavior. In the defensive burying task, the duration spent by rats burying an electrified prod postfootshock was utilized as an index of anxiety-like responding. All rats were experimentally naive, retired breeders from our colony and had not had a litter or been lactating for 1 to 4 weeks before behavioral testing.
Although tail-flick latencies were not significantly different among rats that were reproductively competent or senescent, reproductively competent rats had less anxiety-like behavior in the elevated plus maze (more time spent on the open arms: F2,43 = 5.93; P < 0.01), elevated zero maze (more time spent on the open quadrants: F2,43 = 4.62; P = 0.01), and Vogel punished drinking task (more punished licks made: F2,43 = 3.76; P = 0.03). There were no statistically significant differences in the mirror maze and defensive burying task.
In this study of adult female rats, nociceptive behavior did not vary significantly with reproductive senescence, but anxiety-like behavior of rats did.
PMCID: PMC2860272  PMID: 19406372
estrogen; progesterone; anxiety; affect; aging; menopause
21.  Anxiolytic property of hydro-alcohol extract of Lactuca sativa and its effect on behavioral activities of mice 
Journal of Biomedical Research  2012;27(1):37-42.
Lactuca sativa, belonging to the Asteraceae family, is a leafy vegetable known for its medicinal properties. This study aimed to understand the mechanism of Lactuca sativa extract with respect to pharmacological action.We investigated the anxiolytic effects of hydro-alcoholic extract of leaves of Lactuca sativa on mice. The behavioral tests performed on mice models to assess anti-anxiety properties were: open field test (OFT), elevated plus maze test (EPM), elevated T maze test, and marble burying test. Increased locomotor activity and time spent in the “open-arm” were observed in extract fed group. Malondialdehyde (MDA) and nitrite levels were decreased, catalase and glutathione levels were increased in Lactuca sativa treated mice. The data obtained in the present study suggests that the extract of Lactuca sativa can afford significant protection against anxiolytic activity.
PMCID: PMC3596753  PMID: 23554792
Lactuca sativa; anxiety; nitrite; open field test (OFT); elevated plus maze test (EPM); malondialdehyde (MDA)
22.  Enhanced Anxiety Observed in Cocaine Withdrawn Rats Is Associated with Altered Reactivity of the Dorsomedial Prefrontal Cortex 
PLoS ONE  2012;7(8):e43535.
Discontinuation of drug intake in cocaine abusers commonly produces a variety of adverse withdrawal symptoms among which anxiety and depression-related behavior are prevailing during the initial period of abstinence. The aim of this study was to provide further insight into the neurobiological dysregulations that might contribute to these pathological states. Rats were treated with cocaine or saline for 14 days (20 mg/kg; i.p) and anxiety-related behavior was assessed in different paradigms (elevated plus-maze (EPM), confinement to an open arm of the EPM and shock-probe burying tests) for up to 4 weeks after withdrawal. Depression-like behavior was assessed by the forced swim test and sucrose preference test. Altogether our results demonstrated that cocaine withdrawal induced persistent heightened levels of anxiety that last for at least 28 days but did not affect depression-like behavior. We then used Fos immunohistochemistry to map neuronal activation patterns in withdrawn rats confined to one open arm of an EPM, and a double labeling procedure using Fos immunohistochemistry and in situ hybridization of glutamic acid decarboxylase or vesicular glutamate transporter mRNAs to identify the phenotype of the activated neurons. Our data showed that the exacerbated anxiety observed in cocaine withdrawn rats exposed to an elevated open arm was accompanied by an altered reactivity of the dorsal part of the medial prefrontal cortex (anterior cingulate and dorsal prelimbic cortices), the paraventricular thalamic nucleus and the lateral and anterior areas of the hypothalamus. In the medial prefrontal cortex, we evidenced a negative correlation between Fos expression in its dorsal part and open arm-induced freezing in NaCl-treated rats but not in cocaine withdrawn rats. We also found that more than 65% of activated neurons were glutamatergic projection neurons. The present study provides new insights into the neuroanatomical regions and neuronal cell types that may underlie pathological anxiety during cocaine withdrawal.
PMCID: PMC3420872  PMID: 22916276
23.  Enhancing Spatial Memory: Anxiolytic and Antidepressant Effects of Tapinanthus dodoneifolius (DC) Danser in Mice 
We evaluated the anxiolytic and antidepressant effects of the aqueous extract of the bark of Tapinanthus dodoneifolius (TAE) (Danser) (25, 50, and 100 mg/kg), using open field, elevated plus maze, and forced swimming tests. Effect of TAE was compared to standard drugs diazepam (2 mg/kg) and imipramine (10 mg/kg). Additionally, the same doses of TAE were evaluated on rat's memory using Y-maze task. Results showed a significant (P < 0.05; 100 mg/kg) increase in the percentage of open arm entry and the time spent in the open arms in the elevated plus maze, suggesting an anxiolytic activity of the extract. In a dose-dependant manner, TAE at 25 mg/kg significantly (P < 0.05) decreased the number of lines crossed and the rearing behavior in the open field test, suggesting its possible sedative activity. In the forced swimming test, the immobility time of the animal was significantly reduced (P < 0.05) by TAE (100 mg/kg), compared to control, and this effect was quite comparable to that of imipramine. In the Y-maze paradigm, TAE at 50 mg/kg caused a significant increase in the spontaneous alternations but with a significant decrease in exploratory behavioral pattern. Taking these results together, TAE improved the spatial memory and showed anxiolytic, antidepressant, and sedative activities. The present results support the anxiolytic and antidepressant activities of TAE and, to our knowledge, for the first time, demonstrate its enhancing effect on memory.
PMCID: PMC3933263  PMID: 24649363
24.  Anxiolytic and Antidepressant-Like Effects of the Aqueous Extract of Alafia multiflora Stem Barks in Rodents 
The present study examined the anxiolytic and antidepressant effects of the aqueous extract of Alafia multiflora Stapf (AM) stem barks (150 and 300 mg/kg, 7 days administration) on rats and mice, using experimental paradigms of anxiety and depression. In the open field, the aqueous extract increased significantly the number of center square crossed and the time spent at the center of the field as well as the rearing time, while the grooming time was reduced significantly. In the elevated plus maze, the aqueous extract increased the time spent and the number of entries in the open arms. All these effects were also completely reversed by flumazenil, an antagonist of benzodiazepine receptors and pindolol a β-adrenoceptors blocker/5-HT 1A/1B receptor antagonist. The time spent in the light compartment, the latency time, and the number of the light-dark transitions increased significantly in the light/dark exploration test after the treatment with AM. The extract was able to reduce significantly the immobility time and increase swimming as well as climbing duration. Taken together, the present work evidenced anxiolytic effects of the aqueous extract of AM that might involve an action on benzodiazepine-type receptors and an antidepressant effect where noradrenergic mechanisms will probably play a role.
PMCID: PMC3485477  PMID: 23125853
25.  Pain-related anxiety-like behavior requires CRF1 receptors in the amygdala 
Molecular Pain  2007;3:13.
Corticotropin-releasing factor receptor CRF1 has been implicated in the neurobiological mechanisms of anxiety and depression. The amygdala plays an important role in affective states and disorders such as anxiety and depression. The amygdala is also emerging as a neural substrate of pain affect. However, the involvement of the amygdala in the interaction of pain and anxiety remains to be determined. This study tested the hypothesis that CRF1 receptors in the amygdala are critically involved in pain-related anxiety. Anxiety-like behavior was determined in adult male rats using the elevated plus maze (EPM) test. The open-arm preference (ratio of open arm entries to the total number of entries) was measured. Nocifensive behavior was assessed by measuring hindlimb withdrawal thresholds for noxious mechanical stimulation of the knee. Measurements were made in normal rats and in rats with arthritis induced in one knee by intraarticular injections of kaolin/carrageenan. A selective CRF1 receptor antagonist (NBI27914) or vehicle was administered systemically (i.p.) or into the central nucleus of the amygdala (CeA, by microdialysis). The arthritis group showed a decreased preference for the open arms in the EPM and decreased hindlimb withdrawal thresholds. Systemic or intraamygdalar (into the CeA) administration of NBI27914, but not vehicle, inhibited anxiety-like behavior and nocifensive pain responses, nearly reversing the arthritis pain-related changes. This study shows for the first time that CRF1 receptors in the amygdala contribute critically to pain-related anxiety-like behavior and nocifensive responses in a model of arthritic pain. The results are a direct demonstration that the clinically well-documented relationship between pain and anxiety involves the amygdala.
PMCID: PMC1891279  PMID: 17550594

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