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Study on anti-anxiety effect of Willughbeia cochinchinesis in scopolamine-administrered mice using open field test

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The aim of this study is: To investigate effects of a new natural plant, Wullughbeia cochinchinensis (WC) to meliorate deficit in anxiety-like behaviors in mice by using the animal model.

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STUDY ON ANTI-ANXIETY EFFECT OF WILLUGHBEIA COCHINCHINENSIS IN SCOPOLAMINC-ADMINISTERED MICE

BY USING OPEN FIELD TEST

Nguyen Tat Dinh*; Cao Tien Duc*; Le Van Quan**

SUMMARY

Objective: To investigate effects of Wullughbeia cochinchinensis to ameliorate disorders in anxiety-like behaviors of mice Subjects and methods: In the present study, 50 Swiss mice were induced anxiety-like behaviors by scopolamine Behaviors of mice treated with and without Wullughbeia cochinchinensis at doses of 100 mg/kg, 150 mg/kg and 200 mg/Kg and/or scopolamine were tested using the open field test Results: Wullughbeia cochinchinensis at dose of 150 mg/kg and 200 mg/kg induced a significant increase in time spent, number of entries and travel distances in the center area of open field in animals with scopolamine-induced anxiety-like behaviors Conclusion: Wullughbeia cochinchinensis 150 mg/kg and 200 mg/kg might ameliorate deficits in anxiety like symptoms in experimental animals

* Keywords: Anxiety, Mice; Willughbeia cochinchinensis

INTRODUCTION

Anxiety is one of the common symptoms

in mental disorders such as depression,

phobia, etc In a recent study, it has

suggested that 33.7% of the population

are affected by an anxiety disorder during

their lifetime [1] Furthermore, ratio of

females with anxiety disorder is higher

than this of male with this symptom [2] In

treatment, benzodiazepines have been

suggested to be effective to anxiety

disorders However, these drugs might

induce drug dependence when they are

used a long time [3] Thus, it is necessary

to investigate new natural plants or drugs

for treating anxiety disorders

It has suggested that anxiety-like

disorders might relate to activities of

achetylcholine Thus, scopolamine, an antagonist to acetylcholinergic receptors, might be used to induce anxiety-like behaviors in experimental animals [4] In the present study, we used this animal model to investigate effects of a new natural plant, Wullughbeia cochinchinensis (WC) to meliorate deficit in anxiety-like behaviors in mice

SUBJECTS AND METHODS

1 Subjects

50 Swiss mice (150 - 250 g body weight) were used in the present study Animals were housed in individual cages, maintained in controlled temperature and

12 h light/dark cycles with free access to water and food

*

**

Corresponding author: Le Van Quan (@gmail.com)

Date accepted: 30/05/2018

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Animals were separated randomly into

5 experimental groups, 10 mice for each

group: group 1 (control group): mice were

ip and p.o treated saline; group 2

(scopolamine group): mice were i.p

treated scopolamine 1.5 mg/Kg and p.o

treated saline at 0.1 ml/10 g; group 3,

group 4 and group 5 (WC groups): mice

were i.p injected scopolamine 1.5 mg/Kg

and p.o WC 100 mg/Kg, 150 mg/Kg and

200 mg/Kg, respectively WC and saline

were orally administered at 60 minutes

and scopolamine and saline were i.p

injected at 30 minutes before the

behavioral task

The present study was conducted at

Department of Physiology, Vietnam

Military Medical University All procedures

were performed in accordance with the

Animal Center Guidelines for the Care

and Use of Laboratory Animals at the

Vietnam Military Medical University

2 Materials

- WC was isolated by Department of

Pharmacy, Hochiminh City University of

Medicine and Pharmacy and was

supplied in power form WC power was

dissolved in saline using a magnetic

stirrer

- Open field box was a square box (40

x 40 x 60 cm), covered with polypropylene

sheets inside the wooden box In the

present study, the open field was

separated into two areas: a center area

and a periphery zone (figure 1)

Figure 1: Open field box

3 Methods

* Open field test:

60 minutes after drug treatments, mice were placed in the center of an open field box Animals were allowed to free explore inside open field box for 5 minutes Behaviors of animals were recorded using

a digital video system Data was analyzed offline by ANY-maze software (Stoelting Co., Wood Dale, IL, USA)

* Research indicators:

To investigate anxiety-like behaviors in mice, we concentrated to analyze activities

of animals in the center area of the open field apparatus These were:

- Time spent in the central zone (s)

- Numbers of entries into the central area

- Travel distances in the central area (m)

* Data analysis:

Time spent in the central area; numbers of entries in the central zone and travel distances in the center area were analyzed by one-way analysis of variance (ANOVA) followed by the Tukey’s post-hoc test for multiple comparisons, using SPSS 19.0 Results were considered to

be statistically significant at p < 0.05 All results were expressed as mean ± SEM

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RESULTS

1 Time spent in the central area of the open field

Figure 2: Time spent in the center area

Time spent in the central area is used to assess exploring abilities as well as anxiety-like behaviors of mice In the fig 2, time spent in the central area of animals treated by scopolamine was significant shorter than this of animals treated by saline (p

< 0.05) After animals treated by WC, there were gradual increases in time spent in the central area However, significant differences were found only in animals treated by

WC at doses of 150 mg/kg and 200 mg/kg (p < 0.05)

2 Numbers of entries to the central area

Figure 3: Entries to the central area

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Fig 3 indicated results of entries of

mice into the central area Results

showed that number of entries into the

central area of mice treated by

scopolamine was significant lower than

this of mice treated by saline (control

group) (p < 0.01) Furthermore, there

were gradual increases in entries of mice

into central areas following treatments of

WC at doses of 100 mg/kg; 150 mg/kg

and 200 mg/kg However, there were

significant differences in numbers of

entries into the central area of mice

treated by WC at doses of 150 mg/kg and

200 mg/Kg (p < 0.05)

3 Travel distances in the central area,

Fig 3 showed travel distances in the central areas of mice in experimental groups Results showed that mean travel distance in the central area of scopolamine-treated mice was significant shorter than this of saline-treated mice (p < 0.01) After treatments of WC 60

minutes, travel distances in the central area of mice increased gradually

However, travel distances of mice in the central areas increased significantly

in groups WC 150 and WC 200 only (p < 0.01)

Figure 4: Travel distances in the central area

DISCUSION

Open field is one of the common

behavioral tests which are employed to

investigate anxiety-like disorders in rodent

animals In the open field test, the time

animals spent and number of entries to

the center of arena is often used to

measure anxiety-related behaviors in mice In general, due to life characteristics

of mice, they avoid the center of open field arena Increases in time spent and numbers of entries into the central areas

of the open fields might indicate a decrease in anxiety behaviors [5]

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In the present study, we found evidence

of improving anxiety disorder effect of WC

These were: treatments of WC at doses

of 150 mg/kg and 200 mg/kg induced

increases in time spent, numbers of

entries and travel distances in the center

of open field arena These results are

consistent with previous studies which

have concentrated to investigate anxiety

disorders in mice Anchan et al showed

that GPR30 activation decreases anxiety

in the open field which was expressed by

a greater distance and higher number of

entries into the central area in the open

field [6] Similarly, by the some way, they

have demonstrated that γ-aminobutyric

acid transporter-1 also is involved in

decrease of anxiety like behaviors in mice

[7] From above results, our study showed

a direct evidence for effects of WC to

decreasing anxiety like behaviors in mice

CONCLUSION

In the present study, we investigated

anti-anxiety effect of WC in mice We

found that: After treatments of WC at

doses of 150 mg/kg and 200 mg/kg

increase in time spent in the center of

arena, greater travel distances and higher

numbers of entries into the central of

open field area These results provided

new evidences for using new natural

plants in treatments for anxiety disorders

in humans

REFERENCES

1 Bandelow B Epidemiology of anxiety

disorders in the 21st century Dialogues Clin Neurosci 2015, 17 (3), pp.327-335

2 McLean C.P, Asnaani A, Litz B.T, Hofmann S.G Gender differences in anxiety

disorders: Prevalence, course of illness, comorbidity and burden of illness J Psychiatr Res 2011, 45 (8), pp.1027-1035

3 Starcevic V Benzodiazepines for

anxiety disorders: maximizing the benefits and minimizing the risks Advances in Psychiatric Treatment 2012, 18 (4), pp.250-258

4 Rodgers R.J, Cao B.J, Dalvi A, Holmes

A Animal models of anxiety: an ethological

perspective Braz J Med Biol Res 1997, 30 (3), pp.289-304

5 Walsh R.N, Cummins R.A The

open-field test: a critical review Psychological Bulletin 1976, 83, pp.482-504

6 Anchan D, Clark S, Pollard K, Vasudevan N GPR30 activation decreases

anxiety in the open field test but not in the elevated plus maze test in female mice Brain Behav 2014, 4 (1), pp.51-59

7 Gong X, Shao Y, Li B, Chen L, Wang C, Chen Y γ-aminobutyric acid transporter-1 is involved in anxiety-like behaviors and cognitive function in knockout mice Exp Ther Med 2015, 10 (2), pp.653-658

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