EVALUATION OF ANTIDEPRESSANT ACTIVITY OF TRAMADOL AND TRAMADOL PLUS IMIPRAMINE USING RESERPINE...

Inter. J. of Phytotherapy / Vol 3 / Issue 1 / 2013 / 18-23.

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e - ISSN - 2249-7722

Print ISSN - 2249-7730

International Journal of Phytotherapy

www.phytotherapyjournal.com

EVALUATION OF ANTIDEPRESSANT ACTIVITY OF TRAMADOL

AND TRAMADOL PLUS IMIPRAMINE USING RESERPINE

INDUCED HYPOTHERMIA MODEL ON EXPERIMENTAL

ANIMALS

Mohd Riyaz1, Vinit Raj

1, Ashish Kumar

2, Satyajeet Singh

2

1,2Roorkee College of Pharmacy (RCP Universe) 09 milestone Roorkee-Dehradun Highway, Vill. Kishanpur, P.B.NO.104,

Roorkee, Distt. Haridwar Uttarakhand - 247667, India.

INTRODUCTION

The cause of depression was linked to

decreased brain levels of the neurotransmitters NE, 5-

HT, and DA, although the actual cause remains unknown

[1, 2]. Although the reuptake blockade of monoamines

(e.g., NE and 5-HT) occurs immediately upon

administration of an antidepressant, the clinical

antidepressant effects generally are not observed until

after 4 weeks of dosing It is apparent that no single

neurotransmitter theory of depressions adequate.[3, 5].

The 5-HT or NE link hypothesis maintains that both the

serotonergic and noradrenergic systems need to be

functional for an antidepressant effect to be exerted [6].

The 5-HTor NE link hypothesis is also consistent with

the rationale of the postsynaptic alteration theory of

depression, which emphasizes the importance of alpha

adrenergic receptor downregulation for achieving an

antidepressant effect [7]. Again, it has been proposed

that both NE and 5-HT are necessary for homologous

desensitization of central a-adrenergic receptors by

antidepressants. Tramadol is a synthetic centrally acting

opioid analgesic used mainly for the treatment of

moderate to severe pain. It is a weak opioid receptor

agonist and also produces analgesia by inhibiting uptake

of norepinephrine and serotonin [8].

MATERIALS AND METRHODS

All the drugs used in present study were of

pharmaceutical grade. Tramadol was gifted by Pro-

laboratories Pvt, Ltd, Roorkee, India, used as a test drug. All solvents were of analytical grade and procured from

Corresponding Author:- Mohd Riyaz Email: [email protected],

ABSTRACT

The present study was carried out to evaluated antidepressant activity of Tramadol and Tramadol plus

Imipramine in experimental animals by using Reserpine Induced Hypothermia Test. Standard drug used in tests was

Imipramine (10 mg/kg p.o.). It is a tricyclic antidepressant and it is a monoamine reuptake inhibitor. It inhibits the

reuptake of noradrenalin and serotonin (5-HT) by monoaminergic nerve terminals, and thus it facilitates transmission.

So, the possible mechanism of Tramadol and combination of Tramadol and Imipramine to decrease the immobility

time may be due to the inhibition of monoamine reuptake. These models suggest that Tramadol and combination of

Tramadol and Imipramine increases availability of biogenic amines Hence, Tramadol may produces antidepressant

due to inhibition of reuptake of noradrenalin and 5-HT (serotonin).

Key words: Serotonin, Dopamine transport, Biogenic amines, Reserpine Induced Hypothermia Test, Imipramine, and

Tramadol.


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Division of Pharmaceutical Sciences, Shri Guru Ram Rai

Institute of Technology and Science, Patel Nagar

Dehradun. Reserpine was purchase from Rajesh

Chemicals Mumbai, India and used for induction of

hypothermia. Imipramine was gifted from Unicure

Pharmaceuticals Pvt Ltd, Roorkee India.

Animals

Swiss albino mice weighing between 2025g. All the animals were acclimatized at least under standard

husbandry conditions, i.e. room temperature of 24 1 C;

relative humidity 45 55% and a 12 : 12 h light/dark cycle. The animals were free access to standard pellets of

rat with water supplied under strict hygienic conditions.

Each experimental group was separate set of animals and

care was taken to ensure that animals used for one

response was not employed anywhere. Animals were

habituated to laboratory conditions for 48 hours prior to

experimental protocol to minimize if any non- specific

stress [9, 13].

Experimental protocol

Experimental designs for Reserpine Induced

Hypothermia Test.

Animals divided in to nine groups and each group

containing six animals (n=6).

Group 1- (Negative control) Give Saline (1ml/100gm

body wt.)

Group 2-(Positive control) Give Reserpine (2mg/kg

body wt. s.c.)

Group 3-(Standard control) Give Reserpine (2 mg/kg

body wt. s.c.) and after 18 hr give Imipramine (10 mg/kg

body wt. p.o.)

Group 4- Give Reserpine (2 mg/kg body wt. s.c.) and

after 18 hr give Tramadol (10 mg/kg body wt.p.o.)

Group 5- Give Reserpine (2 mg/kg body wt. s.c.) and

after 18 hr give Tramadol (20 mg/kg body wt.p.o.)

Group 6- Give Reserpine (2 mg/kg body wt.s.c.) and

after 18 hr give Tramadol (40 mg/kg body wt. p.o.)


after 18 hr give Imipramine (5 mg/kg body wt. p.o.) +

Tramadol (5mg/kg body wt.p.o.)



Tramadol (10mg/kg body wt.p.o.)



Tramadol (20mg/kg body wt. p.o.)

METHODS

Reserpine induced hypothermia

Depletion of biogenic amines (noradrenalin, 5-

hydroxytryptamine, and dopamine) in the brain induces

not only catalepsy and apoptosis but also hypothermia in

rodents. The decrease of body temperature induced by

Reserpine is antagonized by antidepressants, MAO-

inhibitors and central stimulants. The subcutaneous

administration of 2 mg/kg Reserpine leads to a decrease

of core temperature in mice to 2023 C after 18 h. The fall in temperature can be antagonized by antidepressants.

Procedure

Groups of 6 Swiss albino mice (20-25 g body

weight) were used. On the day before testing, they were

dosed with 2 mg/kg Reserpine s.c. They were housed in a

climate controlled animal colony and have free access to

food and water. Eighteen hours after Reserpine

administration, the animals were placed into individual

cages. The initial rectal temperature was determined by

insertion of an electronic thermometer to a constant depth

of 2 cm. Following administration of the test compound

the rectal temperature were measured again at 60 min

intervals for 7 hrs [14].

Evaluation

Rectal temperature was recorded every hour. The

difference in temperature from vehicle controls were

calculated for each time and the maximal difference were

scored. The differences will be then statistically

compared.

Statistical analysis

The values were expressed as mean SEM. The

results were subjected to statistical analysis by using one-

way ANOVA. Followed by Bonferroni Test, P


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Table 1. Effect of Tramadol (p.o.) on Body Temperature in the Reserpine Induce Hypothermia Test using Swiss albino mice.

S.

No. Treatment Basal Temp.

Basal temp.

(oC) after 18

hrs.

of Reserpine

Treatment

Temperature after treatment (oC)

1 hrs 2 hrs 3 hrs 4 hrs 5 hrs 6 hrs 7 hrs

1 Negative

Control 37.760.16 37.580.16

37.46

0.10### @@

37.44

0.09@@@

37.4

20.

08 @@@

37.52

0.1

6 @@@

37.56

0.04@@@

37.36

0.09@@@

37.76

0.07@@@

2 Positive

Control 37.70.11 35.530.15

35.54

0.12 ***

##

35.31

0.06*

**##

35.4

10.

10***

###

35.35

0.1

6***

##

#

35.76

0.12*

**###

36.12

0.19*

**###

36.34

0.17*

**###

3

Standard

(IMP

10mg/kg)

37.660.20 35.610.20

36.23

0.12 ***

36.66

0.09*

**@@@

37.0

7

0.07 @@@

37.38

0.0

6 @@@

37.68

006@@@

37.67

0.14@@@

37.71

0.15

4 TMD

(10mg/kg) 37.680.16 35.620.18

35.89

0.08 ***

36.37

0.18*

**@@@

36.7

0.17*

*

@@@

37.06

0.1

6 @@@

37.48

0.18@@@

37.83

0.16@@@

37.97

0.15@@@

5 TMD

(20mg/kg)

37.780.13

35.560.13

35.99

0.11 ***

36.49

0.11*

**@@@

36.7

3

0.11*

*

@@@

37.21

0.12 @@@

37.61

0.09@@@

37.80

0.08@@@

38.08

0.07@@@

6 TMD

(40mg/kg)

37.840.11

35.330.12

36.06

0.12 ***

36.79

0.12*

@@@

37.4

1

0.09 @@@

37.86

0.09 @@@

38.07

0.12@@@

37.99

0.16@@@

37.99

0.14@@@

IMP is Imipramine as a standard drug and TMD is Tramadol as a test drug. When compared with Negative Control group (p<

0.05 = *, p


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3

Standard

(IMP

10mg/kg)

37.660.2

0 35.610.20

36.23

0.12 ***

36.66

0.09*

**@@@

37.07

0.07 @@@

37.38

0.06 @@@

37.68

006@

@@

37.67

0.14@@@

37.71

0.15

4

IMP+TMD

(5mg+5mg/k

g)

37.810.1

3

35.480.13

35.89

0.12 ***

36.31

0.13*

@@@

36.70

0.15

*

*

@@@

37.05

0.13 @@@

37.43

0.14@@@

37.68

0.12@@@

37.96

0.12@@@

5

IMP+TMD

(5mg+10mg/

kg)

37.820.1

1

35.340.11

35.92

0.03 ***

36.46

0.12*

**@@@

37

0.11 @@@

37.49

0.12 @@@

37.81

0.15@@@

37.91

0.15@@@

38.06

0.14@@@

6

IMP+TMD

(5mg+20mg/

kg)

37.780.1

2

35.350.13

36.15

0.04 *** @@

36.91

0.10*

**@@@

37.43

0.13 @@@

37.97

0.15

# @@@

38.08

0.15@@@

38.12

0.13*

*@@@

38.21

0.14@@@

IMP is Imipramine as a standard drug and TMD is Tramadol as a test drug. When compared with Negative Control group (p<

0.05 = *, p


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Figure 2. Effect of combination of Tramadol and Imipramine (p.o.) on Body Temperature in the Reserpine Induce

Hypothermia Test using Swiss albino mice

The effect of combination of Imipramine and Tramadol (5+5, 5+10, 5+20 mg/kg p.o.) and Imipramine 10mg/kg p.o. in the

five minute intervals on Reserpine induced Hypothermia Test. The results were analyzed for statistical significance using

one-way ANOVA followed by Bonferroni test. p


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EVALUATION OF ANTIDEPRESSANT ACTIVITY OF TRAMADOL AND TRAMADOL PLUS IMIPRAMINE USING RESERPINE...

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