National Journal of Physiology, Pharmacy and Pharmacology

RESEARCH ARTICLE
Comparative study of anti-nociceptive effect of venlafaxine with tramadol
by tail-flick test in animal model of mice

Sandeep Bandapati1, Karuna Sree Podila2, Venkata Rao Yadala3

Department of Medical Safety, IQVIA, Bengaluru, Karnataka, India, 2Department of Pharmacology, ESIC Medical College Hospital,
1

Hyderabad, Telangana, India, 3Department of Pharmacology, Kamineni Institute of Medical Sciences, Narketpally, Telangana, India
Correspondence to: Karuna Sree Podila, E-mail: drpkarunasri@gmail.com

Received: November 06, 2020; Accepted: February 01, 2021

ABSTRACT

Background: Pain management is ever challenging task to the treating physicians. Always there is search for new
medication for pain management. In this context, use of antidepressants in pain management had shown promising results.
Venlafaxine is a newer antidepressant drug belonging to selective norepinephrine reuptake inhibitor class, this study
was planned to evaluate the venlafaxine antinociceptive effect. Aims and Objectives: This study aims to evaluate the
venlafaxine antinociceptive activity and to compare the venlafaxine antinociceptive effect with tramadol by tail-flick test
in an animal model in albino mice. Materials and Methods: Albino mice of either sex were randomly selected with
reaction time of <6 s by tail-flick method using analgesiometer which were included in the study. Number of groups were
seven, with 6 mice in each, based on the drug received, that is, venlafaxine 15 mg/kg, 30 mg/kg, 60 mg/kg, 10 mg/kg, and
20 mg/kg of tramadol, control group (distilled water), and combination group venlafaxine (15 mg/kg) + tramadol (10 mg/
kg). Tail withdrawal latency period was calculated for each animal in group at 0, 15, 30, 60, and 90 min after administration
of drug. Results were analyzed. Results: At doses of 30 mg/kg and 60 mg/kg of venlafaxine, 20 mg/kg of tramadol, and
combination group, had shown significant (P < 0.001) increase in tail-flick latency when compared to control group
(normal saline). Venlafaxine 60 mg/kg but less potent (P = 0.86) to tramadol 20 mg/kg. Conclusion: Venlafaxine was
having anti-nociceptive effect at doses of 30 mg/kg and 60 mg/kg and was less potent than tramadol.

KEY WORDS: Venlafaxine; Tail-Flick Test; Analgesic; Chronic Pain

INTRODUCTION limb pain) or may be due to any predisposing cause (e.g.,

Pain experienced by a person varies from one to one and
difficult to define. Typically, it is a direct response associated
with tissue damage of an untoward event, such as injury,
inflammation, or cancer. Moreover, pain can persist even
after the healing of precipitated injury (e.g., phantom
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trigeminal neuralgia). Pain is defined by the taxonomy
committee of International Association for the Study of
Pain as “An unpleasant sensory and emotional experience
associated with actual or potential tissue damage or
described in terms of such damage.[1]” Several areas of the
central nervous system either directly or indirectly activated
by nociceptive inputs and implicated in central modulation
of pain. These modulatory effects are mainly mediated by
descending monoaminergic pathways that utilize serotonin,
norepinephrine, or dopamine.[2] Pain management is always
a challenge to the physician, in spite of wide availability
of different classes of drugs. Chronic pain and depression
usually coexist and evidence suggests chronic pain leads
to depression and depression leads to pain.[3] Patients with

National Journal of Physiology, Pharmacy and Pharmacology Online 2021. © 2021 Karuna Sree Podila, et al. This is an Open Access article distributed under the terms of the Creative
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Bandapati et al. Anti-nociceptive effect of venlafaxine with tramadol

chronic pain are at higher risk of depression and the presence
of pain in depressed patients had a less favorable outcome of
depression with a poor quality of life.[4]
Research for new drugs to alleviate pain is constantly there. In
this context, use of antidepressants in management of pain is
showing promising results. Tricyclic antidepressants (TCAs,
e.g., amitriptyline), serotonin norepinephrine reuptake
inhibitors (SNRIs, e.g., duloxetine), and selective serotonin
reuptake inhibitors (SSRIs, e.g., fluoxetine) are commonly
used drugs. Among the above drugs, SSRIs are less effective
in pain management when compared to the other two classes.
These drugs act by increasing serotonin levels and inhibit the
release of transmitters that carry pain sensation, thus used in
the management of pain mainly neuropathic.[5] Venlafaxine
is one of the newer antidepressant drugs belonging to the
class of SNRIs. Several preclinical studies demonstrated
the antinociceptive effect of venlafaxine, but over a variable
dose range, that is, 10 mg/kg to 60 mg/kg. As the dose may
vary depending on the species and method, the present study
planned to assess venlafaxine antinociceptive activity and its
combination with tramadol by tail-flick method, an animal
model for screening of analgesic drugs in albino mice.
Aims and Objectives
The objectives of the study were as follows:
• To evaluate the venlafaxine antinociceptive effect in
three graded doses (15 mg/kg, 30 mg/kg, and 60 mg/kg)
• To evaluate the venlafaxine antinociceptive effect in
combination with tramadol, that is, venlafaxine 15 mg/kg
+ tramadol 10 mg/kg
• To compare the antinociceptive effect of venlafaxine
with tramadol.
Drugs
Venlafaxine and tramadol used for the experiment were
purchased from Sigma-Aldrich Pharmaceuticals and Dr.
Reddy Laboratories Limited, respectively.
Equipment
Analgesiometer for the tail-flick test was used for the
evaluation of antinociceptive effect.
Study Procedure
The animals (mice) were screened for the response to pain
using analgesiometer before including them in the study. The
mice showing the response, that is, flicking the tail within
6 s, were included. The screened animals were randomly
divided into seven groups and with six animals in each group.
Grouping of the animals was mentioned in Table 1. All the
drugs were administered intraperitoneally (i.p.). For the
combination group, that is, Group 7, both drugs administered
simultaneously at different sites.
Antinociceptive effect of the drug was evaluated by tail-flick
latency period in rodent models for pain. Furthermore, the
maximum possible effect in percentage was calculated for
each group in both the models at 90 min using the below
formula.
 Post drug latency – 
 
Maximum Possible Effect  Pre drug latency  × 100
=
( MPE ) in percentage  Cut − off time – 
 
 pre drug latency 

MATERIALS AND METHODS Procedure for the Determination of Analgesic Activity

Place of Study
It was a pre-clinical randomized controlled study carried out
in the Pharmacology Department of Kamineni Institute of
Medical Sciences, after permission from Institutional Animal
Ethics Committee.
Tail-flick test
Analgesiometer is used to deliver radiant heat and this
instrument is used for testing the analgesic effect of a drug by
tail-flick response. Mice were placed into mice holder leaving
the tail exposed, that is, left outside the holder. Radiant heat
from a nichrome wire was applied to the proximal third of

Laboratory Animals Table 1: Division of groups

Healthy albino mice were procured from National Institute Name of group Drug and dose administered
of Nutrition, Hyderabad, and maintained at an ambient Group 1 Control (NS)
temperature of 25–35°C with food and water ad libitum Group 2 Tramadol 10 mg/kg
in the Central Animal House of KIMS, Narketpally, as per Group 3 Tramadol 20 mg/kg
the guidelines of Committee for the purpose of Control and Group 4 Venlafaxine 15 mg/kg
Supervision of Experiments on Animals. Healthy albino mice
Group 5 Venlafaxine 30 mg/kg
of either sex and weight of 25-35 g were included in the study
Group 6 Venlafaxine 60 mg/kg
after screening.
Group 7 Venlafaxine 15 mg/kg and tramadol 10 mg/kg

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Bandapati et al. Anti-nociceptive effect of venlafaxine with tramadol

the tail. Usually, the animal either flicks the tail or tries to
escape within a few seconds of exposure to radiant heat. The
strength of the current was kept constant at 5 ampere. Tail-
flick latency was the time duration between switching on the
analgesiometer after placing the tail over the nichrome wire
till flicking of the tail. A cutoff period of 10 s was considered
to prevent damage to the tail. The tail-flick latency period
was measured successively at 0, 15, 30, 60, and 90 min after
drug administration.
Statistical Analysis
The data were tabulated and the results were analyzed using
the SPSS software. Mean, standard deviation, analysis of
variance (ANOVA), and post hoc test (LSD) were calculated.
P < 0.05 was considered as statistically significant.
RESULTS
In this study, three graded doses of venlafaxine (15 mg/kg,
30 mg/kg, and 60 mg/kg) and combination of venlafaxine
(15 mg/kg) + sub-analgesic dose of tramadol (10 mg/kg)
were compared with analgesic dose of standard drug tramadol
(20 mg/kg) as well as with control group of normal saline
(NS) (0.1 ml).
We observed significant increase in tail-flick latency
period with venlafaxine 30 mg/kg (P < 0.001) and
60 mg/kg (P < 0.001), tramadol 20 mg/kg (P < 0.001), and
the combination group (venlafaxine 15 mg/kg and tramadol
10 mg/kg – P < 0.001) when compared to control group (NS)
(96.80 ± 2.16), and combination group (63.68 ± 3.04) when
compared to control group (4.24 ± 2.10) indicating the
antinociceptive effect in these groups.
Intergroup comparison of MPE (%) had shown that
antinociceptive effect of venlafaxine 60 mg/kg (92.5 ± 2.56)
was comparable with tramadol 20 mg/kg (96.80 ± 2.16). MPE
(%) of combination group, that is, venlafaxine 15 mg/kg with
tramadol 10 mg/kg (63.68 ± 3.04) was significantly more
than venlafaxine 15 mg/kg (3.12 ± 2.13) alone or tramadol
10 mg/kg (6.25 ± 2.28) alone indicating that venlafaxine can
potentiate antinociceptive effect of tramadol [Table 4].
DISCUSSION
Antidepressants are used effectively not only to treat
psychological and physical symptoms of depression but
also chronic pain in non-depressed patients. At present,
TCAs and SNRIs are commonly used antidepressants for
various types of chronic pain conditions. However, their
major drawback is their side effect profile. Venlafaxine,
a novel antidepressant, has similar mode of action to that
of TCAs, with favorable side effect profile.[6-8] Proper
assessment of analgesic potency and efficacy of venlafaxine
may replace TCAs in the management of chronic pain in
future. The results of this study have shown increase in
tail-flick latency at 30 min, 60 min, and 90 min intervals
12
Control (NS)
10
Tail flick latency (in seconds)

Tramadol (10 mg/kg)

at 30, 60, and 90 minutes. However, significant difference
8
was not observed in tail-flick latency period with venlafaxine Tramadol (20 mg/kg)
15 mg/kg and tramadol 10 mg/kg when compared to control 6 Venlafaxine (15 mg/kg)
group [Table 2 and Figure 1]. It indicates that venlafaxine
at doses of 30 mg/kg and 60 mg/kg, tramadol at 20 mg/kg, 4 Venlafaxine (30 mg/kg)

and combination of venlafaxine 15 mg/kg with tramadol Venlafaxine (60 mg/kg)

10 mg/kg were having antinociceptive effect. 2
Venlafaxine 15 mg/kg +
0 Tramadol 10 mg/kg
The percentage of maximal possible effect (MPE) in tail- 0 min 15 min 30 min 60 min 90min

flick latency at 90 min was calculated [Table 3]. We observed Duration of time(in minutes)from
administration of drug
significant difference in venlafaxine 30 mg/kg (61.80 ± 3.44),
venlafaxine 60 mg/kg (92.5 ± 2.56), tramadol 20 mg/kg Figure 1: Tail flick latencies expressed in terms of Mean ± SEM

Table 2: Tail-flick latencies expressed in terms of mean ± standard error of the mean (SEM) at regular intervals
Groups 0 min 15 min 30 min 60 min 90 min
Control (NS) 2.17±0.10 2.17±0.10 2.25±0.11 2.25±0.11 2.42±0.15
Tramadol (10 mg/kg) 2.17±0.10 2.25±0.11 2.33±0.17 2.50±0.13 2.67±0.10
Tramadol (20 mg/kg) 2.17±0.10 2.58±0.15 6.58±0.15* 8.59±0.15* 9.75±0.17*
Venlafaxine (15 mg/kg) 2.25±0.11 2.25±0.15 2.33±0.17 2.42±0.15 2.42±0.15
Venlafaxine (30 mg/kg) 2.17±0.10 2.25±0.11 4.92±0.24* 6.00±0.22* 7.00±0.29*
Venlafaxine (60 mg/kg) 2.25±0.11 2.33±0.10 6.50±0.13* 8.00±0.18* 9.42±0.20*
Venlafaxine 15 mg/kg + tramadol 10 mg/kg 2.17±0.10 2.25±0.11 5.17±0.28* 6.33±0.17* 7.17±0.21*
*P<0.001. NS: Normal saline

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with venlafaxine 30 mg/kg and 60 mg/kg dose indicating
the analgesic effect of venlafaxine. Combination treatment
of low-dose venlafaxine 15 mg/kg with tramadol 10 mg/kg
increased tail-flick latency period at 30, 60, and 90 min in
comparison to control (NS) treatment group, venlafaxine 15
mg/kg, and tramadol 10 mg/kg alone. Calculated MPE of
tail-flick latency in percentage (%) at 90 min suggested that
venlafaxine (30 mg/kg and 60 mg/kg), tramadol 20 mg/kg,
and combination group produced significant antinociceptive
effect in the tail-flick test. Further, intergroup comparison
of MPE (%) has shown that antinociceptive effect of
venlafaxine 60 mg/kg was comparable with tramadol
20 mg/kg, indicating that venlafaxine is less potent than
tramadol. MPE (%) in combination group (venlafaxine
Table 3: Comparison of mean ± S.E. of MPE (%) of tail-
flick latency
Groups Mean±SE
Tramadol (10 mg/kg) 6.25±2.28
Tramadol (20 mg/kg) 96.80±2.16
Venlafaxine (15 mg/kg) 3.12±2.13
Venlafaxine (30 mg/kg) 61.80±3.44
Venlafaxine (60 mg/kg) 92.50±2.56
Venlafaxine 15 mg/kg + tramadol 10 mg/kg 63.68±3.04
15 mg/kg with tramadol 10 mg/kg) was significantly
higher when compared to venlafaxine (15 mg/kg alone)
or tramadol (10 mg/kg alone), indicating that venlafaxine
can potentiate antinociceptive effect of tramadol. Thus by
combining low doses of both drugs, analgesic effect can be
achieved as well as adverse effects occurring at high doses
may be avoided.
The study results were in par with studies done by Jha et al.
in tail-flick and writhing test analgesic models in albino mice
(2006)[9] and Sikka et al. in tail-flick model in mice (2011)[10]
who also reported the anti-nociceptive effect of venlafaxine
but at slightly different doses [Table 5]. Jha et al. (2006),[9]
Wrzosek et al. (2003),[11] and Uyar et al. (2003)[12] also
reported potentiation of antinociceptive effect of tramadol
with venlafaxine in animal models for analgesic effect.
Strengths
It was a well-planned study and study design with doses of
venlafaxine different from the other studies. Venlafaxine was
having antinociceptive activity at doses of 30 mg/kg and
60 mg/kg and in combination (venlafaxine 15 mg/kg with
Tramadol 10 mg/kg) and the effect observed was at slightly
different dose compared to other studies.

Table 4: Intergroup comparison of MPE in % of tail-flick latency by multiple comparison (LSD) test
Comparison between different groups Mean difference P-value
CTRL versus TRA 10 −2.01 0.58 (ns)
CTRL versus TRA 20 −92.56 0.0001***
CTRL versus VEN 15 1.11 0.76 (ns)
CTRL versus VEN 30 −57.57 0.0001***
CTRL versus VEN 60 −88.26 0.0001***
CTRL versus VEN 15+TRA10 −62.78 0.0001***
TRA 10 versus TRA 20 −90.55 0.0001***
TRA 10 versus VEN 15 3.12 0.40(ns)
TRA 10 versus VEN 30 −55.55 0.0001***
TRA 10 versus VEN 60 −86.25 0.0001***
TRA 10 versus VEN 15+TRA10 −57.43 0.0001***
TRA 20 versus VEN 15 93.67 0.0001***
TRA 20 versus VEN 30 34.99 0.0001***
TRA 20 versus VEN 60 4.30 0.25(ns)
TRA 20 versus VEN 15 + TRA 10 33.12 0.0001***
VEN 15 versus VEN 30 −58.68 0.0001***
VEN 15 versus VEN 60 −89.37 0.0001***
VEN 15 versus VEN 15 + TRA 10 −60.55 0.0001***
VEN 30 versus VEN 60 −30.69 0.0001***
VEN 30 versus VEN 15 + TRA 10 -1.87 0.61(ns)
VEN 60 versus VEN 15 + TRA 10 28.82000 0.0001***
Significance of P < 0.001***, < 0.01**, < 0.05*, > 0.05 = not significant (ns)
CTRL – Control, TRA 10 – Tramadol 10 mg/kg, TRA 20 – Tramadol 20 mg/kg, VEN 15 – Venlafaxine 15 mg/kg, VEN 30 – Venlafaxine 30 mg/kg, VEN 60 –
Venlafaxine 60 mg/kg, VEN 15 + TRA 10 – Venlafaxine 15 mg/kg + tramadol 10 mg/kg

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Bandapati et al.
Table 5: Comparison of tail-flick latency period (mean±SD) of the previous studies with venlafaxine pre-treatment at 0
min and 60 min
Groups Jha et al. (mean±SD)
0 min 60 min
Venlafaxine 10 mg/kg 3.73±0.33 5.6±0.66
Venlafaxine 22.5 mg/kg 3.41±0.06 5.86±0.05
Venlafaxine 15 mg/kg
Venlafaxine 30 mg/kg
Venlafaxine 50 mg/kg
Venlafaxine 60 mg/kg
Venlafaxine 15 mg/kg + tramadol 10 mg/kg
Limitations
The antinociceptive evaluation in neuropathic animal
models was not assessed. The safety of drugs as well as the
mechanism of action was not studied.
It was presumed that venlafaxine antinociceptive effect might
be due to inhibition of serotonin-norepinephrine reuptake,
leading to NE/5-HT accumulation. Further, descending
5-HT and NE pathways are known to have analgesic action
by inhibiting pain causing afferent impulses. In addition, it
has opioid like activity also. The mechanism of the action
of venlafaxine analgesic effect was not studied in our study.
CONCLUSION
The present study suggests that venlafaxine, a SNRI, has
antinociceptive activity, which was evaluated by tail-flick
test in albino mice, an analgesic animal model. Venlafaxine
is less potent antinociceptive agent than tramadol and
potentiates the antinociceptive activity of standard drug
tramadol when both are given in substandard doses. Further
experimental and human studies are required to confirm the
antinociceptive activity of venlafaxine.
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How to cite this article: Bandapati S, Podila KS, Yadala VR.
Comparative study of anti-nociceptive effect of venlafaxine
with tramadol by tail-flick test in animal model of mice. Natl J
Physiol Pharm Pharmacol 2021;11(06):633-637.
Source of Support: Nil, Conflicts of Interest: None declared.
2021 | Vol 11 | Issue 06