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International Journal of Pharmaceutical  Biological Archives 2020; 11(2):91-98
ISSN 2582 – 6050
RESEARCH ARTICLE
Effects of Noise Stress on Body Weight and Adrenal Gland Weight of Male
Wistar Rats
Ravinder Kr Mehra1,2
*, Mahesh Prasad3
, Dinesh Kr Sharma4
, Ravinder Kr Mehra2
1
Department of Pharmacology, School of Pharmacy, Bharat Institute of Technology, Meerut, Uttar Pradesh, India,
2
Department of Pharmaceutical Sciences, IFTM University, Moradabad, Uttar Pradesh, India, 3
Department of
Pharmacology, Kamla Nehru Institute of Management and Technology, Sultanpur, Uttar Pradesh, India, 4
Department of
Pharmaceutical Sciences, Himalayan Institute of Pharmacy and Research, Uttarakhand, Uttar Pradesh, India.
Received: 15 March 2020; Revised: 28 April 2020; Accepted: 10 May 2020
ABSTRACT
The present study was designed to investigate the effects of varying intensities of noise stress onto the
body and adrenal gland weight (absolute) of 17 weeks old (291–296) gram (g) Wistar rat. Animals were
exposed to varying intensity of noise, i.e., 60 dB (Decibel), 80 dB, 100 dB, 120 dB, and 140 dB for
1 h (h)/day for 30 days. The dose of different noise intensities was delivered through a special fabricated
noise chamber which had provided all daily needs of animal-like oxygen, food, and water. Food and water
consumption behavior was recorded on 1st
, 14th
, and 29th
days and body weight change recorded on 1st
,
15th
, and 30th
days after the exposure of noise stress. Absolute adrenal glands weight was recorded on the
30th
day. The results of the present study showed that the food, water consumption behavior, and body
weight (g) of the experimental group EG1
(60 dB), EG2
(80 dB), EG3
(100 dB),
EG4
(120 dB), and EG5
(140 dB) were found to be significantly (P  0.001) decreased during 1st
–14th
days and during 15th
–30th
days a recovery which were observed in food, water consumption, and in body weight of the EG1
(60 dB),
EG2
(80 dB), and EG3
(100 dB). A percentage (%) decrease was found, i.e., 0.76, 0.37, 3.87, 25.89, and
30.06% in body weight of EG1
(60 dB), EG2
(80 dB), EG3
(100 dB),
EG4
(120 dB), and EG5
(140 dB)
experimental groups, respectively.
Keywords: Body weight, decibel, noise stress
INTRODUCTION
In the current scenario, human and animal health
affected by uncountable stimuli and these stimuli
may responsible for the progression of several
health issues. At present, we are surrounded by
an environment that is filled with uncountable
stressors.
Stress can be defined as; it may a kind of physical,
chemical, and emotional stimuli to which an
individualfailstomakeasuitableadaptation.[1]
Stress
is a condition that aggravated by psychological,
*Corresponding Author:
Ravinder Kr Mehra,
E-mail: rxravimehra@gmail.com
physiological, or environmental stressors and
could insult the homeostasis. It provokes the
hypothalamic-pituitary-adrenal (HPA) axis and
the sympathetic nervous system causing in a
physiological change.[2]
Our surrounding is layered
with so many types of stress, from all others, noise
stress intentionally (any loud concert) and non-
intentionally (occupational) is regarded as the
most common stimuli. Noise is regarded to every
surplus sound generated by natural phenomena
(e.g., wind, volcanic eruption, and oceans) or
human-based sources (e.g., automobiles, machines,
and explosions).[3,4]
and environmental stress
that is capable of causing hearing impairment,
behavioral, mental, and physiological alterations.[5]
Many epidemiological studies established a clear
Mehra, et al.: Effects of noise stress on male wistar rats
IJPBA/Apr-Jun-2020/Vol 11/Issue 2 92
connection between noise and vascular diseases
including hypertension[6,7]
and coronary heart
disease.[8,9]
Even, constant exposure to high levels
of sound, for more than 5 years, may eventually
lead noise-induced permanent threshold shift
and can lead to elevated stress levels as well as
stimulate violent behavior;[10]
likewise, explosive
noise may produce a massive adverse effect on the
human body and mind.[11]
Continuous noise stress
can severely insult the homeostasis and this may
initiate the events of cellular injury if the injury
may stay continue due to persistent stress could
open the way for several disease pathogenesis.
It has been known that peptic ulcers occurred much
morefrequentlyinwartimethaninpeacetime.[12]
The
study revealed that due to high levels of noise stress
variety of health issues including cardiovascular
diseases,[13]
mental illness,[14]
sleep disorder,[15]
immune function,[16]
hormonal levels,[17]
respiratory
systems,[18,19]
and hypertension occurred.
Furthermore, gastrointestinal motility disorder,
gastritis, and peptic ulcers have been noticed in
populations[2]
because noise acts as a non-specific
biological stressor and causes dysfunction of body
organs and systems[20]
and widespread disturbances
at several levels in human organs and apparatus
due to chemical and physiological modification of
endocrine.[21]
Noise stress of any kind is known to be a stress
factor which alters some physiological and
behavioral changes in human as well as animals.
The present work is designed to study the effect of
noise stress on animal body weight and the adrenal
gland weight.
MATERIALS AND METHODS
Experimental animals
17 weeks old male Wistar rats weighing around
291–296 g were used for the present study. The
research project was initiated after obtaining
clearance from the Institutional Animal Ethical
Committee of Kamla Nehru Institute of
Management and Technology, Sultanpur, Uttar
Pradesh, India. All procedures for housing and
caring of animals were followed according to the
guidelines of the ethical committee. Animals were
acclimatized in normal animal house conditions for
a week before starting the experimental protocol.
Wistar rats were housed in an open wire cage in
a temperature-controlled room (22–24°C) and
12 h light-dark cycle with free access to standard
laboratory rat chow and tap water. To minimize all
undesired stressors, such as handling, and habitat,
animals were not exposed to noise for at least
1 week after delivery to the experimental room.
Noise treatment
Arectangular special noise chamber was fabricated
to create a noise environment inside by fixing two
adjacent speakers with amplifier and a noise meter
was set to record and a controller knob was fixed
to ensure the delivery of desired sound intensities
in dB. A transparent acrylic front was provided
to the noise chamber for viewing the activity of
animals during experimentation. The chamber
had all provisions for water, feed, and aeration.
Before starting, the procedure animals were left
inside the noise chamber without sound to get
accustomed to the interior of the chamber during
the acclimatization period.
Study design
Male Wistar rats of 17 weeks old (291–296 g)
were included in this study. The animals were
divided into five subgroups (n=6) as EG1
, EG2,
EG3,
EG4
, and EG5
based on noise stress exposure
of for 1 h/day for 30 days, respectively. Every
five groups were considered a statistical unit
in this study in case of investigating effects of
noise onto body weight. The time of noise stress
exposure (9:00 AM–10: AM) was kept constant
for each exposed group throughout the end of the
experiment. Noise stress of 60, 80, 100, 120, and
140 dB intensities were delivered for 1 h/day for
30 day through a specially designed noise chamber
to EG1
, EG2,
EG3,
EG4
, and EG5
experimental
groups. To avoid age variation between all groups,
an unbiased experimental protocol was decided
to conduct and total study was conduct in total
150 days (30 days for each group) so, initially
EG1
group was exposed to 60 dB noise stress for
Mehra, et al.: Effects of noise stress on male wistar rats
IJPBA/Apr-Jun-2020/Vol 11/Issue 2 93
1 h/day for 30 days and then the reaming EG2,
EG3,
EG4
, and EG5
groups were separately exposed to
noise stress of 80, 100, 120, and 140 dB in 120
days, respectively.
A normal control group was also constructed to
evaluate the absolute weight of adrenal glands (g)
and for comparing food and water consumption of
EG1
, EG2,
EG3,
EG4
, and EG5
experimental groups,
respectively, whereas in case of evaluating the
effect of noise stress on the body weight in EG1
,
EG2,
EG3,
EG4, and EG5 experimental groups,
each group was considered a statistical unit and
compared with EG1
(60 dB) group on 1st
, 15th
, and
30th
days.
Water and food intake observation
Consumption of the food and water intake was
recorded of the experimental animal group after
delivery to the experimental room without access
to noise stress and fed a homogeneous chow diet.
A per-day analysis was performed to calculate
food and water consumption in 1 week. During
this acclimatization, per day food and water
consumption was recorded. After the 1 week of
acclimatization, animals have attained an age of 17
weeks and their average weight was found to be
291–296 g. Exposure to noise stress was started at
the age of 17 weeks. On the 0th
day, the initial body
weight of each animal was recorded and exposure
of noise stress started. A per-day analysis of food
and water consumption behavior was recorded to
evaluate the effect of noise stress onto the same
but final interpretation was done with results of 1st
,
14th
, and 29th
days consumption.
Measurement of body weight and adrenals
weight
Weight measurement was done on 1st
, 15th
, and 30th
days with the help of an electronic weighing scale
of the group under experimentation. Exposure of
noise was started on the 1st
day after recording
the initial body weight. On the 29th
day, all
animals were deprived of food and had to access
only water in their habitat. On the 30th
day, after
recording the final weight, animals were sacrificed
as per the norms of animal ethical committee and
collection of adrenal glands was done immediately
after ½ h of sacrifice and placed in normal saline.
To avoid variations due to circadian rhythm, the
sacrifice time was kept constant between 10:30 and
11:30 AM. The fat and connective tissue around
the glands capsule were taken away carefully and
weighed.
Statistical analysis
The results are presented as MEAN±SEM of all
groups. The various parameters were subjected
to statistical analysis using GraphPad Prism
5 (version 5.03). Statistical significance was
determined by one-way ANOVA followed by
Dunnett’s multiple comparison test and “P” value
(P  0.05) being considered statistically significant.
RESULTS
Effect of noise stress on food, water, body
weight, and adrenal glands
The body weights of the EG1,
EG2
, EG3
, EG4
,
and EG5
rats were checked at the beginning of
the experiment no difference between their mean
body weights was observed (294.67 g. 293.22 g,
293.50 g, 294.80 g, and 293.82 g in rats). The effect
of varying intensities of noise stress for 1 h/30 day
onto the body weight of the experimental group
is depicted in Table 1 and Graph 1. Data showed
[Table 1] that, on 1st
day, no significant (P  0.05)
mean difference observed in the body weight of
EG1,
EG2
, EG3
, EG4
, and EG5
group when exposed
to noise stress of 60, 80, 100, 120, and 140 dB,
respectively [Graph 1], but after the 1st
day exposure
ofabovenoisestressasignificant(P0.05)increase
in food consumption [Table 2 and Graph 2a] and
highly significant (P  0.001) increased in water
consumption [Table 2 and Graph 2b] was noted
in all the experimental group compared to their
normal control group, respectively.
The consumption of food and water was decreased
in a highly significant (P  0.001) manner of EG1,
EG2
, EG3
, EG4
, and EG5
groups when compared
to their normal control group on 14th
day [Table 2
and Graph 2a and b] and also, a highly significant
Mehra, et al.: Effects of noise stress on male wistar rats
IJPBA/Apr-Jun-2020/Vol 11/Issue 2 94
(P  0.001) reduction was recorded in body weight
of EG3
, EG4
, and EG5
groups but no significant
(P  0.05) mean difference was recorded in the
weight of EG1
and EG2
groups of animals [Table 1
and Graph 1]. A weight gain was observed in EG1,
EG2
, and EG3
groups of animals and there mean
difference was not found statistically significant
(P  0.05), whereas a highly significant loss in
body weight was recorded in EG4
and EG5
groups
when compared to EG1
(60 dB) group on 30th
day,
respectively [Table 1 and Graph 1]. On the other
hand, 29th
day a recovered food consumption pattern
in EG1,
EG2
, and EG3
was found and the mean
difference of food consumption was not statistically
significant, conversely, the food consumption was
found to be highly significantly (P  0.001) reduced
in EG4
and EG5
groups when compared to their
normal control group. In contrast, animals of group
EG1
and EG2
showed a normal water consumption
as their water consumption mean value not found
significantly different (P  0.05) but EG3
group
showed a significant decrease (P  0.05) and
EG4
and EG5
group showed a highly significant
reduction in water consumption when compared to
Table 1: Effect of noise stress onto the body weight (g) of rats
Groups Noise treatment (1 h/30 days) Body weight (g) % decrease in body weight(g)
between 0th
and 30th
day
1st
day 15th
day 30th
day
EG1
60 dB 294.67±0.30 274.98±1.19*** 292.40±1.10ns
0.76
EG2
80 dB 293.22±0.46 272.22±1.98*** 292.12±1.20ns
0.37
EG3
100 dB 293.50±0.63 256.97±5.51*** 282.13±2.96ns
3.87
EG4
120 dB 294.80±0.28 236.00±2.17*** 218.47±4.87*** 25.89
EG5
140 dB 293.82±0.68 237.48±0.61*** 205.47±3.26*** 30.06
The values are expressed in Mean±SEM; (n=6). One-way ANOVA followed by Dunnett’s post-test; ***P0.001 versus 0th
days; ns
P0.05 versus 0th
day
Table 2: Effects of noise stress on the average total food and water consumption on 1st
, 14th
, and 29th
days
Group Noise stress Total food consumption (g) on Total water consumption (mL) on
1st
day 14th
day 29th
day 1st
day 14th
day 29th
day
Normal control - 128.80@
128.50#
129.10$
192.5a
193b
192.5c
EG1
60 dB 136.30@
* 65.90#
*** 110.00$ns
219.50a
*** 152.00b
*** 190.00c-ns
EG2
80 dB 136.10@
* 67.60#
*** 102.40$ns
218.00a
*** 151.00b
*** 183.00c-ns
EG3
100 dB 122.00@
* 54.50#
*** 80.40$ns
223.50a
*** 140.00b
*** 170.50c-
*
EG4
120 dB 125.70@
* 47.50#
*** 33.30$
*** 221.50a
*** 134.00b
*** 78.00c-
***
EG5
140 dB 117.30@
* 33.90#
*** 18.00$
*** 214.00a
*** 106.00b
*** 59.50c-
***
The values are expressed in Mean±SEM; (n=6). One-way ANOVA followed by Dunnett’s post-test; *P0.05 versus @
normal control; P0.001 versus #
normal control; ns
P0.05
versus $
normal control (of food consumption on 1st
, 15th
, and 30th
days), ***P0.001 versus a
normal control; ***P0.001 versus b
normal control; ns
P0.05 versus c
normal
control; *P0.05 versus c
normal control; ***
P0.001 versus c
normal control (of water consumption on 1st
, 14th
, and 29th
days)
Graph 1: Effect of noise stress onto the body weight. The values are expressed in Mean±SEM; (n = 6). One-way ANOVA
followed by Dunnett’s post-test; ***P  0.001 versus 1st
day; ns
P  0.05 versus 1st
day
Mehra, et al.: Effects of noise stress on male wistar rats
IJPBA/Apr-Jun-2020/Vol 11/Issue 2 95
their normal group on 29th
day, respectively [Table 2
and Graph 2a and b).
At the end of the study EG1,
EG2
, EG3
, EG4
, and EG5
group showed a percentage loss in body weight,
i.e., 0.76, 0.37, 3.87, 25.89, and 30.06% and in
contrast mean absolute weight of adrenal glands
[Graph 3] in EG1,
EG2
, EG3
, EG4
, and EG5
group
was increased, i.e., 0.05, 0.05, 0.09, 0.12, and 0.18
g when compared to their normal control group
(0.03 g) in the noise stress of 60, 80, 100, 120, and
140 dB for 1 h/day for 30 days, respectively.
DISCUSSION
Noisestressofvaryingsoundlevel(dB)wasutilized
as a tool to create noise stress and demonstrated its
effects on the food consumption behavior, water
consumption behavior, body weight, and absolute
adrenal glands weight of the 17-week old male
Wistar rats.
The present study shows that the noise stress of
60, 80, 100, 120, and 140 dB for 1 h daily to 30
days produced a minimum 0.76% (60 dB), 0.36%
(80 dB), and 3.87% (100 dB) to maximum 25.89%
(120 dB) and 30.06% (140 dB) percentage loss of
body weight in EG1,
EG2
, EG3
, EG4
, and EG5
group
of animals. The loss of body weight in animals
suggests some previously reported mechanisms
that stress could onset depressive episodes and it
activates the HPA axis resulting from regular short
but long-duration exposure to any stressful stimuli.
The stress has played a major role in altering the
PA axis proving that noise pollution is a stress
to which if exposed chronically will produce
alteration in the PA system thereby affecting the
various physiological and anatomical features
of the individual. In general, any level of stress
which is caused by a stimulus, definitely triggers
adrenocorticotropic hormone (ACTH) level, i.e.,
hypothalamo-hypophyseal-adrenocortical axis is
affected.[22]
The noise could be a hurtful stimulus
and does a negative impact on animals as well as
humans through many well-known mechanisms.
Like other stress, noise stress sets off neuronal and
Graph 2: (a) Effect of noise on food consumption behavior. The values are expressed in Mean±SEM; (n = 6). One-way
ANOVA followed by Dunnett’s post-test; *P  0.05 versus normal control; P  0.001 versus normal control; ns
P  0.05 versus
normal control. (b) Effect of noise on water consumption behavior ***P  0.001 versus normal control; ***P  0.001 versus
normal control; ns
P  0.05 versus normal control; *P  0.05 versus normal control; ***P  0.001 versus normal control
a
b
Mehra, et al.: Effects of noise stress on male wistar rats
IJPBA/Apr-Jun-2020/Vol 11/Issue 2 96
hormonal stress responses or it can cause severe
harmful impact through direct organ vibration.[9]
The present study’s observations showed that
there was a significant reduction in food and water
consumption behavior during 1st–14th days in all
experimental animals, when exposed to varying
repeated noise stress and it leads to a reduction in
body weight in the same duration so, the reduction
in weight loss could be associated with the reduced
intake of food and water during that specific
duration. Our results were correlated to the reported
finding of Hariram et al.[10]
and Sundar and Radha[22]
they have also reported that reduced food and water
consumption could be a reason for sudden weight
loss during noise stress burden. The reported work
concluded that a normal food and water consumption
behaviorofanimalscouldchangeifexposuretonoise
stress is about 80 dB,[10]
in our procedure, we exposed
experimentalanimalsnotonlyat80dBbutexposedat
lower to higher noise stress from 80 dB and observed
some interesting mechanism of adaptation at lower
noise stress from 80 dB. Although, on the 1st
day,
during noise exposure in all groups, food, and water
consumption was reduced, when noise exposure
was stopped in all groups, animals consume a large
volume of water and food which may be because
of inhibition and stimulation of ventromedial
hypothalamus and lateral hypothalamus due to the
pressure exerted in cortical areas of the brain[10]
but
when exposure was continued for 14 days some
observations were changed. Other research revealed
and confirm our study that reduced body fluids and
partially to a reduction in food consumption leads to
tissue catabolism.[23, 24]
In prolongation, when our study continued for
another 15 days with the same time duration and
exposure of noise stress it was observed that body
weight, food, and water consumption behavior
were recovered in some groups of animals, i.e.,
EG1
(60 dB),
EG2
(80 dB), and EG3
(100 dB) after
the 14th
–29th
days while exposure of noise stress
remains in continuous, alternatively, in during the
same time period, EG4
(120 dB) and EG5
(140 dB)
group of animals fails to regain body weight, food,
and water consumption behavior, this finding
suggests that when the animals exposed to the
same noise stress, the behavioral and physiological
impact of stress exposure are reduced, indicating
that the animals become adapted to the continued
noise stress. This process also termed habituation.
Quality, quantity, and duration can greatly influence
adaptation. Release ofACTH and prolactin appears
to be specific for the applied stressor so that the
potentiality of the PA axis and prolactin to respond
to a novel (heterotypic) stressor can be preserved.
This phenomenon has been termed as facilitation
and can be unmasked alternating stress.[25]
In present study, animals of group EG1
, EG2
,
and EG3
may follow “facilitation” and acquired
adaption after during 14th
–29th
days and defensive
against stress response, whereas other groups of
animals, i.e., EG4
and EG5
, which showed reduced
weight, food, and water consumption behavior may
not facilitate themselves and it is well known that
dysregulation of the HPA axis by frequent stress
or elevated levels of circulating cortisol produces
hyperactivity of the sympathetic adrenal-medullary
system, such as cortisol, corticosteroid-binding
globulin, and ACTH.[26]
Adaptation to continuous stress may further
slowdown the corticoadrenal response to the
stimulus and some neural circuits scheming
pituitary hormone secretion in response to a stress
stimulus are modified by frequent exposure. It
looks like there should be reallocation in the
sensitivity of the hormonal response to repeated
experience with a stress stimulus.[27]
In contrast,
animals were exposed to lower noise stress, i.e., 60,
80, and 100 1 h/day for 30 days, after 15 days they
were capable to adapt themselves comparatively to
those animals, received higher intensities of noise
Graph 3: Effect of noise on absolute adrenal weight. The
values are expressed in Mean±SEM; (n = 6). One-wayANOVA
followed by Dunnett’s post-test; ns
P  0.05 versus normal
control; **P  0.001 versus normal control; ***P  0.001
versus normal control (of adrenal weight on 30th
day)
Mehra, et al.: Effects of noise stress on male wistar rats
IJPBA/Apr-Jun-2020/Vol 11/Issue 2 97
level 120 and 140 dB fails to adapt themselves.
This could be due the mechanism discussed above
and it could be a factor of initiation of animal’s
homeostasis mechanism that helps animals by
changing hormonal pattern, as discussed above,
converselyfailingofhomeostasisin120and140dB
noise stress exposed animals could be a reason
to gradual reduction in body weight, food, and
water consumption behavior. It may be concluded
that higher noise stress exposure regularly for a
longer time period could affect the neurohumoral
apparatus and it suggests pathogenesis of some
diseases like hypertension and in a diseased state
feeding and water, consumption could suffer. In
support of this, we discussed a reported study that
revealed the role of catecholamine.
Stress is greater in the hypertensive than in the
normotensive subject and more catecholamines in
such stressed hypertensive subjects and production
of catecholamines give rise to a large variety
of symptoms and signs due to their effect on
hemodynamics and metabolism, including weight
loss even with normal appetite and food intake.[28,29]
Under different noise stress (conditions), the
absolute weight of adrenal glands was increased in
our observations. Experimental animals of the group
EG1
(60 dB and EG2
(80 dB) were not statistically
significant when compared to normal control but
the numerically absolute weight of the adrenal
glands was increased in respect to normal control,
whereas the absolute weight of adrenals in group
EG3
(100 dB), EG4
(120 dB), and EG5
(140 dB)
showed a highly significant increase in weight. This
confirms some previously reported work.
That stress induces adrenal‑medullary response
to release adrenaline which in turn stimulates β2
receptorsonthepituitarygland.Itinitiatestherelease
of ACTH that can excite the adrenal medulla and
cortex ensuing in the excess release of adrenaline
and corticosterone, respectively. The adrenal
hypertrophy (increase in adrenal gland weight)
takes place in response to the secretion of ACTH
from the pituitary for increased corticosterone from
cortical cells to contest stress[30]
and also continues
stress could alter the endocrine balance that disrupts
the establishment of the HPA axis, thus it leading
to inappropriate stress responses[31]
and also adrenal
medulla is alarmed due to rapid-onset responses to
stress challenges, consisting of an early release of
previously stored catecholamines which is directly
neurally mediated. By contrast, the adrenocortical
response to stress is delayed and consists of a
neuroendocrine cascade of humoral mediated
processes. It can thus be hypothesized that with a
short duration of noise stress (1 h), the adrenal
medulla response can be considered physiological,
and not yet implying morphological changes.[31]
CONCLUSION
In the light of above observation, it was concluded
that noise stress of varying level could affect the
normal physiological and psychological behavior of
the experimental animals. The results of the present
study showed that the effects of noise stress of
varyingsoundlevel(dB)onthebodyweight,feeding
and drinking behavior, and absolute weight of
adrenal glands of the male Wistar rats. Based on the
changes, it can be concluded that the body weight of
rats was decreased as feeding and drinking behavior
reduced by the effect of all applied noise stress
and body weight was increased in some group of
animals as their feeding and drinking were increased
while remaining in the same noise exposure till the
end of the study, this means there is some adaption
against the less noise stress whereas in high noise
stress some group of rats fails to adapt themselves.
Hence, it can be concluded longer duration
noise exposure of high noise stress can induce an
endocrine imbalance that disturbs the establishment
of the HPA axis, thereby, leading to abnormal stress
responses, whereas absolute adrenal weights of all
the experimental animals found increased and it can
also be concluded that there is a positive role of the
adrenal gland in stress management by producing
corticosterone animals. Further studies are needed
to identify the role of hormones which help animals
to acquired adaptation. The acute and chronic study
with respect to noise stress levels may be extended.
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Effects of Noise Stress on Body Weight and Adrenal Gland Weight of Male Wistar Rats

  • 1. © 2020, IJPBA. All Rights Reserved 91 Available Online at www.ijpba.info International Journal of Pharmaceutical Biological Archives 2020; 11(2):91-98 ISSN 2582 – 6050 RESEARCH ARTICLE Effects of Noise Stress on Body Weight and Adrenal Gland Weight of Male Wistar Rats Ravinder Kr Mehra1,2 *, Mahesh Prasad3 , Dinesh Kr Sharma4 , Ravinder Kr Mehra2 1 Department of Pharmacology, School of Pharmacy, Bharat Institute of Technology, Meerut, Uttar Pradesh, India, 2 Department of Pharmaceutical Sciences, IFTM University, Moradabad, Uttar Pradesh, India, 3 Department of Pharmacology, Kamla Nehru Institute of Management and Technology, Sultanpur, Uttar Pradesh, India, 4 Department of Pharmaceutical Sciences, Himalayan Institute of Pharmacy and Research, Uttarakhand, Uttar Pradesh, India. Received: 15 March 2020; Revised: 28 April 2020; Accepted: 10 May 2020 ABSTRACT The present study was designed to investigate the effects of varying intensities of noise stress onto the body and adrenal gland weight (absolute) of 17 weeks old (291–296) gram (g) Wistar rat. Animals were exposed to varying intensity of noise, i.e., 60 dB (Decibel), 80 dB, 100 dB, 120 dB, and 140 dB for 1 h (h)/day for 30 days. The dose of different noise intensities was delivered through a special fabricated noise chamber which had provided all daily needs of animal-like oxygen, food, and water. Food and water consumption behavior was recorded on 1st , 14th , and 29th days and body weight change recorded on 1st , 15th , and 30th days after the exposure of noise stress. Absolute adrenal glands weight was recorded on the 30th day. The results of the present study showed that the food, water consumption behavior, and body weight (g) of the experimental group EG1 (60 dB), EG2 (80 dB), EG3 (100 dB), EG4 (120 dB), and EG5 (140 dB) were found to be significantly (P 0.001) decreased during 1st –14th days and during 15th –30th days a recovery which were observed in food, water consumption, and in body weight of the EG1 (60 dB), EG2 (80 dB), and EG3 (100 dB). A percentage (%) decrease was found, i.e., 0.76, 0.37, 3.87, 25.89, and 30.06% in body weight of EG1 (60 dB), EG2 (80 dB), EG3 (100 dB), EG4 (120 dB), and EG5 (140 dB) experimental groups, respectively. Keywords: Body weight, decibel, noise stress INTRODUCTION In the current scenario, human and animal health affected by uncountable stimuli and these stimuli may responsible for the progression of several health issues. At present, we are surrounded by an environment that is filled with uncountable stressors. Stress can be defined as; it may a kind of physical, chemical, and emotional stimuli to which an individualfailstomakeasuitableadaptation.[1] Stress is a condition that aggravated by psychological, *Corresponding Author: Ravinder Kr Mehra, E-mail: rxravimehra@gmail.com physiological, or environmental stressors and could insult the homeostasis. It provokes the hypothalamic-pituitary-adrenal (HPA) axis and the sympathetic nervous system causing in a physiological change.[2] Our surrounding is layered with so many types of stress, from all others, noise stress intentionally (any loud concert) and non- intentionally (occupational) is regarded as the most common stimuli. Noise is regarded to every surplus sound generated by natural phenomena (e.g., wind, volcanic eruption, and oceans) or human-based sources (e.g., automobiles, machines, and explosions).[3,4] and environmental stress that is capable of causing hearing impairment, behavioral, mental, and physiological alterations.[5] Many epidemiological studies established a clear
  • 2. Mehra, et al.: Effects of noise stress on male wistar rats IJPBA/Apr-Jun-2020/Vol 11/Issue 2 92 connection between noise and vascular diseases including hypertension[6,7] and coronary heart disease.[8,9] Even, constant exposure to high levels of sound, for more than 5 years, may eventually lead noise-induced permanent threshold shift and can lead to elevated stress levels as well as stimulate violent behavior;[10] likewise, explosive noise may produce a massive adverse effect on the human body and mind.[11] Continuous noise stress can severely insult the homeostasis and this may initiate the events of cellular injury if the injury may stay continue due to persistent stress could open the way for several disease pathogenesis. It has been known that peptic ulcers occurred much morefrequentlyinwartimethaninpeacetime.[12] The study revealed that due to high levels of noise stress variety of health issues including cardiovascular diseases,[13] mental illness,[14] sleep disorder,[15] immune function,[16] hormonal levels,[17] respiratory systems,[18,19] and hypertension occurred. Furthermore, gastrointestinal motility disorder, gastritis, and peptic ulcers have been noticed in populations[2] because noise acts as a non-specific biological stressor and causes dysfunction of body organs and systems[20] and widespread disturbances at several levels in human organs and apparatus due to chemical and physiological modification of endocrine.[21] Noise stress of any kind is known to be a stress factor which alters some physiological and behavioral changes in human as well as animals. The present work is designed to study the effect of noise stress on animal body weight and the adrenal gland weight. MATERIALS AND METHODS Experimental animals 17 weeks old male Wistar rats weighing around 291–296 g were used for the present study. The research project was initiated after obtaining clearance from the Institutional Animal Ethical Committee of Kamla Nehru Institute of Management and Technology, Sultanpur, Uttar Pradesh, India. All procedures for housing and caring of animals were followed according to the guidelines of the ethical committee. Animals were acclimatized in normal animal house conditions for a week before starting the experimental protocol. Wistar rats were housed in an open wire cage in a temperature-controlled room (22–24°C) and 12 h light-dark cycle with free access to standard laboratory rat chow and tap water. To minimize all undesired stressors, such as handling, and habitat, animals were not exposed to noise for at least 1 week after delivery to the experimental room. Noise treatment Arectangular special noise chamber was fabricated to create a noise environment inside by fixing two adjacent speakers with amplifier and a noise meter was set to record and a controller knob was fixed to ensure the delivery of desired sound intensities in dB. A transparent acrylic front was provided to the noise chamber for viewing the activity of animals during experimentation. The chamber had all provisions for water, feed, and aeration. Before starting, the procedure animals were left inside the noise chamber without sound to get accustomed to the interior of the chamber during the acclimatization period. Study design Male Wistar rats of 17 weeks old (291–296 g) were included in this study. The animals were divided into five subgroups (n=6) as EG1 , EG2, EG3, EG4 , and EG5 based on noise stress exposure of for 1 h/day for 30 days, respectively. Every five groups were considered a statistical unit in this study in case of investigating effects of noise onto body weight. The time of noise stress exposure (9:00 AM–10: AM) was kept constant for each exposed group throughout the end of the experiment. Noise stress of 60, 80, 100, 120, and 140 dB intensities were delivered for 1 h/day for 30 day through a specially designed noise chamber to EG1 , EG2, EG3, EG4 , and EG5 experimental groups. To avoid age variation between all groups, an unbiased experimental protocol was decided to conduct and total study was conduct in total 150 days (30 days for each group) so, initially EG1 group was exposed to 60 dB noise stress for
  • 3. Mehra, et al.: Effects of noise stress on male wistar rats IJPBA/Apr-Jun-2020/Vol 11/Issue 2 93 1 h/day for 30 days and then the reaming EG2, EG3, EG4 , and EG5 groups were separately exposed to noise stress of 80, 100, 120, and 140 dB in 120 days, respectively. A normal control group was also constructed to evaluate the absolute weight of adrenal glands (g) and for comparing food and water consumption of EG1 , EG2, EG3, EG4 , and EG5 experimental groups, respectively, whereas in case of evaluating the effect of noise stress on the body weight in EG1 , EG2, EG3, EG4, and EG5 experimental groups, each group was considered a statistical unit and compared with EG1 (60 dB) group on 1st , 15th , and 30th days. Water and food intake observation Consumption of the food and water intake was recorded of the experimental animal group after delivery to the experimental room without access to noise stress and fed a homogeneous chow diet. A per-day analysis was performed to calculate food and water consumption in 1 week. During this acclimatization, per day food and water consumption was recorded. After the 1 week of acclimatization, animals have attained an age of 17 weeks and their average weight was found to be 291–296 g. Exposure to noise stress was started at the age of 17 weeks. On the 0th day, the initial body weight of each animal was recorded and exposure of noise stress started. A per-day analysis of food and water consumption behavior was recorded to evaluate the effect of noise stress onto the same but final interpretation was done with results of 1st , 14th , and 29th days consumption. Measurement of body weight and adrenals weight Weight measurement was done on 1st , 15th , and 30th days with the help of an electronic weighing scale of the group under experimentation. Exposure of noise was started on the 1st day after recording the initial body weight. On the 29th day, all animals were deprived of food and had to access only water in their habitat. On the 30th day, after recording the final weight, animals were sacrificed as per the norms of animal ethical committee and collection of adrenal glands was done immediately after ½ h of sacrifice and placed in normal saline. To avoid variations due to circadian rhythm, the sacrifice time was kept constant between 10:30 and 11:30 AM. The fat and connective tissue around the glands capsule were taken away carefully and weighed. Statistical analysis The results are presented as MEAN±SEM of all groups. The various parameters were subjected to statistical analysis using GraphPad Prism 5 (version 5.03). Statistical significance was determined by one-way ANOVA followed by Dunnett’s multiple comparison test and “P” value (P 0.05) being considered statistically significant. RESULTS Effect of noise stress on food, water, body weight, and adrenal glands The body weights of the EG1, EG2 , EG3 , EG4 , and EG5 rats were checked at the beginning of the experiment no difference between their mean body weights was observed (294.67 g. 293.22 g, 293.50 g, 294.80 g, and 293.82 g in rats). The effect of varying intensities of noise stress for 1 h/30 day onto the body weight of the experimental group is depicted in Table 1 and Graph 1. Data showed [Table 1] that, on 1st day, no significant (P 0.05) mean difference observed in the body weight of EG1, EG2 , EG3 , EG4 , and EG5 group when exposed to noise stress of 60, 80, 100, 120, and 140 dB, respectively [Graph 1], but after the 1st day exposure ofabovenoisestressasignificant(P0.05)increase in food consumption [Table 2 and Graph 2a] and highly significant (P 0.001) increased in water consumption [Table 2 and Graph 2b] was noted in all the experimental group compared to their normal control group, respectively. The consumption of food and water was decreased in a highly significant (P 0.001) manner of EG1, EG2 , EG3 , EG4 , and EG5 groups when compared to their normal control group on 14th day [Table 2 and Graph 2a and b] and also, a highly significant
  • 4. Mehra, et al.: Effects of noise stress on male wistar rats IJPBA/Apr-Jun-2020/Vol 11/Issue 2 94 (P 0.001) reduction was recorded in body weight of EG3 , EG4 , and EG5 groups but no significant (P 0.05) mean difference was recorded in the weight of EG1 and EG2 groups of animals [Table 1 and Graph 1]. A weight gain was observed in EG1, EG2 , and EG3 groups of animals and there mean difference was not found statistically significant (P 0.05), whereas a highly significant loss in body weight was recorded in EG4 and EG5 groups when compared to EG1 (60 dB) group on 30th day, respectively [Table 1 and Graph 1]. On the other hand, 29th day a recovered food consumption pattern in EG1, EG2 , and EG3 was found and the mean difference of food consumption was not statistically significant, conversely, the food consumption was found to be highly significantly (P 0.001) reduced in EG4 and EG5 groups when compared to their normal control group. In contrast, animals of group EG1 and EG2 showed a normal water consumption as their water consumption mean value not found significantly different (P 0.05) but EG3 group showed a significant decrease (P 0.05) and EG4 and EG5 group showed a highly significant reduction in water consumption when compared to Table 1: Effect of noise stress onto the body weight (g) of rats Groups Noise treatment (1 h/30 days) Body weight (g) % decrease in body weight(g) between 0th and 30th day 1st day 15th day 30th day EG1 60 dB 294.67±0.30 274.98±1.19*** 292.40±1.10ns 0.76 EG2 80 dB 293.22±0.46 272.22±1.98*** 292.12±1.20ns 0.37 EG3 100 dB 293.50±0.63 256.97±5.51*** 282.13±2.96ns 3.87 EG4 120 dB 294.80±0.28 236.00±2.17*** 218.47±4.87*** 25.89 EG5 140 dB 293.82±0.68 237.48±0.61*** 205.47±3.26*** 30.06 The values are expressed in Mean±SEM; (n=6). One-way ANOVA followed by Dunnett’s post-test; ***P0.001 versus 0th days; ns P0.05 versus 0th day Table 2: Effects of noise stress on the average total food and water consumption on 1st , 14th , and 29th days Group Noise stress Total food consumption (g) on Total water consumption (mL) on 1st day 14th day 29th day 1st day 14th day 29th day Normal control - 128.80@ 128.50# 129.10$ 192.5a 193b 192.5c EG1 60 dB 136.30@ * 65.90# *** 110.00$ns 219.50a *** 152.00b *** 190.00c-ns EG2 80 dB 136.10@ * 67.60# *** 102.40$ns 218.00a *** 151.00b *** 183.00c-ns EG3 100 dB 122.00@ * 54.50# *** 80.40$ns 223.50a *** 140.00b *** 170.50c- * EG4 120 dB 125.70@ * 47.50# *** 33.30$ *** 221.50a *** 134.00b *** 78.00c- *** EG5 140 dB 117.30@ * 33.90# *** 18.00$ *** 214.00a *** 106.00b *** 59.50c- *** The values are expressed in Mean±SEM; (n=6). One-way ANOVA followed by Dunnett’s post-test; *P0.05 versus @ normal control; P0.001 versus # normal control; ns P0.05 versus $ normal control (of food consumption on 1st , 15th , and 30th days), ***P0.001 versus a normal control; ***P0.001 versus b normal control; ns P0.05 versus c normal control; *P0.05 versus c normal control; *** P0.001 versus c normal control (of water consumption on 1st , 14th , and 29th days) Graph 1: Effect of noise stress onto the body weight. The values are expressed in Mean±SEM; (n = 6). One-way ANOVA followed by Dunnett’s post-test; ***P 0.001 versus 1st day; ns P 0.05 versus 1st day
  • 5. Mehra, et al.: Effects of noise stress on male wistar rats IJPBA/Apr-Jun-2020/Vol 11/Issue 2 95 their normal group on 29th day, respectively [Table 2 and Graph 2a and b). At the end of the study EG1, EG2 , EG3 , EG4 , and EG5 group showed a percentage loss in body weight, i.e., 0.76, 0.37, 3.87, 25.89, and 30.06% and in contrast mean absolute weight of adrenal glands [Graph 3] in EG1, EG2 , EG3 , EG4 , and EG5 group was increased, i.e., 0.05, 0.05, 0.09, 0.12, and 0.18 g when compared to their normal control group (0.03 g) in the noise stress of 60, 80, 100, 120, and 140 dB for 1 h/day for 30 days, respectively. DISCUSSION Noisestressofvaryingsoundlevel(dB)wasutilized as a tool to create noise stress and demonstrated its effects on the food consumption behavior, water consumption behavior, body weight, and absolute adrenal glands weight of the 17-week old male Wistar rats. The present study shows that the noise stress of 60, 80, 100, 120, and 140 dB for 1 h daily to 30 days produced a minimum 0.76% (60 dB), 0.36% (80 dB), and 3.87% (100 dB) to maximum 25.89% (120 dB) and 30.06% (140 dB) percentage loss of body weight in EG1, EG2 , EG3 , EG4 , and EG5 group of animals. The loss of body weight in animals suggests some previously reported mechanisms that stress could onset depressive episodes and it activates the HPA axis resulting from regular short but long-duration exposure to any stressful stimuli. The stress has played a major role in altering the PA axis proving that noise pollution is a stress to which if exposed chronically will produce alteration in the PA system thereby affecting the various physiological and anatomical features of the individual. In general, any level of stress which is caused by a stimulus, definitely triggers adrenocorticotropic hormone (ACTH) level, i.e., hypothalamo-hypophyseal-adrenocortical axis is affected.[22] The noise could be a hurtful stimulus and does a negative impact on animals as well as humans through many well-known mechanisms. Like other stress, noise stress sets off neuronal and Graph 2: (a) Effect of noise on food consumption behavior. The values are expressed in Mean±SEM; (n = 6). One-way ANOVA followed by Dunnett’s post-test; *P 0.05 versus normal control; P 0.001 versus normal control; ns P 0.05 versus normal control. (b) Effect of noise on water consumption behavior ***P 0.001 versus normal control; ***P 0.001 versus normal control; ns P 0.05 versus normal control; *P 0.05 versus normal control; ***P 0.001 versus normal control a b
  • 6. Mehra, et al.: Effects of noise stress on male wistar rats IJPBA/Apr-Jun-2020/Vol 11/Issue 2 96 hormonal stress responses or it can cause severe harmful impact through direct organ vibration.[9] The present study’s observations showed that there was a significant reduction in food and water consumption behavior during 1st–14th days in all experimental animals, when exposed to varying repeated noise stress and it leads to a reduction in body weight in the same duration so, the reduction in weight loss could be associated with the reduced intake of food and water during that specific duration. Our results were correlated to the reported finding of Hariram et al.[10] and Sundar and Radha[22] they have also reported that reduced food and water consumption could be a reason for sudden weight loss during noise stress burden. The reported work concluded that a normal food and water consumption behaviorofanimalscouldchangeifexposuretonoise stress is about 80 dB,[10] in our procedure, we exposed experimentalanimalsnotonlyat80dBbutexposedat lower to higher noise stress from 80 dB and observed some interesting mechanism of adaptation at lower noise stress from 80 dB. Although, on the 1st day, during noise exposure in all groups, food, and water consumption was reduced, when noise exposure was stopped in all groups, animals consume a large volume of water and food which may be because of inhibition and stimulation of ventromedial hypothalamus and lateral hypothalamus due to the pressure exerted in cortical areas of the brain[10] but when exposure was continued for 14 days some observations were changed. Other research revealed and confirm our study that reduced body fluids and partially to a reduction in food consumption leads to tissue catabolism.[23, 24] In prolongation, when our study continued for another 15 days with the same time duration and exposure of noise stress it was observed that body weight, food, and water consumption behavior were recovered in some groups of animals, i.e., EG1 (60 dB), EG2 (80 dB), and EG3 (100 dB) after the 14th –29th days while exposure of noise stress remains in continuous, alternatively, in during the same time period, EG4 (120 dB) and EG5 (140 dB) group of animals fails to regain body weight, food, and water consumption behavior, this finding suggests that when the animals exposed to the same noise stress, the behavioral and physiological impact of stress exposure are reduced, indicating that the animals become adapted to the continued noise stress. This process also termed habituation. Quality, quantity, and duration can greatly influence adaptation. Release ofACTH and prolactin appears to be specific for the applied stressor so that the potentiality of the PA axis and prolactin to respond to a novel (heterotypic) stressor can be preserved. This phenomenon has been termed as facilitation and can be unmasked alternating stress.[25] In present study, animals of group EG1 , EG2 , and EG3 may follow “facilitation” and acquired adaption after during 14th –29th days and defensive against stress response, whereas other groups of animals, i.e., EG4 and EG5 , which showed reduced weight, food, and water consumption behavior may not facilitate themselves and it is well known that dysregulation of the HPA axis by frequent stress or elevated levels of circulating cortisol produces hyperactivity of the sympathetic adrenal-medullary system, such as cortisol, corticosteroid-binding globulin, and ACTH.[26] Adaptation to continuous stress may further slowdown the corticoadrenal response to the stimulus and some neural circuits scheming pituitary hormone secretion in response to a stress stimulus are modified by frequent exposure. It looks like there should be reallocation in the sensitivity of the hormonal response to repeated experience with a stress stimulus.[27] In contrast, animals were exposed to lower noise stress, i.e., 60, 80, and 100 1 h/day for 30 days, after 15 days they were capable to adapt themselves comparatively to those animals, received higher intensities of noise Graph 3: Effect of noise on absolute adrenal weight. The values are expressed in Mean±SEM; (n = 6). One-wayANOVA followed by Dunnett’s post-test; ns P 0.05 versus normal control; **P 0.001 versus normal control; ***P 0.001 versus normal control (of adrenal weight on 30th day)
  • 7. Mehra, et al.: Effects of noise stress on male wistar rats IJPBA/Apr-Jun-2020/Vol 11/Issue 2 97 level 120 and 140 dB fails to adapt themselves. This could be due the mechanism discussed above and it could be a factor of initiation of animal’s homeostasis mechanism that helps animals by changing hormonal pattern, as discussed above, converselyfailingofhomeostasisin120and140dB noise stress exposed animals could be a reason to gradual reduction in body weight, food, and water consumption behavior. It may be concluded that higher noise stress exposure regularly for a longer time period could affect the neurohumoral apparatus and it suggests pathogenesis of some diseases like hypertension and in a diseased state feeding and water, consumption could suffer. In support of this, we discussed a reported study that revealed the role of catecholamine. Stress is greater in the hypertensive than in the normotensive subject and more catecholamines in such stressed hypertensive subjects and production of catecholamines give rise to a large variety of symptoms and signs due to their effect on hemodynamics and metabolism, including weight loss even with normal appetite and food intake.[28,29] Under different noise stress (conditions), the absolute weight of adrenal glands was increased in our observations. Experimental animals of the group EG1 (60 dB and EG2 (80 dB) were not statistically significant when compared to normal control but the numerically absolute weight of the adrenal glands was increased in respect to normal control, whereas the absolute weight of adrenals in group EG3 (100 dB), EG4 (120 dB), and EG5 (140 dB) showed a highly significant increase in weight. This confirms some previously reported work. That stress induces adrenal‑medullary response to release adrenaline which in turn stimulates β2 receptorsonthepituitarygland.Itinitiatestherelease of ACTH that can excite the adrenal medulla and cortex ensuing in the excess release of adrenaline and corticosterone, respectively. The adrenal hypertrophy (increase in adrenal gland weight) takes place in response to the secretion of ACTH from the pituitary for increased corticosterone from cortical cells to contest stress[30] and also continues stress could alter the endocrine balance that disrupts the establishment of the HPA axis, thus it leading to inappropriate stress responses[31] and also adrenal medulla is alarmed due to rapid-onset responses to stress challenges, consisting of an early release of previously stored catecholamines which is directly neurally mediated. By contrast, the adrenocortical response to stress is delayed and consists of a neuroendocrine cascade of humoral mediated processes. It can thus be hypothesized that with a short duration of noise stress (1 h), the adrenal medulla response can be considered physiological, and not yet implying morphological changes.[31] CONCLUSION In the light of above observation, it was concluded that noise stress of varying level could affect the normal physiological and psychological behavior of the experimental animals. The results of the present study showed that the effects of noise stress of varyingsoundlevel(dB)onthebodyweight,feeding and drinking behavior, and absolute weight of adrenal glands of the male Wistar rats. Based on the changes, it can be concluded that the body weight of rats was decreased as feeding and drinking behavior reduced by the effect of all applied noise stress and body weight was increased in some group of animals as their feeding and drinking were increased while remaining in the same noise exposure till the end of the study, this means there is some adaption against the less noise stress whereas in high noise stress some group of rats fails to adapt themselves. Hence, it can be concluded longer duration noise exposure of high noise stress can induce an endocrine imbalance that disturbs the establishment of the HPA axis, thereby, leading to abnormal stress responses, whereas absolute adrenal weights of all the experimental animals found increased and it can also be concluded that there is a positive role of the adrenal gland in stress management by producing corticosterone animals. Further studies are needed to identify the role of hormones which help animals to acquired adaptation. The acute and chronic study with respect to noise stress levels may be extended. REFERENCES 1. Jameel MK, Joshi AR, Dawane J, Padwal M, Joshi AR, Pandit VA, Melinkeri RR. Effectof various physical stress models on serum cortisol level in wistar rats. J
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