The Effect of Epinephrine Administration on the Level of

Gonadotropin Hormones of Japan Strain Female Mice (Mus

Musculus)

Aulia Asman*, Debby Sinthania, and Linda Marni

Nursing Diploma, Padang State University, Padang, West Sumatera, Indonesia

Wisma Indah Enam Blok N/6 Balai Baru, Kec. Kuranji, Kota Padang, Sumatera Barat 25155

Keywords: Epinephrine, FSH, LH.

Abstract: Physical, chemical, and psychological stressors can affect the pulsatile frequency and amplitude of GnRH;

this is important for FSH and LH secretion. Excessive increase in pulsation can reduce and stop FSH and

LH secretion. This study aimed to prove the effect of epinephrine to FSH and LH levels. This study was an

experimental laboratory with a post-test only control group design. This study used 24 female mice (Mus

musculus) consisting of 6 groups, namely 1 control group and 5 treatment groups based on differences in the

administration of epinephrine of 0.001 mg/ml, 0.002 mg/ml, 0.003 mg/ml, 0.004 mg/ml, and 0.005 mg/ml

applied every day for 20 days starting at the beginning of the pro-estrus cycle. The results were analyzed

using the One-Way ANOVA continued Multiple Comparisons Bonferroni test. The results showed that the

dose difference in the administration of epinephrine gave a significant difference (p <0.05) on FSH and LH

levels starting at 0.001 mg/ml and 0.002 mg/ml, 0.003 mg/ml, 0.004 mg/ml, 0.005 mg/ml with control. It

can be concluded that chemical stressors (epinephrine) can reduce FSH and LH levels. It is recommended

that further research adds measurements on the levels of estrogen and progesterone and the levels of

catecholamine.

INTRODUCTION

During their life, human beings can always

experience stress. If stress continues, it will cause

interference with various body systems, one of

which is the reproductive system. The function of

the gonadal axis can change under certain conditions

such as physical, chemical, and psychological

stressors. This stressor can cause an imbalance in the

hypothalamus – pituitary - ovarian axis. The

reproductive system imbalance can be in the form of

ovulation disorders or suppression. Reproductive

disorders that occur can be menstrual disorders,

which include menarche delay, a short and

inadequate luteal phase, and even secondary

amenorrhea; these can cause reversible infertility

(Vander Borght and Wyns, 2018).

One of the reproductive disorders in women is

hormonal disorders. Hormonal disorders can cause

disruption in the process of development and

formation of egg cells (ovum) through the process of

oogenesis. Oogenesis occurs in the ovary through

certain stages and is controlled by hormones,

especially gonadotropin Follicle Stimulating

Hormone (FSH) and Luteinizing Hormone (LH).

This gonadotropin hormone is produced by the

anterior pituitary gland through Gonadotropin

stimulation Releasing Hormone (GnRH) from the

hypothalamus (Barret et al., 2012; Klein, 2014)

Stressors, be it physical, chemical, and

psychological, can activate the sympathetic nervous

system and adrenal response (Tanner, Sport and

Gore, 2012). Activation of the sympathetic nervous

system by stressors can cause the release of local

norepinephrine (NE) neurotransmitters at

postganglionic sympathetic nerve endings, while the

sympathetic nervous system will stimulate the

adrenal medulla so epinephrine will be released into

the circulation (Bonert and Melmed, 2017).

Epinephrine has a unique characteristic to modulate

norepinephrine (NE), where the norepinephrine

released will be duplicated and strengthened by

epinephrine to reach the same place through

circulation (Barret et al., 2012; Klein, 2014)

Asman, A., Sinthania, D. and Marni, L.

The Effect of Epinephrine Administration on the Level of Gonadotropin Hormones of Japan Strain Female Mice (Mus Musculus).

DOI: 10.5220/0009124301190123

In Proceedings of the 2nd Health Science International Conference (HSIC 2019), pages 119-123

ISBN: 978-989-758-462-6

119

Stress can increase the production of glands or

the epinephrine hormone. Under normal

circumstances, hormones can have a positive effect,

such as making us more motivated to work or make

us more focused. However, excessive hormone

production due to prolonged stress will lead to

fatigue and even depression. Physical illness will

also present easily, as a result of faster blood

pumping, which disrupts the metabolism and the

oxidation process in the body. The epinephrine

hormone is synthesized in the medulla adrenal gland

by chromatin cells (Gardner and Shobac, 2011)

Continuous stress can affect the frequency and

amplitude of pulses of Gonadotropin-Releasing

Hormone (GnRH), which is important for the

secretion of Follicle Stimulating Hormone (FSH)

and Luteinizing Hormone (LH). In addition,

stressors can also activate the sympathetic nervous

system (release of norepinephrine) and the adrenal

response (release of epinephrine). Increased levels of

epinephrine and norepinephrine can increase the

GnRH pulse. Excessive increase in pulsation can

reduce and stop FSH and LH secretion. This

reduction in FSH and LH will disrupt the oogenesis

process (Gardner and Shobac, 2011; Hall and

Guyton, 2011).

Normally GnRH is secreted through episodic

pulses, which is important for normal FSH and LH

secretions (Barret et al., 2012). Studies show that

changes in FSH and LH secretion require the

pulsatile release of GnRH with a frequency of

amplitude within the critical limit. This has been

proven by research on monkeys who were

administered 1 microgram of GnRH/minute for each

hour (1 pulsation/hour)—the treatment results in ± 2

micrograms/ml concentration of GnRH in human

portal blood. Increasing the frequency of GnRH

pulses to 2 and 5 pulses/hour will stop gonadotropin

secretion. Gonadotropin secretion will also decrease

if the dose of GnRH is increased (Arief, 2006).

Experiments on female mice, by giving sub-

cutaneous epinephrine at 0.001 mg can affect the

pulsatile frequency and amplitude of GnRH, thereby

disrupting FSH and LH production, which results in

a decrease in the number of tertiary and deGraaf

follicles in the ovary (Gardner and Shobac, 2011).

Experiments on male mice by giving sub-cutaneous

epinephrine at 0.001 mg, 0.005 mg, and 0.01 mg

causes disruption in the process of spermatogenesis

due to disruption in the secretion of GnRH in

stimulating the production of FSH and LH (Arief,

2006)

Another study confirms the administration of

epinephrine at 0.002 mg continuously for one cycle

of spermatogenesis in mice (Mus musculus) reduces

the quantity and quality of spermatozoa (Abdullah,

2008). Another study uses epinephrine at 0.001 mg,

0.005 mg, and 0.01 mg; the results show that

administration at 0.001 mg alone has significantly

reduced the number of spermatogenic cells (Utami,

2010). This is because of the disruption of hormonal

arrangements in the hypothalamic-pituitary-

testicular axis pathway.

According to a study in fertility and sterility,

scientists from the University of California have

discovered the negative effects of stress on the

possibility of pregnancy on women. The stress felt

by participants in the study turned out to affect the

number of eggs and embryos produced—too much

stress means fewer eggs produced and fertilized.

Women who are easily offended, angry, and

depressed have fewer eggs (Aggarwal et al., 2013).

Based on the aforementioned description, this

study was conducted to determine the effect of

epinephrine on gonadotropin levels of female mice

(Mus musculus).

METHODS

The research was a laboratory experiment using a

post-test only control group. Treatment of

experimental animals was done at the Pharmacy

Laboratory, Faculty of Pharmacy of Andalas

University, Padang. Examination of FSH and LH

hormone levels was conducted at the Laboratory of

Biochemistry, Faculty of Medicine of Andalas

University, Padang. The research was conducted for

six months. The population of this study was female

mice (Mus musculus) from the Pharmacy

Laboratory, Faculty of Pharmacy of Andalas

University, Padang. The study needed 24 mice, yet

due to the possibility of death, we had five mice in

each of the six groups, so we kept 30 mice in total.

RESULTS AND DISCUSSION

The level of FSH hormone on female mice (mus

musculus) of the control group and treatment

groups at the proestrus phase.

Table 1 shows that the average FSH hormone level

in the control and treatment groups decreases—the

more epinephrine administered, the smaller the

average is. The normality test (Kolmogorov

Smirnov) shows that data was normally distributed

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so parametric tests (ANOVA) could be done. Based

on the results of the statistical test, there was a

significant difference between the control and

treatment groups (p <0.05). Therefore, the statistical

test continued with the Multiple Comparisons (post

hoc test) employing the Bonferroni test type and the

results are presented in Table 2.

Table 2 shows that the average between the

control group and the treatment group is different.

The higher the dose of epinephrine administered, the

bigger the difference in the average is. The

difference was significant starting with epinephrine

dose of 0.001 mg (p<0.05). The administration of

epinephrine of 0.002 mg, 0.003 mg, 0.004 mg, and

0.005 mg also gave a significant difference.

Table 1: The level of FSH hormone on female mice (Mus musculus) of the control group and treatment groups at the

proestrus phase.

Treatment

Repetition Average

(mIU/ml)

1 2 3 4

K [Control] 0.720 0.736 0.776 0.844 0.769 0.055

P1 [0.001] 0.332 0.342 0.360 0.378 0.353 0.020

P2 [0.002] 0.390 0.308 0.322 0.342 0.340 0.035

P3 [0.003] 0.300 0.322 0.360 0.360 0.335 0.029

P4 [0.004] 0.368 0.342 0.262 0.322 0.323 0.045

P5 [0.005] 0.280 0.332 0.368 0.308 0.322 0.037

Table 2: The Results of the multiple comparisons of FSH hormone levels between the control and treatment groups.

Control Epinephrine Dose (mg)

Average

Difference

(mIU/ml )

p-value

Control [0.001] 0.416 0.000

[0.002] 0.428 0.000

[0.003] 0. 433 0.000

[0.004] 0.445 0.000

[0.005] 0.447 0.000

Table 3: The level of LH hormone on female mice (Mus musculus) of the control group and treatment groups at the

proestrus phase.

Treatment

Repetition

Average

(mIU/ml)

1 2 3 4

K [Control] 0.304 0.354 0.364 0.392 0.353 0.036

P1 [0.001] 0.198 0.158 0.182 0.218 0.189 0.025

P2 [0.002] 0.158 0.182 0.182 0.198 0.180 0.016

P3 [0.003] 0.170 0.190 0.170 0.182 0.178 0.009

P4 [0.004] 0.158 0.172 0.170 0.182 0.170 0.009

P5 [0.005] 0.135 0.182 0.140 0.182 0.159 0.025

Table 4: The results of the multiple comparisons of LH hormone levels between the control and treatment groups.

Control Epinephrine Dose (mg)

Average

Difference

(mIU/ml )

p-value

Control [0.001] 0.164 0.000

[0.002] 0.173 0.000

[0.003] 0. 175 0.000

[0.004] 0.183 0.000

[0.005] 0.193 0.000

The Effect of Epinephrine Administration on the Level of Gonadotropin Hormones of Japan Strain Female Mice (Mus Musculus)

121

The level of LH hormone on female mice (Mus

musculus) of the control group and treatment

groups at the proestrus phase.

Table 3 shows that the average LH hormone level in

the control and treatment groups decreases—the

more epinephrine administered, the smaller the

average is.

The normality test (Kolmogorov Smirnov) shows

that data was normally distributed so parametric

tests (ANOVA) could be done. Based on the results

of the statistical test, there was a significant

difference between the control and treatment groups

(p <0.05). Therefore, the statistical test continued

with the Multiple Comparisons (post hoc test)

employing the Bonferroni test type and the results

are presented in Table 4.

Table 4 shows that the average between the

control group and the treatment group is different.

The higher the dose of epinephrine administered, the

bigger the difference in the average is. The

difference was significant starting with epinephrine

dose of 0.001 mg (p<0.05). The administration of

epinephrine of 0.002 mg, 0.003 mg, 0.004 mg, and

0.005 mg also gave a significant difference.

The effect of epinephrine administration on FSH

hormones levels.

The results of the study confirm that the FSH

hormone levels of mice decreased. The higher the

dose of epinephrine administered, the lower the FSH

hormone level produced was. This shows that the

administration of epinephrine with a dose of

0.001 mg has begun to influence the FSH hormone

levels. The administration of epinephrine at 0.002

mg, 0.003 mg, 0.004 mg, and 0.005 mg also

influenced the FSH hormone levels.

The effect of epinephrine administration on LH

hormones levels.

The results of the study confirm that the LH

hormone levels of mice decreased. The higher the

dose of epinephrine administered, the lower the LH

hormone level produced was. This shows that the

administration of epinephrine with a dose of 0.001

mg has begun to influence the LH hormone levels.

The administration of epinephrine at 0.002 mg,

0.003 mg, 0.004 mg, and 0.005 mg also influenced

the LH hormone levels.

This means that the administration of stressors

using low epinephrine levels has caused pulsatile

changes in GnRH in the hypothalamus that exceeds

the critical limit; this results resulting in a significant

decrease in GnRH production leading to down-

regulation of the anterior pituitary that finally causes

a decrease in FSH and LH secretion (Guyton and

Hall, 2014).

Repeated epinephrine administration as stressors

during the study caused the activation of the

sympathetic nervous system, which could cause the

release of local norepinephrine neurotransmitters at

the postganglionic sympathetic nerve end, while the

sympathetic nervous system would stimulate the

adrenal medulla so epinephrine would be released

into the circulation. GnRH pulsation control can be

influenced by catecholaminergic (epinephrine and

norepinephrine). Catecholamine may be working by

changing the frequency (and possibly amplitude) of

GnRH release. Increasing the pulse frequency and

amplitude of GnRH can reduce and stop

gonadotropin secretion. Finally, it will suppress FSH

and LH secretion (Hall and Guyton, 2011; Barret et

al., 2012).

Repeated and prolonged epinephrine

administration will also cause activation of the

amygdala in the limbic system. This system will

stimulate the release of hormones from the

hypothalamus, namely Corticotropic Releasing

Hormone (CRH). This hormone will directly inhibit

hypothalamic GnRH secretion. The result is reduced

stimulation to the anterior pituitary leading to

reduced FSH and LH secretion (Hall and Guyton,

2011).

Hormonal regulation of female reproductive

function occurs in a pathway called the

hypothalamus-pituitary-ovarian axis. This regulatory

mechanism starts from the hypothalamus gland with

the release of GnRH. Then, GnRH will stimulate the

anterior pituitary gland and the stimulation will

make the anterior pituitary to release the FSH and

LH hormone. FSH and LH hormone will then

stimulate the ovaries to produce estrogen and

progesterone (Barret et al., 2012).

CONCLUSIONS

Stressor using epinephrine administered repeatedly

will affect the pulsatile frequency and amplitude of

GnRH, which is important for FSH and LH

secretion. Research suggests that repeated

administration of subcutaneous epinephrine at 0.001

mg causes a decrease in the number of tertiary and

deGraaf follicles in the ovary of mice caused by

disruption of the hypothalamic-pituitary- ovary axis

(Gusty, 2007). Other study also confirms that the

administration of epinephrine at a dose of 0.001 mg

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has reduced the number of primary follicles in the

ovary of mice (Utami, 2010).

One study confirms that the administration of

epinephrine at 0.002 mg continuously for 1 cycle of

spermatogenesis in mice (Mus musculus) reduces the

quantity and quality of spermatozoa (Abdullah,

2008). Another study uses epinephrine at 0.001 mg,

0.005 mg, and 0.01 mg; the results show that

administration at 0.001 mg alone has significantly

reduced the number of spermatogenic cells (Trussell,

Kunselman and Legro, 2010). This is because of the

disruption of hormonal arrangements in the

hypothalamic-pituitary- testicular axis pathway.

The results of this study are also in accordance

with the research conducted by (Arief, 2006), where

the administration of epinephrine at 0.001 mg and

0.005 mg and 0.01 mg causes a significant effect on

the process of spermatogenesis. The stressors given

affect the frequency and amplitude of the

hypothalamus so the regulation of hormonal

secretions in the hypothalamic-pituitary-testicular

axis does not occur harmoniously.

ACKNOWLEDGEMENTS

The authors thanks Aulia Asman, Debby Sinthania,

and Linda Marni for their dedicated work in

collecting data used in this article as a part of the

objective of our research for the effect of

epinephrine administaration on the level of

gonadotropin hormones of Japan strain female mice

(Mus Musculus).

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The Effect of Epinephrine Administration on the Level of Gonadotropin Hormones of Japan Strain Female Mice (Mus Musculus)

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