Acupuncture Can Increase Spermatogenic Cells in Albino Rats

(Rattus norvegicus) Exposed by Heatstroke

Muhammad Thohawi Elziyad Purnama

, Imas Hapsari Rahmaningtyas

, Ainun Septia Putri

, Angela

Swasti Ivana Lee

, Fachrun Nisa Tatimma

and Herlina Masyitoh

Department of Veterinary Anatomy, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, Indonesia

Department of Veterinary Reproduction, Faculty of Veterinary Medicine, Universitas Airlangga, Surabaya, Indonesia

Keywords: Acupuncture, heatstroke, spermatogenic cells, testosterone.

Abstract: Global warming is the newest condition that causes many re-emerging diseases. Heatstroke has been

predicted as one of the global warming effects that made an important role in reproductive disease.

Heatstroke with a high temperature in environmental conditions can potentially cause infertility, especially

in physiological functions of the reproductive system. The objective of this study is to examine the potential

of acupuncture in increasing spermatogenic cells and testosterone levels. Acupuncture therapy is expected

to be an alternative solution for reducing body temperature and restoring the function of the reproductive

system. In this study, 20 albino rats were used and divided into five treatments, wherein negative control

group (C-) was not exposed to heatstroke and acupuncture and positive control group (C+) was exposed to

heatstroke for 20 seconds. Treatment groups (T1, T2, T3) were exposed to heatstroke and acupuncture for 5;

10; and 20 seconds/rat/day. The acupuncture treatment has been performed on Wei-Jian and Ming-Men

points for one week. The test was prepared with hematoxylin-eosin staining to evaluate spermatogenic cells.

The blood serum collected for assay was standardized by testosterone ELISA kit. The data of spermatogenic

cells and testosterone levels were analyzed using ANOVA, followed by Duncan test when there were

significantly different results (p<0.05). The results showed that the spermatogenic cells of (C+) were

significantly different (p<0.05) from those of T1, T2, and T3 and the testosterone levels of the treatment

groups were not significantly different (p>0.05). It can be concluded that acupuncture can increase the

quality of spermatogenic cells in the testis that has been exposed to heatstroke although it has no effects on

testosterone level.

1 INTRODUCTION

Heatstroke is a high-temperature condition caused

by environment, heat, and direct sunstroke.

Heatstroke can also be called abnormal high

temperature that has a great potential of occurrence

in Indonesia, considering its geographical location

that has a tropical climate. This uncertain heat

condition is exacerbated by the lack of ventilation

and global warming (Ayo et al., 2011).

Infertility problems are often encountered by

adult men and if not treated immediately, will give

an impact on the sterility of reproductive organs.

Recent research results suggest that 8% seek medical

help to treat infertility symptoms (Ashamu et al.,

2010). Infertility disorders are also experienced by

livestock that has economic value in Indonesia,

which has an impact on the decreasing quality and

quantity of livestock sector. One of the most

important causes of infertility besides infectious

diseases is the extreme temperatures caused by

global warming that affect livestock reproduction

system (Boone and Huston, 1963).

Heatstroke can happen in Indonesia.

Climatographic factors in tropical regions make

heatstroke conditions cause infertility cases in

humans and livestock. Temperatures above 27

C can

decrease the quality of spermatozoa and decrease the

rate of conception in livestock (Sari, 2014).

The presence of heatstroke may trigger a

decrease in the immune system of the body and

activate the release of the interleukin-1 cytokine (IL-

1). The release of IL-1 as a compensation of the

immunosuppressed state is followed by increased

body temperature and decreased food intake of each

cell. IL-1 also performs a system of a blockade in

Purnama, M., Rahmaningtyas, I., Putri, A., Lee, A., Tatimma, F. and Masyitoh, H.

Acupuncture Can Increase Spermatogenic Cells in Albino Rats (Rattus norvegicus) Exposed by Heatstroke.

DOI: 10.5220/0007547305750577

In Proceedings of the 2nd International Conference Postgraduate School (ICPS 2018), pages 575-577

ISBN: 978-989-758-348-3

575

the function of gonadal glandular impulses in the

hypothalamus and vice versa stimulate the adrenal

gland axis. The impact of the blockade system on

the gonadal glands decreases the secretion of

Gonadotropin-Releasing Hormone (GnRH), which

also implies the decreased production of Follicle

Stimulating Hormone (FSH) and Luteinizing

Hormone (LH) (Dantzer and Kelly, 2008).

Heatstroke may also increase lipid peroxidase

processes that lead to increased reactive oxygen

species (ROS) and become the beginning of

spermatozoa cell death process (Dobson et al.,

2003).

Acupuncture has been developed as a traditional

medicine technique in overcoming health problems.

This technique can be performed by using

acupuncture needles and laserpuncture. Acupuncture

centers are present at acupuncture meridian points

and specific points associated with a specific disease

(Xie and Preast, 2007).

The Wei-Jian and Ming-Men points are

important in body heat control and when stimulated,

they can control blood flow, reduce heat, and

organize intracellular and extracellular ion channels

(Xie and Preast, 2007). Stimulation at this point can

also protect the body's chi energy from exposure to

heatstroke.

The objective of the study is to prove the

potential of acupuncture to work as an alternative

therapy to reduce body heat and to affect fertility

improvement on quality aspects of spermatogenic

cells and testosterone levels.

2 MATERIALS AND METHODS

2.1 Ethical Clearance

This study was permitted by animal care and use

ethical committee of Faculty of Veterinary Medicine

Universitas Airlangga with registration number

No.686-KE.

2.2 Treatment

In this study, 20 albino rats were used and adapted

one week before treatment. The details of treatment

in experimental animals are as follows: the negative

control group (C-) was not exposed to heatstroke

and acupuncture, positive control group (C+) was

exposed to heatstroke for 20 seconds. Treatment

groups (T1, T2, T3) were exposed to heatstroke and

acupuncture for 5; 10; 20 seconds/rat/day. Incubator

was set to 45

C to conduct a heatstroke effect.

The acupuncture treatment used a specific needle

for the meridian points. The targets of the meridian

points of the reproductive system are Wei-Jian and

Ming-Men points. Acupuncture treatment has been

done along a week.

Albino rats were operated for a minor surgery to

isolate testicle organs. Testicular histopathologic

preparations were prepared in the pathology

laboratory with hematoxylin-eosin staining (HE).

The interpretation was conducted using Johnsen’s

(1970) method of five fields of view in each

replication with 400 magnifications. The scoring

categories are: 1) no germ cells and Sertoli cells; 2)

no germ cells; 3) there are only spermatogonia; 4)

there are only a few spermatocytes; 5) no

spermatozoa and spermatid cells, but many

spermatocytes; 6) few spermatid cells; 7) no

spermatozoa, but many spermatid cells; 8) a few

spermatozoa cells; 9) many spermatozoa but

unorganized spermatogenesis; 10) complete

spermatogenesis and tubular seminiferous.

The blood that has been taken intracardially was

silenced in the test tube to obtain blood serum.

Blood serum was stored in cold temperatures and

prepared for an ELISA test to determine testosterone

levels. Optical value density (OD) and absorbance

results were analyzed.

2.3 Data Analysis

Data from each of the variables in this study were

tested statistically using ANOVA test. When the

results showed significant differences (p<0.05) then

the Duncan test continued.

3 RESULTS AND DISCUSSION

The results of testicular histopathologic readings and

testosterone levels obtained were tabulated

according to the data, then analyzed statistically with

the SPSS v20 for windows program. The mean and

standard deviation (SD) interpretations of testicular

histopathology and testosterone levels are presented

in Table 1.

Table 1: Mean and standard deviation of

spermatogenic cells and testosterone level.

Spermatogenic cells

(x±SD)

Testosterone

level

(x±SD)

C- 8,3

±0,416 1,6

±0,109

C+ 4,2

±0,000 1,4

±0,174

ICPS 2018 - 2nd International Conference Postgraduate School

576

T1 6,2

±0,489 1,4

±0,182

T2 6,6

±0,574 1,6

±0,246

T3 7,6

±0,526 1,6

±0,107

Different superscripts in the same column indicate

significant differences among treatments (p<0.05).

The results showed significantly different

spermatogenic cells (p<0,05) between C+ and C-,

T1, T2, and T3. The treatment groups T1 and T2

were not significantly different (p>0.05).

The results showed no significant difference on

testosterone level (p>0,05) between the treatment

groups. This suggests that there was no effect of

acupuncture therapy on testosterone levels of the

experimental animals exposed to heatstroke.

Acupuncture therapy is expected to improve the

quality of spermatozoa and can be recommended for

treatment since the mass motion repair reaches 70-

80%, progressive individual motion, and semi-dense

concentration. Acupuncture therapy indicates the

improvement and regeneration of germ cells.

Stimulation at the Ming-Men point can restore

the temperature after receiving heatstroke exposure.

Stimulation at the Ming-Men point can open the

orifices of the skin pores, cool the blood, and open

the electrolyte and homeostasis channels. The body's

response to the metabolic process will return to

normal with stimulation at the Ming-Men point,

which is also a relaxation point (Xie and Preast,

2007).

Stimulation at the Wei-Jian point and the Ming-

Men point, especially in the intervertebral region can

improve the performance of the reproductive system.

Ming-Men can restore the function of

spermatogenesis by repairing the damaged germ

cells. The meshing point that also controls the

rehydration setting can cool the reproductive organs

and kidneys. Indication in addition to the return of

reproductive function is also a neutral cell function

in suppressing metabolism after exposure to

heatstroke (Xie and Preast, 2007).

Testosterone hormone is a corticosteroid

derivative hormone, especially in males. Many

testosterone hormones are produced by Leydig cells.

Through the impulse arising from Interstitial Cell

Stimulating Hormone (ICSH) produced by the

anterior pituitary, ICSH will induce Leydig cells to

produce testosterone. Exposure to heatstroke does

not affect the anterior pituitary and suppresses the

secretion of ICSH so that the production of

testosterone by Leydig cells persists (Hafez, 2000).

4 CONCLUSIONS

Based on the results and discussion, it can be

concluded that acupuncture therapy increases the

spermatogenic cells through histopathology of the

testis and has no significant effects on testosterone

levels.

ACKNOWLEDGEMENT

The authors acknowledge The Directorate General

of Higher Education (DIKTI) 2017, The Research,

Technology and Higher Education, the Republic of

Indonesia for providing fund support to carry out

this study.

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