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ORIGINAL ARTICLE
Year : 2018  |  Volume : 6  |  Issue : 2  |  Page : 51-58

Effects of mobile phone radiation and exercise on testicular function in male Wistar rats


Department of Physical Activity and Health Promotion, Faculty of Medicine, University of Rome, Tor Vergata,Rome, Italy

Date of Web Publication25-Feb-2019

Correspondence Address:
Mr. Chidiebere Emmanuel Okechukwu
Department of Physical Activity and Health Promotion, Facoltà Di Medicina, Università Degli Studi Di Roma Tor Vergata, Rome
Italy
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Source of Support: None, Conflict of Interest: None


DOI: 10.4103/njecp.njecp_18_18

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  Abstract 


Background: The aim of this study was to investigate the effects of mobile phone radiation and exercise on testicular function in male Wistar rats. Methods: Twenty male Wistar rats weighing 150–198 g were used for this experiment. The animals were divided into four groups (n = 5). The first group was the control group, the second group was exposed to mobile phone radiation for 6 h daily, the third group was subjected to swimming >three times a week and >30 min each session or >90 min a week, and the fourth group containing five rats was exposed to mobile phone radiation for 6 h daily and was subjected to swimming for >three times a week and >30 min each session or >90 min a week. This experiment lasted for 30 days. A Nokia 1280 cell phone was used to emit electromagnetic waves, and the cages of Groups 2 and 4 were surrounded by aluminum foils to focus and limit the waves to the interior of the cages, with the phone being placed 0.5 cm under the cage. A radiofrequency radiometer was kept close to the cages to detect the cell phone radiation. Results: Short-term exposure of male Wistar rats to mobile phone radiation (6 h/day × 30 days at 1.6 W/kg specific absorption rate) led to a statistically insignificant (P > 0.05) decrease in the serum testosterone levels and testicular weight, whereas exercise (> three times a week and >30 min each session or > 90 min a week) in male Wistar rats led to statistically nonsignificant (P > 0.05) increase in the testosterone levels and testicular weight. Conclusions: The findings of this study indicated that short-term exposure of mobile phone radiation leads to a statistically nonsignificant decrease in serum testosterone levels and testicular weight, whereas regular exercise leads to a statistically nonsignificant increase in the testosterone levels and testicular weight.

Keywords: Exercise, male Wistar rats, mobile phone radiation, swimming, testicular function, testicular weight, testosterone levels


How to cite this article:
Okechukwu CE. Effects of mobile phone radiation and exercise on testicular function in male Wistar rats. Niger J Exp Clin Biosci 2018;6:51-8

How to cite this URL:
Okechukwu CE. Effects of mobile phone radiation and exercise on testicular function in male Wistar rats. Niger J Exp Clin Biosci [serial online] 2018 [cited 2019 Sep 22];6:51-8. Available from: http://www.njecbonline.org/text.asp?2018/6/2/51/252843




  Introduction Top


Mobile phones are radio devices that transmit and receive radiofrequency radiation at 900–2000 MHz. Electromagnetic waves (EMWs) emitted by mobile phones can damage sex organs and cause hormonal disorders. Cell phone technology is an integral part of day-to-day life, and its use is not only restricted to voice conversations but also conveying news, high-resolution pictures, and the Internet.

Exercise consists of activities that are planned and structured that maintain or improve one or more of the components of physical fitness. Dynamic exercise is defined as skeletal muscle contractions at changing lengths with rhythmic episodes of relaxation. The magnitude of testosterone release during exercise can be varied by exercise mode, intensity, and duration.[1] An acute bout of exercise rapidly elevates testosterone level in circulation. This response is mediated by increased sympathetic activity during the exercise and lactate, a metabolite produced from exercised skeletal muscle during anaerobic glycolysis.[2]

No study has been carried out before to determine the effects of mobile phone radiation and exercise on serum testosterone levels, testicular weight, and testicular architecture in male Wistar rats as a combined research work. Nigeria has the highest number of mobile phone users in Africa, with more than 150 million subscribers, and recent studies have shown that electromagnetic radiation emitted by mobile phones has an adverse effect on male fertility.

Lack of exercise and obesity adds to the general effect of aging and low testosterone levels.[3] Recent studies have shown that testosterone is more responsive to higher intensity exercise[4] and a longer duration of exercise.[5],[6],[7],[8],[9] The aim of this study was to investigate the effects of mobile phone radiation and exercise on testicular function in male Wistar rats.

This study gives an insight into the possible effects of mobile phone radiation and exercise on serum testosterone levels, testicular weight, and testicular architecture in male Wistar rats, which can be linked to adult human males that make use of mobile phones frequently in their homes and offices, for example, bankers, telecommunication workers, customer care servicemen, doctors, pilots, receptionists, and male individuals, who are fond of putting their mobile phones on their side pockets which is very close to their testes. This study will also help researchers to understand testosterone response to exercise with regard to the athletes and individuals who exercise regularly. Further studies will determine if mobile phone radiation and exercise has a significant effect on testicular function.


  Methods Top


Study design

This experimental study was conducted at the Department of Human Physiology Laboratory, Faculty of Basic Medical Sciences, Nnamdi Azikiwe University, Anambra State, Nigeria, with a total of 20 male Wistar rats that were allowed to acclimatize to the environment for 1 week; they were subjected to exercise for 1 month and exposed to mobile phone radiation for 30 days. The rats were fed ad libitum with standard rat diet and distilled water. The animal room was well ventilated within a temperature range of 25°C–27°C. The rats were labeled and divided into four groups. Their weight was recorded weekly before and during experimentation.

Study procedure

Twenty male Wistar rats weighing 150–198 g were divided into four groups (n = 5). The first group was the control group, the second group was exposed to mobile phone radiation (900–1800 MHz) for 6 h daily, the third group containing five rats was subjected to swimming > three times a week and >30 min each session or >90 min a week, and the fourth group was exposed to mobile phone radiation (900–1800 MHz) for 6 h daily and was subjected to swimming >three times a week and >30 min each session or >90 min a week; this experiment lasted for 30 days. A Nokia 1280 cell phone (Nokia corp. Announced in November 2009. Released in March 2010. Shenzhen, China) was used to emit EMW radiation, and the cage of Groups 2 and 4 was surrounded by aluminum foils to focus waves and limit the electromagnetic field to the interior of the cages. During wave exposure, the cell phone (placed 0.5 cm under the cage) was set in different modes including call, missed call, and turned-on mode (without real talk).

Measurement of mobile phone radiation

A dual-band (900 MHz, 1800 MHz) Nokia 1280 mobile in receiving mode was used as the radiation source in this experiment. Its specific absorption rate (SAR) is 1.15 based on 1.6 W/kg averaged over 1 g of body tissue or 0.81 based on 2.0 W/kg averaged over 10 g of body tissue. The lower the SAR of a phone, the better it is. Therefore, using a phone with SAR of 1.15 represents a near worst-case scenario. Nokia Asha 202 mobile, also of the same dual band, with SAR 1.01 based on 1.6 W/kg averaged over 1 g of body tissue or 1.11 based on 2.0 W/kg averaged over 10 g of the body tissue, was used as the transmitting phone in this experiment. The SAR of both the receiving and transmitting mobile phones fall within the International Standard of exposure limit for public exposure which is 1.6 W/kg radiation rating averaged over 1 g of the body tissue. A radiofrequency radiometer was kept close to the cages to detect the cell phone radiation.

Animal sacrifice

At the end of the 30 days' experiment, the animals were sacrificed between 9:00 am and 11:00 am to minimize the diurnal fluctuations of hormonal secretion. The animals were anesthetized using chloroform and sacrificed by cervical dislocation 12 h after the last experiment, and their testes were excised following abdominal incision and were weighed with an electronic weighing machine, fixed in 10% formalin for histological analysis.

Serum testosterone analysis

Blood samples were collected through cardiac puncture, stored in plain tubes without anticoagulants, and allowed to clot. The clotted blood samples were centrifuged at 2000 rpm for 15 min to obtain serum. The serum was stored at −20°C until the analysis. Serum testosterone level was measured using enzyme-linked immunosorbent assay (ELISA, AccuBind ELISA Microwells Testosterone test system with the product code: 3725-300) kits manufactured by Monobind Inc., Lake Forest, CA, USA. The sensitivity of hormone detected per assay tube was 0.05 ng/ml.

Histological analysis

The rats' testes were carefully dissected out following abdominal incision and fixed in 10% formalin. The left testes of the rats in all the groups were processed routinely for paraffin embedding. 5-μ sections were obtained with rotatory microtome. The tissue samples were embedded in paraffin and their 5-mm cross sections were stained with hematoxylin and eosin. All the slides were examined under a light microscope (×400) and sections were observed.

Measurement of testicular weight

The left and right testes of the rats in all the groups were dissected out, freed from adherent tissues, and weighed up to the nearest 0.001 g on a Mettler analytical balance (PE 1600, Mettler Instrument AG, Switzerland). The result was computed per 100 g of the body weight.

Statistical analysis

The data collected during the experiment were analyzed by one-way ANOVA test determined at P < 0.05 using SPSS software package version 20 (IBM Corp. Released in 2011. IBM SPSS Statistics for Windows, Version 20.0, Armonk, NY, USA). Dependent paired t-test was used to compare the results within the groups. All the results were presented as mean and standard error of the mean (SEM), n = 5.


  Results Top


Effect of mobile phone radiation and exercise on serum testosterone levels of male Wistar rats

[Table 1] shows ANOVA analysis comparing the serum level of testosterone based on groups' treatment. When the mean of the serum testosterone concentration of Group 1 (control group) (3.18 ± 0.41) was compared to that of Group 2 (1.74 ± 0.73) and Group 3 (3.44 ± 0.49) which were exposed to mobile phone radiation only for 30 days and exercise only for 1 month, respectively, there was no statistically significant difference (P > 0.05). However, when the mean of serum testosterone concentration of Group 1 (control group) (3.18 ± 0.41) was compared to that of Group 4 (2.84 ± 0.33) which was exposed to mobile phone radiation and exercise for 30 days, there was a slight statistically significant difference (P > 0.05) [Figure 1].
Table 1: ANOVA analyses comparing the serum level of testosterone based on group treatment

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Figure 1: Comparison of serum testosterone based on group treatment

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Effect of mobile phone radiation and exercise on testicular weight of male Wistar rats

[Table 2] shows the ANOVA analysis comparing the left testicular weight and right testicular weight of the various groups based on groups' treatment. When the mean of left testicular weight of Group 1 (control group) (1.27 ± 0.04) was compared to that of Group 2 (1.11 ± 0.04), which was exposed to mobile phone radiation only for 30 days, and to that ofGroup 3 (1.37 ± 0.03), which was subjected to exercise only for 1 month, there was no statistically significant difference (P > 0.05) [Figure 2]. However, when the mean of left testicular weight of Group 1 (control group) (1.27 ± 0.04) was compared to that of Group 4 (1.53 ± 0.07) which was exposed to mobile phone radiation and subjected to exercise for 1 month, there was an increased statistically significant difference (P < 0.05). For the right testicular weight, when the mean of right testicular weight of Group 1 (control group) (1.27 ± 0.03) was compared to that of Group 2 (1.13 ± 0.05), which was exposed to mobile phone radiation only for 30 days, and to that of Group 3 (1.38 ± 0.06), which was subjected to exercise only for 1 month, there was no statistically significant difference (P > 0.05). However, when the mean of right testicular weight of Group 1 (control group) (1.27 ± 0.03) was compared to the mean of Group 4 (1.50 ± 0.06) which was exposed to mobile phone radiation and subjected exercise for 1 month, there was an increased statistically significant difference (P < 0.05).
Table 2: ANOVA analysis comparing testicular weight/100 g of body weight based on group treatment

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Figure 2: Comparison of the left and right testicular weights based on group treatment

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Histological examination of testes

The testes of the rats in the control Group 1 [Figure 3] contain a good number of seminiferous tubules with connecting tissue separating them, boundary tissue consisting of the outer layer of collagen fiber, normal-sized seminiferous tubules which were full of spermatogenic cells with scanty interstitial tissue, and few Leydig cells and intact germinal epithelial layers of adjacent seminiferous tubules.
Figure 3: Testicular section of Wistar rat testis (control group) not exposed to mobile phone radiation and was not subjected to exercise (H and E, ×400) shows normal spermatogenesis ( first and second red arrow from the left) within seminiferous tubule. Intact germinal epithelial layers of adjacent seminiferous tubules (arrow on the right side), Leydig cells at the interstitial spaces,  Sertoli cells More Details, spermatogonia, spermatocytes, spermatids, and numerous mature spermatozoa are clearly visible

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Light microscopic examination of the testes sections of rats in Group 2 [Figure 4] exposed to mobile phone radiation for 6 h per day for 4 weeks showed alterations in the seminiferous tubules of testis compared with the control group. Inhibition of spermatogenesis was observed. The number of sperms was reduced; some tubules showed spermatocytes. The Leydig cells were degenerated, and the intertubular blood vessels were dilated and congested. Most of the tubules were devoid of sperms, and in others, the sperms were scattered randomly in the tubules. Although other phases of the cycle could be observed, some tubules present signs of necrosis.
Figure 4: Testicular section of Wistar rat testis exposed to mobile phone radiation (H and E, ×400) shows features of incomplete spermatid maturation arrest (mature spermatozoa are almost totally absent) and shows the seminiferous tubule, the Leydig cells at the interstitial spaces are small and few in number, Sertoli cells, spermatogonia (curved arrow), spermatocytes (straight arrow), and spermatids (arrowhead). Mature, viable spermatozoa are almost completely absent

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Light microscopic examination of the testes section of rats in Group 3 [Figure 5] subjected to exercise for 1 month showed the full spermatogenic activity of the seminiferous epithelium cycle represented by spermatogonia, primary spermatocytes, secondary spermatocytes, and spermatids as well as nutritive Sertoli cells. Seminiferous tubules were separated by intertubular connective tissues with Leydig cells. The lumen was filled with spermatozoa.
Figure 5: Testicular section of male Wistar rat testis subjected to exercise (H and E, ×400) shows normal active spermatogenesis, there are Sertoli cells, spermatogonia (curved arrow), and many Leydig cells at the interstitial spaces, spermatocytes, spermatids (straight arrow), and numerous mature spermatozoa (arrowhead)

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Light microscopic examination of the testes section of rats in Group 4 [Figure 6] exposed to mobile phone radiation and exercise for 1 month revealed some alterations in both the interstitial tissue and seminiferous tubules. The detachment between the adjacent seminiferous tubules was observed at several places, and the number of Leydig cells at interstitial space was decreased.
Figure 6: Testicular section of Wistar rat testis exposed to mobile phone radiation and exercise (H and E, ×400) shows a seminiferous tubule with low spermatogenesis. The number of Leydig cells at the interstitial spaces are slightly decreased. Sertoli cells, spermatogonia (curved arrow), spermatocytes, and spermatids (straight arrow). However, there is a significant reduction in the number of mature spermatozoa (arrowhead)

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The overall research findings of this study [Figure 7] prove that exercise (>3 times a week and >30 min each session or >90 min a week) leads to a statistically nonsignificant increase in testicular weight, an increase in Leydig cells, and an increase in spermatogenesis and serum testosterone levels, whereas mobile phone radiation (6 h/day × 30 days at 1.6 W/kg SAR) leads to a statistically nonsignificant decrease in testicular weight, decrease in Leydig cells, and decrease in spermatogenesis and serum testosterone levels. The graph of mean testosterone (ng/dl) of the groups [Figure 8] shows an increase in serum testosterone levels in the group that performed exercise when compared to the control group, whereas there was a decrease in the serum testosterone levels in the group exposed to mobile phone radiation when compared to the control group. There was a slight decrease in the serum testosterone levels of the group exposed to both mobile phone radiation and exercise when compared to the control group. These findings prove that exercise leads to an increase in male fertility, whereas mobile phone radiation leads to a decrease in male fertility.
Figure 7: Schematic summary of the research findings

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Figure 8: Mean of testosterone levels of the groups

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  Discussion Top


According to the results of this study, exposure of male Wistar rats to mobile phone radiation 6 h daily for 30 days leads to a statistical decrease in the serum testosterone levels, which was nonsignificant (P > 0.05) when compared to the control group, but it might be significant if the duration of the experiment was extended beyond 30 days. This is nearly similar to the reports of Oyewopo et al., 2017,[10] who stated that mobile phone radiation causes a significant decrease in serum testosterone concentration. There was a statistical increase in the serum testosterone levels in male rats subjected to exercise only, but it was statistically nonsignificant (P > 0.05) when compared to the control group. However, it might be significant if the duration of the experiment was extended beyond 1 month; this is contrary to the findings of Hu et al., 1999,[11] who observed a significant reduction in testosterone levels in rats subjected to continuous swimming for 3 weeks. Testosterone levels were restored to normal following 6 weeks of training, suggesting an adjustment to training on luteinizing hormone secretion in the endocrine system that was associated with a negative feedback mechanism. However, there was a slight statistical decrease in serum testosterone levels in male Wistar rats exposed to mobile phone radiation and subjected to exercise when compared to the control group, but it was statistically nonsignificant (P > 0.05).

Short-term exposure of male Wistar rats to electromagnetic field (EMF) radiation emitted from a Nokia 1280 cell phone for 30 days caused a nonsignificant (P > 0.05) decrease in testicular weight. Similarly, Aminollah et al., 2015,[12] assessed the effects of EMF with an intensity of 1 mT, 50 Hz on reproductive variables and sex hormones in male rats and found a nonsignificant decrease in testicular weight. Swimming for greater than three times a week and >30 min each session or >90 min a week led to a statistically nonsignificant increase (P > 0.05) in the testicular weight of male Wistar rats subjected to exercise when compared to the control group. However, there was a statistically significant increase (P < 0.05) in testicular weight of male rats exposed to mobile phone radiation and subjected to exercise when compared to the control group.

Microscopic changes such as reduction in the mean seminiferous tubule diameter, vacuolar degeneration and desquamation of the seminiferous epithelium, maturation arrest in the spermatogenesis of the peripheral tubules, and reduction in the number of Sertoli and Leydig cells were observed in the testis of the male rats exposed to mobile phone radiation, and this is similar to the findings of Çetkin et al. 2017,[13] Oh et al. 2018,[14] Forgács et al. 2006,[15] Kesari and Behari 2010,[16] Kesari and Behari 2012,[17] Kesari et al. 2011,[18] Kumar et al. 2012,[19] Meo et al. 2010,[20] Ozguner et al. 2005,[21] and Salama et al. 2009.[22]

Microscopic changes such as full spermatogenic activity of the seminiferous epithelium cycle represented by spermatogonia, primary spermatocytes, secondary spermatocytes, and spermatids as well as nutritive Sertoli cells were observed in the testis of rats subjected to exercise. However, microscopic changes in the testis of rats in the test group exposed to mobile phone radiation and subjected to exercise showed alterations in both the interstitial tissue and seminiferous tubules. The detachment between the adjacent seminiferous tubules was observed at several places, and the number of Leydig cells at interstitial space was observed as decreased.

Limitations

The present study has limitations. The sample size was relatively small, and the study was performed at a single institution. The duration of the study was less; therefore, there is a need for further studies and long-term research on the effects of mobile phone radiation and exercise on testicular functions, which may likely have a significant effect on the testicular function.

Future directions and recommendations

Based on this animal study, the excessive use of mobile phone over a long period of time should be avoided because such effects are also expected in humans. Mobile phones should not be kept inside the pocket for a long period of time due to its thermal effect and proximity to the testes. The use of hand-free materials and gadgets limits direct contact of cell phones to the body, for example, the Bluetooth earpiece. Shield case or pulse should be encouraged by the manufacturers. The cell phone should be used only where the reception is good. The weaker the reception, the more the power the phone must use to transmit and the more power it uses, the more radiation it emits, and the deeper the dangerous radio waves penetrate the body. Ideally, a user should only use the phone with full bars and good reception. The damage from cell phone exposure will take many years to surface, and there are rarely any initial symptoms, just like cancer. Service providers and mobile phone manufacturers should provide research funds to tertiary and biomedical/biophysics research institutes worldwide to conduct researches related to the effects of radiofrequency radiation emitted by mobile phones in humans as a result of the increased global use of mobile phones.

At least 2 h and 30 min of moderate aerobic activity per week should be recommended as a therapy for men with low testosterone levels, aging males, and men whose occupation is sedentary. Swimming at least three times a week over a long period of time can lead to an increase in male fertility.


  Conclusions Top


The findings of this study indicated that short-term exposure of mobile phone radiation (6 h/day × 30 days at 1.6 W/kg SAR) leads to a statistically nonsignificant decrease in serum testosterone levels and testicular weight. Microscopic examination of the rat testis exposed to mobile phone radiation showed alterations in the seminiferous tubules of the testis, inhibition of spermatogenesis, degeneration of Leydig cells, and dilation and congestion of intertubular blood vessels. Exercise (>three times a week and >30 min each session or >90 min a week) leads to a statistically nonsignificant increase in the testosterone levels and testicular weight. Microscopic examination of the rat testis subjected to exercise showed the full spermatogenic activity of the seminiferous epithelium cycle represented by spermatogonia, primary spermatocytes, secondary spermatocytes, and spermatids as well as nutritive Sertoli cells was observed.

Acknowledgment

Special thanks to Prof. Ed Nwobodo, Dr. Vincent Igbokwe, the academic and nonacademic staff of the Department of Human Physiology, Nnamdi Azikiwe University, for their contributions toward this project.

Financial support and sponsorship

This study was funded by Mr. Okechukwu Chidiebere Emmanuel who picked interest in the study.

Conflicts of interest

There are no conflicts of interest.



 
  References Top

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    Figures

  [Figure 1], [Figure 2], [Figure 3], [Figure 4], [Figure 5], [Figure 6], [Figure 7], [Figure 8]
 
 
    Tables

  [Table 1], [Table 2]



 

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