@2024 Afarand., IRAN

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ISSN: 2252-0805    The Horizon of Medical Sciences 2017;23(3):193-199

Background
Over the past decades, androgenic anabolic steroids have become popular among athletes for improving performance and aesthetic reasons. Abuse of androgenic anabolic steroids is not limited to elite athletes, which has been proven to be among amateur athletes as well [1].
Previous Studies
… [2]. Excessive use of androgenic anabolic steroids disrupts the production of testosterone and gonadotropin. In men, suppression of the production of gonadotrophin hormone causes testicular atrophy [3]. In the same line, in the conducted studies, significant reduction in testis weight [4], a significant reduction in testosterone serum levels [5] and severe lesions in the testes [6] have been shown. … [7-11].
Aim(s)
The purpose of this study was to investigate the effect of 6 weeks of resistance training and Boldenon supplements on the expression of 5-alpha reductase and testicular aromatase in Wistar rats.
Research type
This is an experimental research.
Research society, place and time
This study was performed on 30 male Wistar rats aged 12 weeks and mean weight of 59.9 ± 7.94 grams.
Sampling method and number
30 male Wistar rats were obtained from Damghan Scientific Applied Research Institute.
Used devices&materials
The mice were randomly divided into five groups (6 mice in each group) as follows: 1. Control group (without exercise+ placebo injection) 2. Sham group (without exercise + infusion of olive oil) 3. Boldenone supplementation (without exercise+ boldenone injection of 2 mg per kg body weight) 4. Resistance training group (resistance training + placebo injection) 5. Exercise-Boldenone group (resistance exercise + Boldenone injection at 2 mg/kg body weight) Injection of the drug into the animal was carried out with a single insulin syringe once a week, at the appointed time (at 11 am) and one day per week, in the quadriceps and thighs deeply. The control group also received normal saline physiological solution or sodium chloride solution 0.92. Resistance Training Protocol: A resistance training consisted of a 6-week climb of a 76-cm ladder with 47 steps and a width of 19 cm at an angle of 80 degrees with the upper resort (Pars Sports Engineering Call, Girotec, Iran). In order to determine the appropriate weight, the weight of each rats was measured every 4 days. Each session consisted of 3 sets of 5 repetitions that there was a minute rest interval between two sets. The exercise was done after the weight was fastened to the rat. In the first week, the amount of weights fastened to the tail of rats was 50% of a maximal repeat (1RM) of each animal that was calculated the day before the start of the resistance training. This amount was increased 10% per week that it reached to 100% in the final week. Animals became familiar with climbing the ladder during the 2 weeks prior to the start of the training that they were forced manually to climb in the case of refusal. The final load was considered as the maximum carrying capacity of that meeting [12]. Tissue sampling and enzyme gene expression measurement: After anesthesia, an autopsy was performed by fixing the animal on the rodent surgery board, and immediately the testicular tissue was removed. Sampling from testicular tissue from 5 groups were performed after intervention of independent variables and changes in gene expression of enzymes in the testicular tissue were studied and then compared. Full RNA extraction: for full RNA extraction, first, the tissue was extracted from the temperature of -80 C and after weighing in phosphate buffer (0.1% phemylmethylsulfonyl fluoride, PMFS), it was well homogenized by homogenizer and isolated with a isolating kit with a catalogue number 11667157001(Roche, Germany), and according to its instructions, all tissue RNAs were extracted. Then, the extracted RNA was solved in 50 micro liter of RNase-free water and the RNA quality purified by spectrophotometer was evaluated. Also, the RNA extracted in 8.0 % agarose gel was electrophoresed and stained with ethidium bromide to confirm and verify the quality and quantity of RNA, and after assuring the correctness of the purified RNA was entered into the next stage. Production of complementary DNA (cDNA): One microgram of extracted RNA for producing cDNA was used by AccuPowerR CycleScriptRT PreMix (dN6) kit with catalog No. 2044 (Bioneer, South Korea). In order to produce cDNA, master mix including RNase-free water and random primer (50 mM) were used. In accordance with the kit's instructions, the mixture was first incubated at 45 ° C for 60 minutes and then incubated for 5 minutes at 95 ° C (Table 1). Following the preparation of cDNA as a template sample using PCR real time method and the machine model 7500 (ABI; USA), the changes of gene expression (2^-ΔΔCt) in 5α-reductase and aromatase enzymes were measured in accordance with the machine's instructions. Statistical analysis: The Kolmogrov-Smirnov test was used to assure the normal distribution of data. Then, t-test was used to examine the intra-group changes and one-way ANOVA and Tukey's post hoc test were used for investigating intergroup changes. All statistical operations of the research was performed using SPSS 22 software.
Findings by Text
The mean weight of rats in all groups was significantly increased during the post-intervention period compared to pre-intervention period (p=0.0001). However, there was no significant difference between the mean weights of rats in the groups after intervention (p> 0.05, Table 2). After intervention, there was a significant difference between the mean of 5α-α-reductase gene expression of male Wistar rats in different groups (p = 0.0001). The changes of 5α-α-reductase gene expression in the Boldenone group (p = 0.046) and exercise-Boldenone group (p = 0.001) compared to the control group, and in the training-boldenone group compared to exercise-resistance group (p=0.019) were significant. However, there was no significant difference in the expression of this enzyme in the exercise-boldonone group and in the resistance-exercise group compared to the boldonone group (p>0.05; Figure 1). Also, there was a significant difference between the mean expression of the aromatase enzyme gene of male Wistar rats in different groups of research (p=0.0001). Changes in aromatase gene expression in the resistance training group (p=0.017) and boldorone training (p=0.0001) were significantly higher than the control group. However, there was no significant difference in the expression of aromatase gene in the resistance training group and the boldenon exercises compared to the boldonone group and in the boldonone group compared to the resistance training group (p>0.05).
Main comparison to the similar studies
… [12-25]. In many strength athletes, after the use of anabolic steroids, body weight gain has often been reported. Most studies show that body weight may increase as a result of short-term use (less than 10 weeks) of androgenic anabolic steroids [26, 27]. The highest body weight gain was reported by Casner et al. after 6 weeks of administration of stanozolol [28]. However, in a case report, there has been a significant increase over the course of two-year period of administration of androgenic anabolic steroids [29]. This is probably due to the structural process of body tissues by the boldenone supplementation as a result of an increase in muscle size that results from a positive nitrogen balance by stimulating protein synthesis and reducing protein degradation [30]. Also, an increase in body weight may be attributed to an increase in total protein serum and globulin protein which indicates improvement in health and immunity of body. However, in the present study, the total protein and globulin concentration levels have not been measured.
Suggestions
It is suggested that in the same study, the effect of 5α-reductase and aromatase inhibitors and stimulants be investigated simultaneously. Also, levels of sex hormones are measured along with 5-alpha-reductase and aromatase.
Limitations
Lacking control over the exact diet (measuring energy intake and consumption) is one of the limitations of this research.
Conclusions
Boldenone supplements plus 6 weeks of resistance training can increase levels of expression of 5α alpha reductase and aromatase genes in testicular tissue in Wistar rats.
Acknowledgments
The authors express their gratitude to the Deputy of Research of the Azad University of Ayatolloh Amoli Branch.
Conflict of interest
Non-declared
Ethical Permissions
This research was carried out with the approval of the Ethics Committee at the Islamic Azad University of Ayatollah Amoli Branch.
Funding Sources
This research is based on a graduate dissertation with an identification code of 23921404941004 and supported by the Deputy of Research of the Islamic Azad University of Ayatollah Amoli Branch.

TABLES and CHARTS

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