Research Article - Biomedical Research (2017) Volume 28, Issue 8
Effects of single dose administered nandrolone decanoate on serum cytokine levels and some biochemical parameters in male and female ratsErdal Tasgin1*, Sefa Lök1, Seyfullah Haliloğlu3, Nagehan Demir2 and Hale Ergin3
3Department of Biochemistry, Veterinary Faculty, Selcuk University, 42075 Konya, Turkey
- *Corresponding Author:
- Erdal Tasgin
School of Physical Education and Sport, Turkey
Accepted date: January 24, 2017
The aim this this study was to determine that administration of single higher dose nandrolone decanoate (40 mg/kg) on serum IL-1β, TNF-α, IL-10 and IL-6 levels and some biochemical parameters during 24 h in male and female rats. The blood samples were taken from 5 male and female rats to obtain their starting values. Then the blood samples were taken from 6 rats in sampling time at 1st, 2nd, 4th, 8th, 12th and 24th h following the nandrolone decanoate administration. There were significant increases determined in the serum IL-1β levels, numerical increases in the TNF-α levels whereas decreased IL-6 levels were determined. IL-10 levels did not change during experimental period. While serum GGT and BUN levels increased towards the last h of the study, increased ALP levels were observed only female rats. There was no any significance changes were determined in the other biochemical (ALT, AST, CKMB and LDH) values. As a result, increased IL-1β and TNF-α levels, proinflammatory cytokines, and some biochemical parameters may be reflect that long term used nandrolone decanoate causes organ damages and effect immune system function. However, effects of long term used nandrolone decanoate on the immune system and organ functions should be detailed evaluated.
Nandrolone, Cytokines, Organ damages
It is stated that use of doping has increased significantly in recent years, and it harms human health and at the same time, it also affects ethical values of sport negatively . Doping refers to taking any impurity into body illegally by athletes in order to gain an advantage over competitor in contests and competitions . After the competitions, a science committee follows traces of various performance enhancers with blood and urine tests. This is very important in terms of protecting justice of the sport with athletes . The fight against doping has become a political issue around the world for more than 60 years .
While AAS (Anabolic androgenic steroids) which are ones of synthetic derivatives of male sex hormones and testosterone [5-8] had being used for burns and traumas, radiation therapy and chronic weight loss diseases since 1940s, They became even more important in treatments of AIDS related burnout syndrome and kidney diseases after 1985 .
Materials and Methods
Totally 35 male and 35 female rats were used. 5 male and female rats were accepted as control time (0 hour), and rested 30 male and female rats were received at the doze of 40 mg/kg nandrolone decanoate (IP). After treatments, blood samples were collected at 1, 2, 4, 8, 12 and 24 hour from cardiac puncture under anesthesia. Serum ALP, ALT, AST, CK-MB, GGT, LDH and BUN levels were measured with auto-analyzer, while serum TNF-α, IL-1β, IL-6 and IL-10 levels was determined with ELISA reader. Data were evaluated by ANOVA and Duncan test. P<0.05 was accepted as statistically significant.
Serum biochemical values of male and female rat are shown in Tables 1 and 2, respectively. Increased GGT and BUN levels were determined (P<0.05) in male rats, whereas statistically significant fluctuations were determined (P<0.05) in ALP, GGT, BUN and LDH levels in female rats.
|Parameters||0 h||1 h||2 h||4 h||8 h||12 h||24 h|
|ALP, U/L||120.6 ± 16.0||96.0 ± 10.6||138,5 ± 17.5||110,7 ± 14.5||113,4 ± 19.7||91.8 ± 19.3||116.7 ± 15,74|
|ALT, U/L||57.3 ± 5.71||52.0 ± 4.39||57.0 ± 4.14||63.8 ± 8.73||51.1 ± 6.95||48.0 ± 3.98||54.45 ± 6.46|
|AST, U/L||95.1 ± 16.5||100 ± 14.5||120 ± 19.7||154 ± 26.1||104 ± 13.30||94.0 ± 11.9||112.3 ± 14.4|
|CKMB, U/L||743 ± 160||806 ± 191||730 ± 263||1421 ± 474||674 ± 217||779 ± 251||952 ± 234|
|GGT, U/L||1.06 ± 0.16b||1.83 ± 0.40ab||2.33 ± 0.80ab||2.00 ± 0.36ab||1.50 ± 0.22ab||1.16 ± 0.16b||2.84 ± 0.66a|
|LDH, U/L||578 ± 87.7||485 ± 85.5||505 ± 164||731 ± 180||387 ± 55.2||461 ± 86.8||554 ± 148|
|BUN mg/dl||48.0 ± 1.54c||59.0 ± 3.93abc||54.3 ± 3.33abc||62.6 ± 3.45ab||51.1 ± 2.02bc||66.1 ± 2.46a||64.4 ± 4.6ab|
Table 1. Serum biochemical parameters of male rats after Nandrolon decanoate treatment (mean ± SE).
|Parameters||0 h||1 h||2 h||4 h||8 h||12 h||24 h|
|ALP, U/L||74.5 ± 2.98ab||68.6 ± 4.75b||69.3 ± 4.87ab||62.5 ± 11.4b||65.0 ± 8.98b||58.3 ± 4.91b||103 ± 11.9a|
|ALT, U/L||98.0 ± 28.6||126 ± 23.5||88.0 ± 21.0||72.6 ± 14.7||66.1 ± 5.81||79.0 ± 10.2||76.6 ± 15.0|
|AST, U/L||152 ± 24.4||205 ± 44.2||159 ± 28.3||122 ± 30.2||117 ± 15.5||154 ± 18.6||165 ± 30.1|
|CKMB,U/L||814 ± 187||1223 ± 575||843 ± 132||538 ± 49||778 ± 228||707 ± 198||1262 ± 580|
|GGT, U/L||2.16 ± 0.47ab||1.50 ± 0.22ab||1.16 ± 0.47b||1.66 ± 0.33ab||1.24 ± 0.22b||1.33 ± 0.21b||3.66 ± 0.98a|
|LDH, U/L||891 ± 101a||774 ± 154ab||523 ± 74ab||287 ± 43b||458 ± 142ab||573 ± 138ab||783 ± 182ab|
|BUN mg/dl||59.8 ± 12.7ab||48.6 ± 2.26b||59.3 ± 2.33ab||55.0 ± 3.84ab||54.6 ± 1.68ab||78.6 ± 4.33a||53.8 ± 1.88b|
Table 2. Serum biochemical parameters of female rats after Nandrolon decanoate treatment (mean ± SE).
Serum cytokine levels are presented in Tables 3 and 4, respectively. Increased IL-1β levels are determined (P<0.05) in male rats, whereas statistically significant fluctuations were determined (P<0.05) in IL-6 levels. Increased IL-1β and decreased IL-6 levels were measured (P<0.05) in female rats.
|Parameters||0 h||1 h||2 h||4 h||8 h||12 h||24 h|
|TNF-α||4.27 ± 0.76||4.54 ± 1.20||4.74 ± 1.34||4.40 ± 0.96||5.41 ± 1.74||6.08 ± 1.86||5.76 ± 1.44|
|IL-1β||7.52 ± 1.94b||7.24 ± 1.82b||9.68 ± 1.56a||8.74 ± 1.46ab||10.12 ± 2.94a||9.85 ± 1.44a||9.72 ± 1.52a|
|IL-6||24.48 ± 2.64a||24.90 ± 3.57a||20.48 ± 2.54a||17.1 ± 3.45ab||22.34 ± 5.60a||14.84 ± 3.86b||18.42 ± 2.78ab|
|IL-10||6.84 ± 1.08||6.76 ± 1.14||7.80 ± 1.06||8.08 ± 2.14||7.96 ± 3.08||8.48 ± 2.17||8.12 ± 2.64|
Table 3. Serum cytokine levels of male rats after Nandrolon decanoate treatment (mean ± SE).
|Parameters||0 h||1 h||2 h||4 h||8 h||12 h||24 h|
|TNF-α pg/mL||4.86 ± 0.42||4.78 ± 1.64||5.15 ± 1.06||5.60 ± 0.96||4.88 ± 1.47||5.90 ± 1.64||5.94 ± 1.86|
|IL-1β pg/mL||6.46 ± 1.25b||6.64 ± 1.48b||8.88 ± 1.94a||8.12 ± 2.15a||6.88 ± 1.06b||8.19 ± 1.22a||8.72 ± 2.92a|
|IL-6 pg/mL||20.12 ± 3.85a||19.46 ± 2.98a||18.42 ± 2.08a||16.54 ± 1.13a||17.55 ± 3.72a||16.57 ± 2.92ab||14.18 ± 2.44b|
|IL-10 pg/mL||6.78 ± 1.20||6.66 ± 2.16||7.84 ± 2.26||6.98 ± 2.14||7.89 ± 2.85||7.92 ± 3.02||8.06 ± 2.54|
Table 4. Serum cytokine levels of female rats after Nandrolon decanoate treatment (mean ± SE).
Today, sudden cardiac death due to cardiovascular disease takes an important place in causes of death . Because use of anti-androgenic drugs for treatment in medicine have increased rates of death due to cardiovascular function [13,14], this situation has revealed that there is a need for more in-depth investigation of the effects of androgenic hormones. It has been seen that application of testosterone above physiological doses decreases eNOS activity and increases oxidative stress . As a matter of fact, in relation to the research done, it is reported that rate of death due to cardiovascular diseases has increased among ones misusing androgens and the reason of this might be due to endothelial dysfunction occurring .
Although it is reported that nandrolone decanoate increases TNFα, IL-1β and IL-4 production in cell cultures and suppresses IL-6 mRNA expression [17-19], in some studies, it has been determined that it has no effect on TNFα and IL-1β mRNA expression and IL-2, IL-3 and IL-10 productions [18,20,21]. On the other hand, it has been reported that AAS inhibit activation of nuclear factor Kappa B (NF-κB)  allowing expression of genes encoding proinflammatory cytokines .
The study done has shown that there is no statistical difference related to serum cytokine levels between male and female rats during first 24 h. After administration of ND, it has been determined in both genders that IL1-β levels increased after 2 h following administration (p<0.05) and non-statistical numerical increase in TNF-α levels was observed. It has been seen that results obtained are compatible with findings of some researchers [17-19], they are not compatible with findings of Corrales et al. stating that there is a decrease in IL1-β and TNF-α levels in individuals receiving testosterone therapy. It has been interpreted that ND has much more androgenic effects than testosterone and this is the reason of using high dose of ND in the study and also 24-hour variation might vary. Also, Parrillo et al. and Dinarello have reported that IL1-β and TNF- α levels might be associated with multiple organ failure. In the light of this information, it has been thought that these increases observed in these two cytokines might be sign of results that may cause multiple organ failures.
In the study done, decreases in serum IL-6 levels have been observed in male and female rats during the h following ND administration and it has been observed that this decrease is compatible with some studies [17-19]. There has not been any change observed in IL-10 levels in male and female rats as compatible with findings of Thompson et al. and when results have been analyzed, it has been seen that although it doesn't make statistical sense, there is numerical increase as compatible with statements of Corrales et al.
In many studies in which effects of ASS on liver enzymes were researched, enzyme activities were examined and different results were obtained in terms of these activities [20-26]. In some studies, any change has not been stated in AAS applications [27,28], but there has been some studies stating increases in AST and ALT activities [27,29-31].
In the study done, while hourly variation in any parameter has not been observed in male rats except GGT and BUN levels, this situation is also accompanied by ALP in females (Tables 4.1 and 4.2). However, when biochemical parameters are examined, there is numerical increase in especially AST and CK-MB levels in terms of h. Even if changes in biochemical parameters occurring in the first 24 h don't support increase in IL1-b and TNF-α levels completely, this has been thought as sign showing that ND applications will lead to various multiple organ failures particularly liver failure. As a matter of fact, although treatment was used in appropriate dose in a study in which ND was administered , with findings obtained at the end of 4 weeks, it was concluded that multiple organ failure might be encountered in rats.
As a result, it has been thought that these values are needed to be monitored longer than 24 h in order to understand changes in serum cytokine levels and biochemical parameters examined more clearly.
This study was supported by the scientific research coordinator SUBAPK (10401125). A part of the abstract was presented at the “13th International Congress of sport sciences” 07-9 November 2014, Konya,Turkey.
- Tayade MC, Bhamare SM, Kamble P, Jadhav K. Doping in sport: current review. Int J Cur Res Rev 2013; 7: 308-313.
- Beotra A. Drug abuse in sport: an on rewiev. J Postgrad Med Edu Res 2013; 47: 94-98.
- Bowers LD. Anti-dope testing in sport: the history and the science. FASEB J 2012; 26: 3933-3936.
- Thualagant N, Pfister G. The fihgt against fitness doping in sports clubs political discourses and strategies in Denmark. Performance Enhancement & Health 2012; 2:86-93.
- Bahrke MS, Yesalis CE. Abuse of anabolic androgenic steroids and related substances in sport and exercise. Curr Opin Pharmacol 2004; 4: 614-620.
- Thevis M, Schänzer W. Mass spectrometry in sports drug testing: Structure characterization and analytical assays. Mass Spectrom Rev 2007; 26: 79-107.
- Harmer PA. Anabolic-androgenic steroid use among young male and female athletes: is the game to blame? Br J Sports Med 2010; 44: 26-31.
- van Amsterdam J, Opperhuizen A, Hartgens F. Adverse health effects of anabolic-androgenic steroids. Regul Toxicol Pharmacol 2010; 57:117-123.
- Kerr JM, Congeni JA. Anabolic-androgenic steroids: use and abuse in pediatric patients. Pediatr Clin North Am 2007; 54: 771-785.
- Kuhn CM. Anabolic steroids. Recent Prog Horm Res 2002; 57: 411-434.
- Basualto-Alarcon C, Maass R, Jaimovich E, Estrada M. Anabolic/Androgenic steroid in skeletal muscle and cardiovasculer diseases. Itechopen 2013; 10:237-266.
- Donna LH, Jiaquan Xu. Deaths: Preliminary Data for 2011. National Vital Statistics Report 2012; 61: 1-52.
- Emtage LA, Trethowan C, Kelly K. A phase III open randomized study of Zoladex 3.6 mg depot versus DES 3 mg per day in untreated advanced prostate cancer: A West Midlands Urological Research Group Study. Prog Clin Biol Res 1989; 303: 47-52.
- D’Amico AV, Chen MH, Renshaw AA. Causes of death in men undergoing androgen suppression therapy for newly diagnosed localized or recurrent prostate cancer. Cancer 2008; 113: 3290-3297.
- Skogastierna C, Hotzen M1, Rane A1, Ekström L. A supraphysiological dose of testosterone induces nitric oxide production and oxidative stress. Eur J Prev Cardiol 2014; 21: 1049-1054.
- Evans NA. Current concepts in anabolic-androgenic steroids. Am J Sports Med 2004; 32: 534-542.
- Bruley-Rosset M, Dardenne M, Schuurs S. Functional and quantitave changes of immune vcells of ageing NZB mice treated with nandrolone decanoate. I. Effecct on survival and autoantibody development. Clin Exp Immunol 1985; 62: 630-638.
- Thompson RW, McClung JM, Baltgalvis KA, Davis JM, Carson JA. Modulation of overload-induced inflammation by aging and anabolic steroid administration. Exp Gerontol 2006; 41:1136-1148.
- Marshall-Gradisnik S, Green R, Brenu EW, Weatherby RP. Anabolic androgenic steroids effects on the immune system: a review. Cent Eur J Biol 2009; 4: 19-33.
- Hughes TK, Fulep E, Juelich T, Smith EM, Stanton GJ. Modulation of immune responses by anabolic androgenic steroids. Int J Immunopharmacol 1995; 17: 857-863.
- Saitoh T, Morimoto K, Kumagai T, Tsuboi I, Aikawa S, Horie T. Comparison of erythropoietic response to androgen in young and old senescense accelerated mice. Mech Ageing Dev 1999; 109: 125-139.
- Andreasen AS, Krabbe KS, Krogh-Madsen R, Taudorf S, Pedersen BK. Human endotoxemia as a model of systemic inflammation. Curr Med Chem 2008; 15: 1697-1705.
- Yamaguchi M, Weitzmann MN. The estrogen 17beta-estradiol and phytoestrogen genistein mediate differential effects on osteoblastic NF-KappaB activity. Int J Mol Med 2009; 23: 297-301.
- Corrales JJ, Almeida M, Burgo R. Androgen-replacement therapy depresses the ex vivo production of inflammatory cytokines by circulating antigen-presenting cells in aging type-2 diabetic men with partial androgen deficiency. J Endocrinol 2006; 189: 595-604.
- Parrillo JE, Parker MM, Natansnn C, Suffredini AF, Danner RL, Cunnion RE, Ognibene FP. Septic shock in humans. Advances in the understanding of pathogenesis, cardiovascular dysfunction, and therapy. Ann Intern Med 1990; 113: 227-242.
- Dinarello CA. Interleukin-1, interleukin-1 receptors and interleukin-1 receptor antagonist. Int Rev Immunol 1998; 16: 457-499.
- Alén M, Reinilä M, Vihko R. Response of serum hormones to androgen administration in power athletes. Med Sci Sports Exerc 1985; 17: 354-359.
- Kuipers H, Wijnen JA, Hartgens F, Willems SM. Influence of anabolic steroids on body composition, blood pressure, lipid profile and liver functions in body builders. Int J Sports Med 1991; 12: 413-418.
- Lenders JW, Demacker PN, Vos JA, Jansen PL, Hoitsma AJ, Van 't Laar A, Thien T. Deleterious effects of anabolic steroids on serum lipoproteins, blood pressure, and liver function in amateur body builders. Int J Sports Med 1988; 9: 19-23.
- Pertusi R, Dickerman RD, McConathy WJ. Evaluation of aminotranferase elevations in a bodybuilder using anabolic steroids: hepatitis or rhabdomyolysis? J Am Osteopath Assoc 2001; 101: 391-394.
- Urhausen A, Torsten A, Wilfrie K. Reversibility of the effects on blood cells, lipids, liver function and hormones in former anabolic–androgenic steroid abuser. J Steroid Biochem Mol Biol 2003; 84: 369-375.
- Tasgin E, Haliloglu S. The effect of nandrolone decanoate on the concentrations of IL1-ß, TNF-a, IL-4, IL-6 and biochemical parameters in male and female rats, PhD thesis, Selcuk Universty Health Sciences Institute 2013; 28-36.