Clinical Endocrinology (1 975) 4, 537-541.
THE EFFECTS O F L A R G E DOSES O F T H E A N A B O L I C STEROID, M E T H A N D R O S T E N O L O N E , O N A N ATHLETE B. H. K I L S H A W , R . A. H A R K N E S S , B. M. HOBSON A. W. M. S M I T H
AND
Departments of Paediatric Biochemistry, Clinical Chemistry, and Obstetrics and Gynaecology, University of Edinburgh, and Victoria Hospital, Kirkcaldy, Fqe (Accepted for publication I 6 December 1974)
SUMMARY
Doses of the anabolic steroid, methandrostenolone, nearly ten times greater than those used therapeutically, have resulted in a marked depression of levels of testosterone in urine and blood, and in some depression of gonadotrophin excretion by a male athlete. The administration of the drug has been checked and its metabolism studied from the pattern of urinary metabolites. The use of large doses of anabolic steroids by athletes to promote muscle development has been widely recognized but, understandably, little studied (Wade, 1972). The most popular drug is said to be methandrostenolone (Dianabol). In man, this steroid has been shown to increase weight (Wade, 1972) and induce nitrogen retention (Liddle & Burke, 1960). Endocrine studies have shown some alterations, in cortisol metabolism (James et al., 1962), in the production or release of ACTH (Wynn et al., 1962) and in the excretion of gonadotrophic activity (Almqvist et al., 1961). There is, however, little information on the effect of methandrostenolone or of other anabolic steroids on hypothalamic-pituitary-testicular function in man. This report shows an effect on an athlete of large doses of methandrostenolone on testosterone production and, to a lesser extent, gonadotrophin output. The metabolism of large doses of methandrostenolone appeared to differ from that of small doses. MATERIALS A N D METHODS The administration of methandrostenolone was checked and its urinary metabolites studied by the methods of Adhikary & Harkness (1969a, b). Testosterone in urine was measured by the method of Ismail & Harkness (1 966) modified by the inclusion of an internal radioactive standard for the determination of recovery. Plasma testosterone was measured by radioimmunoassay (Corker, 1974, personal communication). The gonadotrophins FSH and Correspondence: B. H. Kilshaw, Department of Paediatric Biochemistry, Royal Hospital for Sick Children, Edinburgh EH9 1LJ.
537
538
B. H. Kilshaw et al.
ICSH were measured in urine by the method of Schuurs & van Wyngaarden (1970). Using unconcentrated urine, this method can measure levels of FSH from 0.5 iu/l and of ICSH from 12 iu/l. Urinary pregnanetriol and 17-hydroxycorticosteroids (17-OHCS) were measured by the methods of Harkness & Love (1966) and Few (1960) respectively. RESULTS
Case report: clinical A 26-year-old man started taking methandrostenolone in 1971 to help him gain financial rewards in weight-lifting and body-building competitions. He combined a dose of up to 100 mg/24 h of methandrostenolone with physical training. His weight subsequently increased by 12.7 kg to 95 kg over a period of about 2 years, while his muscular development was spectacular. Initially his married life was normal; there was no family despite the use of the unreliable ‘safe period’. In 1973 some problems in this area prompted him to seek medical advice and in the course of the original and subsequent investigation the results presented here were obtained. When this man reappeared 28 weeks after the initial consultation, no obvious abnormality was present on definitive physical examination which included testicular size and consistency. His sperm counts were somewhat low (McLeod, 1951) and at this time were: Total count Motility Abnormal Date Vol. (ml) x 106/ml (%I forms (%) 9.4.74 1.1 45 35 20 50 16.4.74 2.0 40.5 40 18.4.74 2.0 15.7 40 30 Endocrine studies Studies of his excretion of testosterone and gonadotrophins are shown in Fig. 1. There was a marked depression of urinary testosterone to almost castrate levels (Ismail & Harkness, 1966) and there was a similar depression of plasma testosterone to a level of 67.8 ng/dl in week 28 (n = 5). ICSH and FSH levels in urine were in the low normal range for males. FSH levels appeared relatively lower than those of ICSH. The mean urinary outputs ( 5 S E M ) of normal men were 35.55 1.98 (n = 49) and 4.7A 1.16 (n = 22) for ICSH and FSH respectively. Both gonadotrophin and testosterone excretion rose markedly when administration of methandrostenolone ceased. A more marked rise was noted in ICSH than in FSH levels. Testosterone output was, however, still subnormal. After restarting selfmedication with methandrostenolone and gradually increasing the dosage, the subject’s testosterone excretion had fallen in week 28 to its previous low level, while his FSH output was also lower. Pregnanetriol excretion during this week was subnormal at 0.1 1 and 0.14 mg/24 h ; this is consistent with the low testosterone levels observed (Harkness & Love, 1966). Urinary 17-OHCS were in the lower part of the normal range and possibly showed some increase on stopping self-medication (Wynn et al., 1962). The administration of methandrostenolone was checked and its metabolism studied from the output of metabolites in urine. The ‘total’ recoveries of the stated dose ranged from 1.5 to 6%, with the exception of weeks 3 and 4 when he was reducing his intake. At this time the excretion was about 14% of the dose, presumably due to a delay in the excretion.
Efect of an anabolic steroid on an athlete
539
:h,
5r
0.2
0
r LL v)
' M D I
0
6o
rI-
NLd I
;h ik-lj-l, 0
g
20
0
I
0
1
2
3
4
5
+ 28
Weeks
28
Weeks
FIG.1. The urinary excretion of the gonadotrophins, FSH and ICSH, testosterone, 17-hydroxycorticosteroids and creatinine by a male athlete taking the indicated large doses of methandrostenolone. The urinary output of the two major metabolites of this anabolic steriod, the 68-hydroxy and 17-epimer compounds are also shown.
540
B. H. Kilshaw et al.
These ‘total’ recoveries are similar to those obtained from the normal therapeutic dose (Harkness et al., 1974). However, the amount of the 17-epimer is greatly increased relative to the amount of 6P-hydroxy metabolite. The effects of these large doses of the anabolic steroid were shown by a raised creatinine excretion being 3.25, 3.19, 2-65, 2.69 and 3.75 g/24 h in weeks 1-5 respectively. DISCUSSION The dose of methandrostenolone for full androgen substitution is about 15 mgj24 h for adult males (Liddle & Burke, 1960). It is not surprising, therefore, that testosterone output in this case has been depressed by the large doses of this anabolic steroid to virtually castrate levels. This is consistent with the observed depression of the weight of the testes in rats after methandrostenolone administration (Boris et al., 1969). However, the gonadotrophins are not suppressed to the extent which would be expected from the testosterone results. It is therefore possible that these very large doses may act partly by direct inhibition of testicular steroid biosynthesis as well as by central effects. There is clear evidence of androgenic and anabolic effects in this man. However, his creatinine excretion did not appear to be clearly correlated with dose. The observations of Liddle & Burke (1960) that weight increase plateaued at a dose of 2.5 mg/24 h, and those of Kruskemper (1968) that nitrogen retention in healthy men did not continue for more than a few weeks despite an increase in dosage, also suggest that large doses are not proportionately effective in building muscle in man despite the evidence from rats (Desaulles et al., 1959). The effects of methandrostenolone on liver function are varied. BSP retention has been shown to increase linearly with dose (Liddle & Burke, 1960). The present results are consistent with an alteration in the metabolism of this drug with increasing dosage, and suggest that the capacity of the liver for 6P-hydroxylation of steroids is limited. The increase in the plasma half-life of cortisol after methandrostenolone (James et al., 1962) is also consistent with this conclusion since both methandrostenolone and cortisol undergo 6P-hydroxylation (Harkness et al., 1974; Southren et al., 1969). The ability for 17-epimerization of the drug is high in this man. In conclusion, the use of an anabolic steroid in amounts nearly ten times greater than the normal therapeutic dose has resulted in marked depression of testosterone with some depression of gonadotrophin excretion. There is also some evidence of dose-related alterations in hepatic function. These studies would appear to provide evidence against the use of large doses of at least one anabolic steroid by normal men. ACKNOWLEDGMENTS
We are grateful to Dr D. D. Kennedy, Dr C. S. Corker and Dr D. M. Burley for their great help with this study. REFERENCES ADHIKARY, P.M. & HARKNESS, R.A. (1969a) Determination of the carbon skeletons of microgram amounts of steroids and sterols by gas chromatography after their high temperature catalytic reduction. Analytical Chemistry, 41, 410. ADHIKARY, P.M. & HARKNESS, R.A. (1969b) Production of the parent hydrocarbons from steroid drugs and their separation by gas chromatography. Journal of Chromatography, 42,29.
Eflect of an anabolic steroid on an athlete
541
ALMQVIST, S., IKKOS, D. & LUFT, R. (1961) Metabolic studies with methandrostenolone (17&hydroxy-l7amethyl-androsta-l,4-dien-3-one) in human subjects. Acta Endocrinologica, 38, 413. T. (1969) Comparative androgenic, myotrophic and antigonadotroBORIS,A., STEVENSON, R.H. & TRMAL, phic properties of some anabolic steroids. Steroids, 15, 61. DESAULLES, P.A., KRAKENBUHL, CH., SCHULER, W. & BEIN,H.J. (1959) Experimental study on Dianabol, a new anabolic agent. Schweizerische Medizinische Wochenschrijt, 89, 1313. FEW,J.D. (1960) A method for the analysis of urinary 17-hydroxycorticosteroids.Journal of Endocrinology, 22, 31. HARKNESS, R.A. & LOVE,D.N. (1966) Studies on the estimation of urinary pregnanetriol during pregnancy and childhood. Acta Endocrinologica, 51, 526. J.A. (1974) Physiological and pharmacological factors affecting HARKNESS, R.A., SCOTT,R.D.M. & STRONG, the 68-hydroxylation and 17-epimerization of methandrostenolone. Biochemical Society Transactions, 2, 119. R.A. (1966) A method for the estimation of urinary testosterone. Biochemical ISMAIL,A.A.A. & HARKNESS, Journal, 99, 717. JAMES, V.H.T., LANDON, J. & WYNN,V. (1962) Effect of an anabolic steroid (methandienone) on the metabolism of cortisol in the human. Journal of Endocrinology, 25,211. KRUSKEMPER, H.L. (1968) Anabolic Steroids. Academic Press, New York. LIDDLE,G.W. & BURKE,H.A. (1960) Anabolic steroids in clinical medicine. Helvetica Medica Acta, 27, 504. MCLEOD,J. (1951) Semen quality in one thousand men of known fertility and in eight hundred cases of infertile marriages. Fertility and Sterility, 2, 115. SCHUURS, A.H.W.M. & VAN WYNGAARDEN, C.J. (1970) Modified haemagglutination test and its application for the estimation of LH in unconcentrated urine. Acta Endocrinologica, Suppl. 141, 13. SOUTHREN, A.L., GORDON,G.G., TOCHIMOTO, S., KRIKUN,E., KRIEGER, D., JACOBSON, M. & KUNTZMAN, R. (1969) Effect of N-phenylbarbital (phetharbital) on the metabolism of testosterone and cortisol in man. Journal of Clinical Endocrinology, 29,251. WADE,N. (1972) Anabolic steroids: doctors denounce them, but athletes aren’t listening. Science, 176,1399. J. & JAMES, V.H.T. (1962) Effect of an anabolic steroid (methandienone) on pituitaryWYNN,V., LANDON, adrenal function in the human. Journal of Endocrinology, 25, 199.
THE EFFECTS O F L A R G E DOSES O F T H E A N A B O L I C STEROID, M E T H A N D R O S T E N O L O N E , O N A N ATHLETE B. H. K I L S H A W , R . A. H A R K N E S S , B. M. HOBSON A. W. M. S M I T H
AND
Departments of Paediatric Biochemistry, Clinical Chemistry, and Obstetrics and Gynaecology, University of Edinburgh, and Victoria Hospital, Kirkcaldy, Fqe (Accepted for publication I 6 December 1974)
SUMMARY
Doses of the anabolic steroid, methandrostenolone, nearly ten times greater than those used therapeutically, have resulted in a marked depression of levels of testosterone in urine and blood, and in some depression of gonadotrophin excretion by a male athlete. The administration of the drug has been checked and its metabolism studied from the pattern of urinary metabolites. The use of large doses of anabolic steroids by athletes to promote muscle development has been widely recognized but, understandably, little studied (Wade, 1972). The most popular drug is said to be methandrostenolone (Dianabol). In man, this steroid has been shown to increase weight (Wade, 1972) and induce nitrogen retention (Liddle & Burke, 1960). Endocrine studies have shown some alterations, in cortisol metabolism (James et al., 1962), in the production or release of ACTH (Wynn et al., 1962) and in the excretion of gonadotrophic activity (Almqvist et al., 1961). There is, however, little information on the effect of methandrostenolone or of other anabolic steroids on hypothalamic-pituitary-testicular function in man. This report shows an effect on an athlete of large doses of methandrostenolone on testosterone production and, to a lesser extent, gonadotrophin output. The metabolism of large doses of methandrostenolone appeared to differ from that of small doses. MATERIALS A N D METHODS The administration of methandrostenolone was checked and its urinary metabolites studied by the methods of Adhikary & Harkness (1969a, b). Testosterone in urine was measured by the method of Ismail & Harkness (1 966) modified by the inclusion of an internal radioactive standard for the determination of recovery. Plasma testosterone was measured by radioimmunoassay (Corker, 1974, personal communication). The gonadotrophins FSH and Correspondence: B. H. Kilshaw, Department of Paediatric Biochemistry, Royal Hospital for Sick Children, Edinburgh EH9 1LJ.
537
538
B. H. Kilshaw et al.
ICSH were measured in urine by the method of Schuurs & van Wyngaarden (1970). Using unconcentrated urine, this method can measure levels of FSH from 0.5 iu/l and of ICSH from 12 iu/l. Urinary pregnanetriol and 17-hydroxycorticosteroids (17-OHCS) were measured by the methods of Harkness & Love (1966) and Few (1960) respectively. RESULTS
Case report: clinical A 26-year-old man started taking methandrostenolone in 1971 to help him gain financial rewards in weight-lifting and body-building competitions. He combined a dose of up to 100 mg/24 h of methandrostenolone with physical training. His weight subsequently increased by 12.7 kg to 95 kg over a period of about 2 years, while his muscular development was spectacular. Initially his married life was normal; there was no family despite the use of the unreliable ‘safe period’. In 1973 some problems in this area prompted him to seek medical advice and in the course of the original and subsequent investigation the results presented here were obtained. When this man reappeared 28 weeks after the initial consultation, no obvious abnormality was present on definitive physical examination which included testicular size and consistency. His sperm counts were somewhat low (McLeod, 1951) and at this time were: Total count Motility Abnormal Date Vol. (ml) x 106/ml (%I forms (%) 9.4.74 1.1 45 35 20 50 16.4.74 2.0 40.5 40 18.4.74 2.0 15.7 40 30 Endocrine studies Studies of his excretion of testosterone and gonadotrophins are shown in Fig. 1. There was a marked depression of urinary testosterone to almost castrate levels (Ismail & Harkness, 1966) and there was a similar depression of plasma testosterone to a level of 67.8 ng/dl in week 28 (n = 5). ICSH and FSH levels in urine were in the low normal range for males. FSH levels appeared relatively lower than those of ICSH. The mean urinary outputs ( 5 S E M ) of normal men were 35.55 1.98 (n = 49) and 4.7A 1.16 (n = 22) for ICSH and FSH respectively. Both gonadotrophin and testosterone excretion rose markedly when administration of methandrostenolone ceased. A more marked rise was noted in ICSH than in FSH levels. Testosterone output was, however, still subnormal. After restarting selfmedication with methandrostenolone and gradually increasing the dosage, the subject’s testosterone excretion had fallen in week 28 to its previous low level, while his FSH output was also lower. Pregnanetriol excretion during this week was subnormal at 0.1 1 and 0.14 mg/24 h ; this is consistent with the low testosterone levels observed (Harkness & Love, 1966). Urinary 17-OHCS were in the lower part of the normal range and possibly showed some increase on stopping self-medication (Wynn et al., 1962). The administration of methandrostenolone was checked and its metabolism studied from the output of metabolites in urine. The ‘total’ recoveries of the stated dose ranged from 1.5 to 6%, with the exception of weeks 3 and 4 when he was reducing his intake. At this time the excretion was about 14% of the dose, presumably due to a delay in the excretion.
Efect of an anabolic steroid on an athlete
539
:h,
5r
0.2
0
r LL v)
' M D I
0
6o
rI-
NLd I
;h ik-lj-l, 0
g
20
0
I
0
1
2
3
4
5
+ 28
Weeks
28
Weeks
FIG.1. The urinary excretion of the gonadotrophins, FSH and ICSH, testosterone, 17-hydroxycorticosteroids and creatinine by a male athlete taking the indicated large doses of methandrostenolone. The urinary output of the two major metabolites of this anabolic steriod, the 68-hydroxy and 17-epimer compounds are also shown.
540
B. H. Kilshaw et al.
These ‘total’ recoveries are similar to those obtained from the normal therapeutic dose (Harkness et al., 1974). However, the amount of the 17-epimer is greatly increased relative to the amount of 6P-hydroxy metabolite. The effects of these large doses of the anabolic steroid were shown by a raised creatinine excretion being 3.25, 3.19, 2-65, 2.69 and 3.75 g/24 h in weeks 1-5 respectively. DISCUSSION The dose of methandrostenolone for full androgen substitution is about 15 mgj24 h for adult males (Liddle & Burke, 1960). It is not surprising, therefore, that testosterone output in this case has been depressed by the large doses of this anabolic steroid to virtually castrate levels. This is consistent with the observed depression of the weight of the testes in rats after methandrostenolone administration (Boris et al., 1969). However, the gonadotrophins are not suppressed to the extent which would be expected from the testosterone results. It is therefore possible that these very large doses may act partly by direct inhibition of testicular steroid biosynthesis as well as by central effects. There is clear evidence of androgenic and anabolic effects in this man. However, his creatinine excretion did not appear to be clearly correlated with dose. The observations of Liddle & Burke (1960) that weight increase plateaued at a dose of 2.5 mg/24 h, and those of Kruskemper (1968) that nitrogen retention in healthy men did not continue for more than a few weeks despite an increase in dosage, also suggest that large doses are not proportionately effective in building muscle in man despite the evidence from rats (Desaulles et al., 1959). The effects of methandrostenolone on liver function are varied. BSP retention has been shown to increase linearly with dose (Liddle & Burke, 1960). The present results are consistent with an alteration in the metabolism of this drug with increasing dosage, and suggest that the capacity of the liver for 6P-hydroxylation of steroids is limited. The increase in the plasma half-life of cortisol after methandrostenolone (James et al., 1962) is also consistent with this conclusion since both methandrostenolone and cortisol undergo 6P-hydroxylation (Harkness et al., 1974; Southren et al., 1969). The ability for 17-epimerization of the drug is high in this man. In conclusion, the use of an anabolic steroid in amounts nearly ten times greater than the normal therapeutic dose has resulted in marked depression of testosterone with some depression of gonadotrophin excretion. There is also some evidence of dose-related alterations in hepatic function. These studies would appear to provide evidence against the use of large doses of at least one anabolic steroid by normal men. ACKNOWLEDGMENTS
We are grateful to Dr D. D. Kennedy, Dr C. S. Corker and Dr D. M. Burley for their great help with this study. REFERENCES ADHIKARY, P.M. & HARKNESS, R.A. (1969a) Determination of the carbon skeletons of microgram amounts of steroids and sterols by gas chromatography after their high temperature catalytic reduction. Analytical Chemistry, 41, 410. ADHIKARY, P.M. & HARKNESS, R.A. (1969b) Production of the parent hydrocarbons from steroid drugs and their separation by gas chromatography. Journal of Chromatography, 42,29.
Eflect of an anabolic steroid on an athlete
541
ALMQVIST, S., IKKOS, D. & LUFT, R. (1961) Metabolic studies with methandrostenolone (17&hydroxy-l7amethyl-androsta-l,4-dien-3-one) in human subjects. Acta Endocrinologica, 38, 413. T. (1969) Comparative androgenic, myotrophic and antigonadotroBORIS,A., STEVENSON, R.H. & TRMAL, phic properties of some anabolic steroids. Steroids, 15, 61. DESAULLES, P.A., KRAKENBUHL, CH., SCHULER, W. & BEIN,H.J. (1959) Experimental study on Dianabol, a new anabolic agent. Schweizerische Medizinische Wochenschrijt, 89, 1313. FEW,J.D. (1960) A method for the analysis of urinary 17-hydroxycorticosteroids.Journal of Endocrinology, 22, 31. HARKNESS, R.A. & LOVE,D.N. (1966) Studies on the estimation of urinary pregnanetriol during pregnancy and childhood. Acta Endocrinologica, 51, 526. J.A. (1974) Physiological and pharmacological factors affecting HARKNESS, R.A., SCOTT,R.D.M. & STRONG, the 68-hydroxylation and 17-epimerization of methandrostenolone. Biochemical Society Transactions, 2, 119. R.A. (1966) A method for the estimation of urinary testosterone. Biochemical ISMAIL,A.A.A. & HARKNESS, Journal, 99, 717. JAMES, V.H.T., LANDON, J. & WYNN,V. (1962) Effect of an anabolic steroid (methandienone) on the metabolism of cortisol in the human. Journal of Endocrinology, 25,211. KRUSKEMPER, H.L. (1968) Anabolic Steroids. Academic Press, New York. LIDDLE,G.W. & BURKE,H.A. (1960) Anabolic steroids in clinical medicine. Helvetica Medica Acta, 27, 504. MCLEOD,J. (1951) Semen quality in one thousand men of known fertility and in eight hundred cases of infertile marriages. Fertility and Sterility, 2, 115. SCHUURS, A.H.W.M. & VAN WYNGAARDEN, C.J. (1970) Modified haemagglutination test and its application for the estimation of LH in unconcentrated urine. Acta Endocrinologica, Suppl. 141, 13. SOUTHREN, A.L., GORDON,G.G., TOCHIMOTO, S., KRIKUN,E., KRIEGER, D., JACOBSON, M. & KUNTZMAN, R. (1969) Effect of N-phenylbarbital (phetharbital) on the metabolism of testosterone and cortisol in man. Journal of Clinical Endocrinology, 29,251. WADE,N. (1972) Anabolic steroids: doctors denounce them, but athletes aren’t listening. Science, 176,1399. J. & JAMES, V.H.T. (1962) Effect of an anabolic steroid (methandienone) on pituitaryWYNN,V., LANDON, adrenal function in the human. Journal of Endocrinology, 25, 199.
