andrologia 7 ( 3 ) : 195-198
(1975)
Received October 7, 1974
Biomedical Division, National Institute of Family Planning, New DeIhi/India
Effect of androgen on different biochemical constituents of human semen S. ROY, R.P. DAS and S.L. TANEJA
Various biochemical constituents of the semen are contributed by different accessory sex organs, which are androgen dependent (Dorfman and Shipley - 1956; Price and Williams-Ashman- 1961 ;Mann - 1964). Although some investigators have reported on the effect of androgen on the level of fructose (Tyler et al. - 1955) and the acid phosphatase activity (El-Ridi et al. - 1962) in human semen, no information is available on the relative sensitivty of different biochemical parameters to androgen administration. The present report embodies the results of a study on the effect of androgen admini stration on the levels of fructose, cholesterol, ascorbic acid and sialic acid and the activities of acid and alkaline phosphatases in eight healthy volunteers with non-obstructive azoospermia. Materials and Methods Eight healthy volunteers who attended the Infertility Clinic of the Institute with azoospermia resulting from spermatogenic defect, as revealed by testicular biopsy, were enrolled for this study. After obtaining two control semen samples, each individual was administered intramuscularly 50 mg. of testosterone propionate thrice weekly for 4 weeks. Semen samples were collected in dry and clean glass containers by masturbation after each three injections. Three days abstinence was observed before collection of each sample, and all samples were processed for biochemical estimations. The levels of fructose, cholesterol, ascorbic acid and sialic acid, and the activities of the acid and alkaline phosphatases were determined according to the methods of Mann (1964), van Botzelaer and Zondag (1960), Roe and Kuether (1943), Warren (19591, Gutman and Gutman (1940), and King and Armstrong (1934) respectively. Results The results are presented in fig. 1. The activity of acid phosphatase increased progressively and was significantly elevated after the sixth injection. The concentration of fructose also increased progressively with androgen administration, although the magnitude of rise was relatively less and the difference from the control was statistically significant only after the ninth injection. The levels of sialic acid as well as the alkaline phosphatase activity showed elevations which became significant only after twelve injections of androgen. The cholesterol as well as ascorbic acid contents of the semen did not show any significant change even at the end of twelve injections. Key words: Androgen - biochemistry - human semen phosphatase sialic acid - cholesterol - ascorbic acid ~
-
fructose - alk. phosphatase
-
acid
S. ROY,R. P. DASand S. L. TANEJA
196
d 1400
9
9
1200
A
x
300
,1 200
Lr
100
C T Ta TI T.
I2Ol
0
T
E
’3
r
6 16
‘4
14 12
0
10
C, b Ca. TWO CONTROL VALUES
T, Ta T,IT4mVALUES AFTER 1,2,364
WEEKd TRLATMENT
Fig.: Levels (mean f S.E.) of acid and alkaline phosphatases, fructose, sialic acid, cholesterol and ascorbic acid in human semen before and during administration of testosterone propionate over a four weeks period.
Discussion
Of the different seminal constituents studied acid phosphatase appeared to be the most sensitive, followed in order of responsiveness by fructose, sialic acid and alkaline phosphatase. The levels of cholesterol and ascorbic acid did not change even after 12 Iniections of 50 mg. of testosterone propionate given over a period of 4 weeks. The dose of testosterone used was high enough to suppress spermatogenesis (Reddy and Rao - 1972). El-Ridi et al. (1 962) reported that seminal acid phosphatase activity was low in azoospermic individuals and there was almost a 10 fold increase in its activity following testosterone administration. Mann and Parson (1947) showed in rabbit that within 2 weeks after castration seminal fructose disappeared almost completely; implantation of testosterone pellet could prevent as well as restore this. The possibility of assessing the androgenic secretion of male gonads on the basis of fructose levels in seminal fluid has been suggested (Davies and Mann - 1947; Landau and Loughead - 1951). Tyler (19550, however, detected only moderate increase in seminal fructose level in 2 out andrologia 7, Heft 3 (1975)
Effect of androgen on different biochemical constituents of human semen
197
of 6 subjects who received daily 50 mg. of testosterone propionate parenterally. He felt that small amount of endogenous testosterone might be sufficient to maintain normal range of fructose in the semen and additional amounts did not alter this function much. In a more recent study on 47 normal and pathological subjects, Dondero et al. (1970) did not find any significant correlation between the levels of plasma testosterone and seminal fructose. They concluded that fructose assay in seminal plasma could not afford any index of androgen secretion from the male gonad. Similar information in regard to the other biochemical constituents of the human semen is not available. Nevertheless, from the results of the present study in azoospermic individuals it appears that pharmacological doses of testosterone are needed to elevate the levels of some of these constituents, and seminal cholesterol and ascorbic acid are refractory even to such high dose of androgen. Summary Changes in the levels of fructose, sialic acid, ascorbic acid, cholesterol and acid and alkaline phosphatases in the semen were studied in eight azoospermic volunteers during administration of testosterone propionate over a four week period. In order of responsiveness, elevations were observed in the levels of acid phosphatase, fructose, sialic acid and alkaline phosphatase during androgen therapy. No changes were seen in the levels of cholesterol and ascorbic acid.
Der EinfluR von Androgenen auf verschiedene biochemische Bestandteile des menschlichen Spermas
Zusammenfassung In einer experimentellen Arbeit wird zur Beeinflussung von Fruktose, Sialinsaure, Ascorbinsaure, Cholesterin und der alk. Phosphatase bei Azoospermie-Patienten unter einer Behandlung mit Testosteronpropionat (50 mg dreimal wochentlich iiber 4 Wochen bei 8 Mannern) untersucht. Es kann gezeigt werden, dai3 bei den Werten fur alk. Phosphatase, Fruktose, Sialinsare und saure Phosphatase ein Anstieg zu beobachten ist, wahrend fur Cholesterin und Ascorbinsaure keinerlei Veranderungen festzustellen waren.
Efectos de androgenos sobre componentes bioquimicos del semen humano
Resumen En ocho voluntarios azoospkrmicos, se estudiaron 10s cambios en semen de 10s niveles de fructosa, Bcido siilico, Bcido ascbrbico, colesterol y fosfatasas Bcidas y alcah a s , durante la administracibn de propionato de testosterona, durante un periode de 4 semanas. Se observaron elevaciones de 10s niveles de fosfatasas Bcidas y alcalinas, fructosa y k i d 0 siilico durante el tratamiento androghico. No se observaron cambios en 10s niveles de colesterol y Bcido ascbrbico. andrologia 7,Heft 3 (1975)
198
S. Roy, R. P. DASand S. L. TANEJA
References Boetzelaer, G.L. van and H.A. Zondag: A rapid modification of the Pearson reaction for total serum cholesterol. Clin. Chim. Acta 5, 943 (1960). Conor, W.E.: The transfer of (4-14C) cholesterol from blood to semen in man. J. Reprod. Fertil. 17, 211-13 (1968). Davies, D.V. and T. Mann: Functional development of accessory glands and spermatogenesis. Nature 160, 205-206 (1947). Dondero, F., F. Sciarra and A. Isidori: Statistical evaluation of relationships between plasma testosterone and seminal fructose. Folia Endocrinol. 23, 559-565 (1970). Dorfman, R. and R.A. Shipley: Androgens: Biochemistry, Physiology and Clinical Significance. J. Wiley and Sons, New York (19561, p. 152-186. Eliasson, R.: Effect of frequent ejaculations on the composition of human seminal plasma. J . Reprod. Fertil. 9, 331-336 (1965). El-Ridi, M.S., L. Soliman, A. El-Etreby and A. Ibrahim: Egypt Pharm. Bull. 44,355-361 (1962). Gutman, E.B. and A.B. Gutman: Estimation of acid phosphatase activity of blood serum. J. Biol. Chem. 136, 201-209 (1940). King, E.J. and A.R. Armstrong: Convenient method for determining serum and bile phosphatase activity. Canad. Med. Assoc. J. 31, 376 (1934). Landau, R.L. and R. Loughead: Seminal fructose as index of androgenic activity in man. J . Clin. Endocr. 11,1411-1424 (1951). Mann, T.: Biochemistry of semen and of male reproductive tract. John Wiley, New York (1964). Mann, T. and U. Parsons: Effect of testicular hormone on the formation of seminal fructose. Nature, 160, 294 (1947). Price, D. and H.G. Williams-Ashman: The accessory reproductive glands of mammals. In Young, W. C. and Corner, G.W. (Eds.). Sex and Internal secretions. The Williams and Wilkins Co., Baltimore (1961), p. 366-448. Reddy, P.R.K. and J.M. Rao: Reversible antifertility action of testosterone propionate in human males. Contraception 5, 295-301 (1972). Roe, J.H. and C.A. Kuether: Determination of ascorbic acid in whole blood and urine. T.. Biol. Chem. 147, 399-407 (1943). Tyler, E.T.: Seminal fructose studies in infertility. Fertil. Steril. 6, 247-258 (1955). Warren, L., Thiobarbituric acid assay of sialic acids. J. Biol. Chem. 234, 1971-1975 (1959).
Acknowledgement: The authors wish t o thank Dr. R.K. Sanyal for his interest in this study. Skillful laboratory assistance of Shri H.C. Pandey is acknowledged.
Address: Dr. Somnath ROY, Biomedical Division, National Institute of Family Planning, D-5, Green Park, New Delhi-110016, India.
andrologia 7, Heft 3 (1975)
(1975)
Received October 7, 1974
Biomedical Division, National Institute of Family Planning, New DeIhi/India
Effect of androgen on different biochemical constituents of human semen S. ROY, R.P. DAS and S.L. TANEJA
Various biochemical constituents of the semen are contributed by different accessory sex organs, which are androgen dependent (Dorfman and Shipley - 1956; Price and Williams-Ashman- 1961 ;Mann - 1964). Although some investigators have reported on the effect of androgen on the level of fructose (Tyler et al. - 1955) and the acid phosphatase activity (El-Ridi et al. - 1962) in human semen, no information is available on the relative sensitivty of different biochemical parameters to androgen administration. The present report embodies the results of a study on the effect of androgen admini stration on the levels of fructose, cholesterol, ascorbic acid and sialic acid and the activities of acid and alkaline phosphatases in eight healthy volunteers with non-obstructive azoospermia. Materials and Methods Eight healthy volunteers who attended the Infertility Clinic of the Institute with azoospermia resulting from spermatogenic defect, as revealed by testicular biopsy, were enrolled for this study. After obtaining two control semen samples, each individual was administered intramuscularly 50 mg. of testosterone propionate thrice weekly for 4 weeks. Semen samples were collected in dry and clean glass containers by masturbation after each three injections. Three days abstinence was observed before collection of each sample, and all samples were processed for biochemical estimations. The levels of fructose, cholesterol, ascorbic acid and sialic acid, and the activities of the acid and alkaline phosphatases were determined according to the methods of Mann (1964), van Botzelaer and Zondag (1960), Roe and Kuether (1943), Warren (19591, Gutman and Gutman (1940), and King and Armstrong (1934) respectively. Results The results are presented in fig. 1. The activity of acid phosphatase increased progressively and was significantly elevated after the sixth injection. The concentration of fructose also increased progressively with androgen administration, although the magnitude of rise was relatively less and the difference from the control was statistically significant only after the ninth injection. The levels of sialic acid as well as the alkaline phosphatase activity showed elevations which became significant only after twelve injections of androgen. The cholesterol as well as ascorbic acid contents of the semen did not show any significant change even at the end of twelve injections. Key words: Androgen - biochemistry - human semen phosphatase sialic acid - cholesterol - ascorbic acid ~
-
fructose - alk. phosphatase
-
acid
S. ROY,R. P. DASand S. L. TANEJA
196
d 1400
9
9
1200
A
x
300
,1 200
Lr
100
C T Ta TI T.
I2Ol
0
T
E
’3
r
6 16
‘4
14 12
0
10
C, b Ca. TWO CONTROL VALUES
T, Ta T,IT4mVALUES AFTER 1,2,364
WEEKd TRLATMENT
Fig.: Levels (mean f S.E.) of acid and alkaline phosphatases, fructose, sialic acid, cholesterol and ascorbic acid in human semen before and during administration of testosterone propionate over a four weeks period.
Discussion
Of the different seminal constituents studied acid phosphatase appeared to be the most sensitive, followed in order of responsiveness by fructose, sialic acid and alkaline phosphatase. The levels of cholesterol and ascorbic acid did not change even after 12 Iniections of 50 mg. of testosterone propionate given over a period of 4 weeks. The dose of testosterone used was high enough to suppress spermatogenesis (Reddy and Rao - 1972). El-Ridi et al. (1 962) reported that seminal acid phosphatase activity was low in azoospermic individuals and there was almost a 10 fold increase in its activity following testosterone administration. Mann and Parson (1947) showed in rabbit that within 2 weeks after castration seminal fructose disappeared almost completely; implantation of testosterone pellet could prevent as well as restore this. The possibility of assessing the androgenic secretion of male gonads on the basis of fructose levels in seminal fluid has been suggested (Davies and Mann - 1947; Landau and Loughead - 1951). Tyler (19550, however, detected only moderate increase in seminal fructose level in 2 out andrologia 7, Heft 3 (1975)
Effect of androgen on different biochemical constituents of human semen
197
of 6 subjects who received daily 50 mg. of testosterone propionate parenterally. He felt that small amount of endogenous testosterone might be sufficient to maintain normal range of fructose in the semen and additional amounts did not alter this function much. In a more recent study on 47 normal and pathological subjects, Dondero et al. (1970) did not find any significant correlation between the levels of plasma testosterone and seminal fructose. They concluded that fructose assay in seminal plasma could not afford any index of androgen secretion from the male gonad. Similar information in regard to the other biochemical constituents of the human semen is not available. Nevertheless, from the results of the present study in azoospermic individuals it appears that pharmacological doses of testosterone are needed to elevate the levels of some of these constituents, and seminal cholesterol and ascorbic acid are refractory even to such high dose of androgen. Summary Changes in the levels of fructose, sialic acid, ascorbic acid, cholesterol and acid and alkaline phosphatases in the semen were studied in eight azoospermic volunteers during administration of testosterone propionate over a four week period. In order of responsiveness, elevations were observed in the levels of acid phosphatase, fructose, sialic acid and alkaline phosphatase during androgen therapy. No changes were seen in the levels of cholesterol and ascorbic acid.
Der EinfluR von Androgenen auf verschiedene biochemische Bestandteile des menschlichen Spermas
Zusammenfassung In einer experimentellen Arbeit wird zur Beeinflussung von Fruktose, Sialinsaure, Ascorbinsaure, Cholesterin und der alk. Phosphatase bei Azoospermie-Patienten unter einer Behandlung mit Testosteronpropionat (50 mg dreimal wochentlich iiber 4 Wochen bei 8 Mannern) untersucht. Es kann gezeigt werden, dai3 bei den Werten fur alk. Phosphatase, Fruktose, Sialinsare und saure Phosphatase ein Anstieg zu beobachten ist, wahrend fur Cholesterin und Ascorbinsaure keinerlei Veranderungen festzustellen waren.
Efectos de androgenos sobre componentes bioquimicos del semen humano
Resumen En ocho voluntarios azoospkrmicos, se estudiaron 10s cambios en semen de 10s niveles de fructosa, Bcido siilico, Bcido ascbrbico, colesterol y fosfatasas Bcidas y alcah a s , durante la administracibn de propionato de testosterona, durante un periode de 4 semanas. Se observaron elevaciones de 10s niveles de fosfatasas Bcidas y alcalinas, fructosa y k i d 0 siilico durante el tratamiento androghico. No se observaron cambios en 10s niveles de colesterol y Bcido ascbrbico. andrologia 7,Heft 3 (1975)
198
S. Roy, R. P. DASand S. L. TANEJA
References Boetzelaer, G.L. van and H.A. Zondag: A rapid modification of the Pearson reaction for total serum cholesterol. Clin. Chim. Acta 5, 943 (1960). Conor, W.E.: The transfer of (4-14C) cholesterol from blood to semen in man. J. Reprod. Fertil. 17, 211-13 (1968). Davies, D.V. and T. Mann: Functional development of accessory glands and spermatogenesis. Nature 160, 205-206 (1947). Dondero, F., F. Sciarra and A. Isidori: Statistical evaluation of relationships between plasma testosterone and seminal fructose. Folia Endocrinol. 23, 559-565 (1970). Dorfman, R. and R.A. Shipley: Androgens: Biochemistry, Physiology and Clinical Significance. J. Wiley and Sons, New York (19561, p. 152-186. Eliasson, R.: Effect of frequent ejaculations on the composition of human seminal plasma. J . Reprod. Fertil. 9, 331-336 (1965). El-Ridi, M.S., L. Soliman, A. El-Etreby and A. Ibrahim: Egypt Pharm. Bull. 44,355-361 (1962). Gutman, E.B. and A.B. Gutman: Estimation of acid phosphatase activity of blood serum. J. Biol. Chem. 136, 201-209 (1940). King, E.J. and A.R. Armstrong: Convenient method for determining serum and bile phosphatase activity. Canad. Med. Assoc. J. 31, 376 (1934). Landau, R.L. and R. Loughead: Seminal fructose as index of androgenic activity in man. J . Clin. Endocr. 11,1411-1424 (1951). Mann, T.: Biochemistry of semen and of male reproductive tract. John Wiley, New York (1964). Mann, T. and U. Parsons: Effect of testicular hormone on the formation of seminal fructose. Nature, 160, 294 (1947). Price, D. and H.G. Williams-Ashman: The accessory reproductive glands of mammals. In Young, W. C. and Corner, G.W. (Eds.). Sex and Internal secretions. The Williams and Wilkins Co., Baltimore (1961), p. 366-448. Reddy, P.R.K. and J.M. Rao: Reversible antifertility action of testosterone propionate in human males. Contraception 5, 295-301 (1972). Roe, J.H. and C.A. Kuether: Determination of ascorbic acid in whole blood and urine. T.. Biol. Chem. 147, 399-407 (1943). Tyler, E.T.: Seminal fructose studies in infertility. Fertil. Steril. 6, 247-258 (1955). Warren, L., Thiobarbituric acid assay of sialic acids. J. Biol. Chem. 234, 1971-1975 (1959).
Acknowledgement: The authors wish t o thank Dr. R.K. Sanyal for his interest in this study. Skillful laboratory assistance of Shri H.C. Pandey is acknowledged.
Address: Dr. Somnath ROY, Biomedical Division, National Institute of Family Planning, D-5, Green Park, New Delhi-110016, India.
andrologia 7, Heft 3 (1975)
