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Role of methandienone in haemopoiesis
Life Sciences Vol . 3, pp . 749-754, 1984. Pergamon Press, Inc. Printed in the United States
ROLE OF METHANDIENONE IN HAEMOPOIESLS A. K. Sanyal, C. R. Hanerjea and P. K. Das Department of Pharmacology, College of Medical Sciences Hauaras Hindu University, Varanasi-5, India (Received 14 May 1984; in final form 5 June 1984)
SYNTHETIC anabolic steroids with minimal of androgenic acrivity have recently been reported to have erythropoietic effect.
A favourable effect of nandro-
lone-phenyl propionate on a number of anaemic patients who did not respond to usual haemarinics has been reported.l Hooij and Kuypers 2 on the basis of their earlier observarion of "hyperhaemic" effect of nandrolone-phenyl propionate in two patients, conducted experimental studies with the steroid in normal albino rats.
They reported an increase in red cell count, haemoglobin percentage and
reticulocyte count in peripheral blood following treatment of these rats with nandrolone-phenyl propionate .
Aschkenasy3 has mentioned that 17-ethyl-l9-
nortestosterone is endowed with a certain degree of erythropoieric effect in female rats.
Encouraged by these reports, studies were conducted to find out
the effect of another anabolic steroid - methandienone (17-a-methyl-l7-ß-hydroxy androsta-1, 4-dien-3-one) on the normal and sub-normal haemopoiesis in albino rats and the results are reported here. Materials and Methods Experiments were conducted on adult albino rats of either sex weighing between 125 and 180 g.
The rats were divided into 8 groups of 8 rats each and
were treated with methandienone and/or chloramphenicol (details in Table 1 under Results) . During the whole course of experimentation and for at least one month prior to it all the animals were kept on the same diet.
Total leuco-
cyte and erythrocyte counts were done in each rat twice before drug administration and thereafter, at weekly intervals unril the animals were sacrificed after the experimental period.
Bone marrow smears of femur were also pre-
pared and examined immediately after the animals were sacrificed. 749
750
METHANDIENONE IN HAEMOPOIESIS
Vol . 3, No. 7
Results Methandienone (MD) in the dose of 5 mg/kg had no effect on erythrocyte and leucocyte counts while in the dose of 50 mg/kg it produced an insignificant increase in these counts .
Chloramphenicol (CP) in the doses of 100 and 200
mg/kg produced significant decrease in the erythrocyte count while marked leucopenia was observed with the higher dose only. MD 5 mg/kg had no effect
on the CP-induced anaemia while it significantly reduced the anaemia in 50 mg/ kg dose. MD in either of the doses completely antagonized the leucopenic action of CP (Table 1) . The effects of MD and CP alone and in combination on rat bone marrow are given in Table 2 . As the effects with both the dosage schemes with these drugs were qualitatively similar these have been grouped together . MD pro duced a general increase in the cellularity of the bone marrow especially of the erythroid cells as compared to leucocytic series of cells .
A compar. icon of
relative proportion of different cells showed that there was a greater preponderance of immature cells of the erythroid and granulocytic series (proerythroblasts, early and intermediate normoblasts ; myelocytes and metamyelocytes) as compared to the maturer cells .
There was, however, no significant
change in the lymphocytes and plasma cells .
CP produced generalized diminu-
tion of marrow cellularity . . However, at places there was also evidence of the presence of hypercellular foci in a few rats . of erythroid as well as leucocytic cells.
There was a generalized decrease
There was a marked reduction of
immature cells (proerythroblasts and early normoblasts) as compared to the maturer cells . control marrow. cells.
The number of metamyelocytes was also less compared to There was a detectable increase in lymphocytes and plasma
Haematogones and reticulum cells were also seen in the marrow smears
of CP treated rats. Combination of MD and CP showed a varied picture .
In a few rate, the
bone marrow picture simulated that of MD alone (hyperpla,stic erythroid reaction), in some it resembled those which had received only CP (hypoplastic marrow), while in others it simulated the normal bone marrow (normoplastic) . Incidental observation of superficial gastric ulcer was made in 5 out of
30 rats who received MD either alone or in combination with CP .
These ulcers
were accompanied with a fair degree of intragastric antemortem haemorrhage .
12
25
?5
3, MD 50 mg .
4 .. CP 100 mg .
5 . CP+100 mg.
5. 67+0, _ 68
7. 33+0 _ . 39
12
* MD- Administered orally
50 mg .
7,02±0,67
12
7, CP 200 mg. 8, CP 200 mg.
MD
7,35±0,55
12 -1 .8? -0,52
-0,40
-1. 66
-1,87
+0,35
-O,I2
-0 .20
10,81±1,34 11,51±1,34
8.97±1,02
9. 63+0, _ 98
10 .72+1,12
8.65±1,67
11 .78±1 .51
9,?5±1,34
Leuco te 7x~ia1~
** CP- Administered subcutaneously,
6,50t0,53
5.48~0 .62
6.55+0,38
5.18±0,56
7.05±0,49
6.95~0 .51
7.50±0,52
7,02±0,45
6.88±0 .52
7.15±0,71
7 .14±0, .61
7 .08±0.48
Er throcyte count in mill ions summ. Initial Terminal D fference
6, CP 100 mg. + MD 50 mg .
5 mg .
25
5 mg.
2 . MD
MD
25
No . of days .
1 . Control Saline 10 ml/kg .
Drug and Dose/day
10 .?5±1 .39
3.30±2 .82
9.11±1,23
10. 54+1 _ .21
9,88±1.42
10,52±1,08
10,57±1.22
10,51±1.19
Termina
-7,51 -0,76
+0.14
+0. 91
-0,84
+1.87
-1.21
+0,76
The Effect of Methandienone (MD) * and Chloramphenicol (CP) ** on Total Erythrocyte and Leucocyte Counts Expressed as Mean + Standard deviation.
TABLE 1
r
Ô ~d O
x
z
z o
ea
.-.
C 0
+
+ ?
6 . plasma cells
7 . Other cells
(+)
+++
++
+ +
+ +++
- -
+ + to ++
- -,
CP
+ to ++
+ to ++
+ to ++} + to ++
+ to +++ ++, +++
?
+ to +++ +, +++
+, N, -
MD + CP
PE : Proerythroblasts ; EN: Early Normoblasts ; IN : Intermediate Normoblasts ; LN : Late PIormoblasts ; M : Myelocytes ; MM : Meta Myelocytes . Plus and minus signs indicate relative proportion of different cells, ?s Occasional or Rare .
(Recticulum cells) Haematogones Haematogones & & iReticulum cells) (Reticulum cells)
+
+
+++ ++
5 . Lymphocytes
MM Mature
4 . Granulocytic series
++ +
++++ ++
++ ++
IN LN
+
?
3. Erythroid series pE
++++ ++ to ++++
++
MD
++ +++
N
Normal
2 . Predominant cell Erythroid Leucocytic
1 . Cellularity
Findings
TABLE 2 Effect of Methandienone (MD) and Chloramphenicol (CP) on Rags Bone Marrow
x0
ea
c
H
w N
Vol . 3, No . 7
METHANDIENONE IN HAEMOPOIESIS
753
Discussion The result of the present investigations showed that MD (5 and 50 mg/kg) administered to normal rats did not significantly affect the circulating erythrocyte and leucocyte counts .
An insignificant rise in the erythrocyte count was,
however, observed with the higher dose (50 mg/kg) of MD .
This observation,
therefore, is not in consonance with the findings of Hooij and Kuypers 2 who had reported significant increase in the haemoglobin and erythrocyte count in normal albino rats with nandrolonephenyl propionate in 2 . 5 mg, 5 mg, and 50 mg/kg dose.
They have not reported, however, the effect on the leucocyte count.
In the present series though there was no significant change in the circulating blood cells counts of normal albino rats, the bone marrow gave clear indication of hyperpla.sia with erythroid and leucocytic (myeloid type) reaction . Furthermore, MD in doses of 50 mg/kg was found to decrease the anaemic and leucopenic effects of CP.
The bone marrow of those rats treated
with both CP and MD usually gave a picture of either hyper- or normo-cellu larity with quite a few prcerythroblasts and early normobla.sts .
In a few rats,
however, the hypocellularity produced by CP could not be reversed by MD. These results indicate that MD not only causes hypercellularity of normal bone marrow but may also reverse the marrow picture in hypopla .etic conditions. The effect of androgens on bone marrow function is now well known. Polyglobulism can be provoked in guinea-pigs 4 and has also been seen in human beings 5 following treatment with male hormones.
Cases of aplastic anaemia
have also been treated successfully with androgens alone or in combination with prednisone. 6-9 However, the side effects of these agents are well known and may necessitate discontinuation of therapy . results may be unpredictable .
Moreover, in some cases the
The present studies provide some evidence that
primary anabolic steroids with minimal of adrogenic actions might also be clinically tried and may turn out to be better than hitherto used corticosteroids and androgens for the treatment of hypoplastic and apla.stic marrow cases . Summar~r The effect of methandienone (MD) has been studied on erythrocytic and leucocytic counts and bone marrow pictures of normal albino rats and in rats treated with chloramphenicol (CP) .
There was no significant increase in the
754
METHANDIENONE IN HAEMOPOIESIS
circulating erythrocyte counts with MD alone . showed hypercellularity with MD alone .
Vol . 3, No.7
The bone marrow, however,
CP produced decrease in erythrocyte
and leucocyte count in peripheral blood and hypocellularity of bone marrow . MD reduced the chloramphenicol induced anaemia, and leucopenia. and in most of the animals reversed the hypocellular (hypoplastic) marrow to hyper- or normo-plastic marrow .
The significance of the observations has been discus-
sed . Acknowledgement -- The authors thank M/S Ciba of India Limited for the generous supply of methandienone (Dianabol) used in this study. References 1.
J. K. MARTINS, Curr . Ther. Res . 3, 512 (1961) .
2.
J. BOOLT and J. KUYPERS, Acta Physiol . Pharmacol . Neerlandica 11, 12 (1962) . -
3.
A. ASCHKENASY, Therapie (Fr. ) 14, 332 (1959) .
4.
G. A. OVERBEEK, Ned . T. Geneesk . 90, i66 (1946) .
5.
B. J. KENNEDY and A. S. GILBERTSEN, New Eng. J. Med . 256, 719 (1957) .
6.
N. T. SHAHTDI and L. K. DIAMOND, A. M. A. J. Dias. Child . 98, 293 (1959) . -
7.
N. T. SHAHIDI and L. K. DIAMOND, New Eng . J. Med. 264, 953 (1961) .
8.
K. THOMAS, Klin. Wschr . 39, 55 (1961) .
9.
J. A. BELLANTI and D. PINKEL, J. Amer . Med. Ass . 178, 70 (1961) .
ROLE OF METHANDIENONE IN HAEMOPOIESLS A. K. Sanyal, C. R. Hanerjea and P. K. Das Department of Pharmacology, College of Medical Sciences Hauaras Hindu University, Varanasi-5, India (Received 14 May 1984; in final form 5 June 1984)
SYNTHETIC anabolic steroids with minimal of androgenic acrivity have recently been reported to have erythropoietic effect.
A favourable effect of nandro-
lone-phenyl propionate on a number of anaemic patients who did not respond to usual haemarinics has been reported.l Hooij and Kuypers 2 on the basis of their earlier observarion of "hyperhaemic" effect of nandrolone-phenyl propionate in two patients, conducted experimental studies with the steroid in normal albino rats.
They reported an increase in red cell count, haemoglobin percentage and
reticulocyte count in peripheral blood following treatment of these rats with nandrolone-phenyl propionate .
Aschkenasy3 has mentioned that 17-ethyl-l9-
nortestosterone is endowed with a certain degree of erythropoieric effect in female rats.
Encouraged by these reports, studies were conducted to find out
the effect of another anabolic steroid - methandienone (17-a-methyl-l7-ß-hydroxy androsta-1, 4-dien-3-one) on the normal and sub-normal haemopoiesis in albino rats and the results are reported here. Materials and Methods Experiments were conducted on adult albino rats of either sex weighing between 125 and 180 g.
The rats were divided into 8 groups of 8 rats each and
were treated with methandienone and/or chloramphenicol (details in Table 1 under Results) . During the whole course of experimentation and for at least one month prior to it all the animals were kept on the same diet.
Total leuco-
cyte and erythrocyte counts were done in each rat twice before drug administration and thereafter, at weekly intervals unril the animals were sacrificed after the experimental period.
Bone marrow smears of femur were also pre-
pared and examined immediately after the animals were sacrificed. 749
750
METHANDIENONE IN HAEMOPOIESIS
Vol . 3, No. 7
Results Methandienone (MD) in the dose of 5 mg/kg had no effect on erythrocyte and leucocyte counts while in the dose of 50 mg/kg it produced an insignificant increase in these counts .
Chloramphenicol (CP) in the doses of 100 and 200
mg/kg produced significant decrease in the erythrocyte count while marked leucopenia was observed with the higher dose only. MD 5 mg/kg had no effect
on the CP-induced anaemia while it significantly reduced the anaemia in 50 mg/ kg dose. MD in either of the doses completely antagonized the leucopenic action of CP (Table 1) . The effects of MD and CP alone and in combination on rat bone marrow are given in Table 2 . As the effects with both the dosage schemes with these drugs were qualitatively similar these have been grouped together . MD pro duced a general increase in the cellularity of the bone marrow especially of the erythroid cells as compared to leucocytic series of cells .
A compar. icon of
relative proportion of different cells showed that there was a greater preponderance of immature cells of the erythroid and granulocytic series (proerythroblasts, early and intermediate normoblasts ; myelocytes and metamyelocytes) as compared to the maturer cells .
There was, however, no significant
change in the lymphocytes and plasma cells .
CP produced generalized diminu-
tion of marrow cellularity . . However, at places there was also evidence of the presence of hypercellular foci in a few rats . of erythroid as well as leucocytic cells.
There was a generalized decrease
There was a marked reduction of
immature cells (proerythroblasts and early normoblasts) as compared to the maturer cells . control marrow. cells.
The number of metamyelocytes was also less compared to There was a detectable increase in lymphocytes and plasma
Haematogones and reticulum cells were also seen in the marrow smears
of CP treated rats. Combination of MD and CP showed a varied picture .
In a few rate, the
bone marrow picture simulated that of MD alone (hyperpla,stic erythroid reaction), in some it resembled those which had received only CP (hypoplastic marrow), while in others it simulated the normal bone marrow (normoplastic) . Incidental observation of superficial gastric ulcer was made in 5 out of
30 rats who received MD either alone or in combination with CP .
These ulcers
were accompanied with a fair degree of intragastric antemortem haemorrhage .
12
25
?5
3, MD 50 mg .
4 .. CP 100 mg .
5 . CP+100 mg.
5. 67+0, _ 68
7. 33+0 _ . 39
12
* MD- Administered orally
50 mg .
7,02±0,67
12
7, CP 200 mg. 8, CP 200 mg.
MD
7,35±0,55
12 -1 .8? -0,52
-0,40
-1. 66
-1,87
+0,35
-O,I2
-0 .20
10,81±1,34 11,51±1,34
8.97±1,02
9. 63+0, _ 98
10 .72+1,12
8.65±1,67
11 .78±1 .51
9,?5±1,34
Leuco te 7x~ia1~
** CP- Administered subcutaneously,
6,50t0,53
5.48~0 .62
6.55+0,38
5.18±0,56
7.05±0,49
6.95~0 .51
7.50±0,52
7,02±0,45
6.88±0 .52
7.15±0,71
7 .14±0, .61
7 .08±0.48
Er throcyte count in mill ions summ. Initial Terminal D fference
6, CP 100 mg. + MD 50 mg .
5 mg .
25
5 mg.
2 . MD
MD
25
No . of days .
1 . Control Saline 10 ml/kg .
Drug and Dose/day
10 .?5±1 .39
3.30±2 .82
9.11±1,23
10. 54+1 _ .21
9,88±1.42
10,52±1,08
10,57±1.22
10,51±1.19
Termina
-7,51 -0,76
+0.14
+0. 91
-0,84
+1.87
-1.21
+0,76
The Effect of Methandienone (MD) * and Chloramphenicol (CP) ** on Total Erythrocyte and Leucocyte Counts Expressed as Mean + Standard deviation.
TABLE 1
r
Ô ~d O
x
z
z o
ea
.-.
C 0
+
+ ?
6 . plasma cells
7 . Other cells
(+)
+++
++
+ +
+ +++
- -
+ + to ++
- -,
CP
+ to ++
+ to ++
+ to ++} + to ++
+ to +++ ++, +++
?
+ to +++ +, +++
+, N, -
MD + CP
PE : Proerythroblasts ; EN: Early Normoblasts ; IN : Intermediate Normoblasts ; LN : Late PIormoblasts ; M : Myelocytes ; MM : Meta Myelocytes . Plus and minus signs indicate relative proportion of different cells, ?s Occasional or Rare .
(Recticulum cells) Haematogones Haematogones & & iReticulum cells) (Reticulum cells)
+
+
+++ ++
5 . Lymphocytes
MM Mature
4 . Granulocytic series
++ +
++++ ++
++ ++
IN LN
+
?
3. Erythroid series pE
++++ ++ to ++++
++
MD
++ +++
N
Normal
2 . Predominant cell Erythroid Leucocytic
1 . Cellularity
Findings
TABLE 2 Effect of Methandienone (MD) and Chloramphenicol (CP) on Rags Bone Marrow
x0
ea
c
H
w N
Vol . 3, No . 7
METHANDIENONE IN HAEMOPOIESIS
753
Discussion The result of the present investigations showed that MD (5 and 50 mg/kg) administered to normal rats did not significantly affect the circulating erythrocyte and leucocyte counts .
An insignificant rise in the erythrocyte count was,
however, observed with the higher dose (50 mg/kg) of MD .
This observation,
therefore, is not in consonance with the findings of Hooij and Kuypers 2 who had reported significant increase in the haemoglobin and erythrocyte count in normal albino rats with nandrolonephenyl propionate in 2 . 5 mg, 5 mg, and 50 mg/kg dose.
They have not reported, however, the effect on the leucocyte count.
In the present series though there was no significant change in the circulating blood cells counts of normal albino rats, the bone marrow gave clear indication of hyperpla.sia with erythroid and leucocytic (myeloid type) reaction . Furthermore, MD in doses of 50 mg/kg was found to decrease the anaemic and leucopenic effects of CP.
The bone marrow of those rats treated
with both CP and MD usually gave a picture of either hyper- or normo-cellu larity with quite a few prcerythroblasts and early normobla.sts .
In a few rats,
however, the hypocellularity produced by CP could not be reversed by MD. These results indicate that MD not only causes hypercellularity of normal bone marrow but may also reverse the marrow picture in hypopla .etic conditions. The effect of androgens on bone marrow function is now well known. Polyglobulism can be provoked in guinea-pigs 4 and has also been seen in human beings 5 following treatment with male hormones.
Cases of aplastic anaemia
have also been treated successfully with androgens alone or in combination with prednisone. 6-9 However, the side effects of these agents are well known and may necessitate discontinuation of therapy . results may be unpredictable .
Moreover, in some cases the
The present studies provide some evidence that
primary anabolic steroids with minimal of adrogenic actions might also be clinically tried and may turn out to be better than hitherto used corticosteroids and androgens for the treatment of hypoplastic and apla.stic marrow cases . Summar~r The effect of methandienone (MD) has been studied on erythrocytic and leucocytic counts and bone marrow pictures of normal albino rats and in rats treated with chloramphenicol (CP) .
There was no significant increase in the
754
METHANDIENONE IN HAEMOPOIESIS
circulating erythrocyte counts with MD alone . showed hypercellularity with MD alone .
Vol . 3, No.7
The bone marrow, however,
CP produced decrease in erythrocyte
and leucocyte count in peripheral blood and hypocellularity of bone marrow . MD reduced the chloramphenicol induced anaemia, and leucopenia. and in most of the animals reversed the hypocellular (hypoplastic) marrow to hyper- or normo-plastic marrow .
The significance of the observations has been discus-
sed . Acknowledgement -- The authors thank M/S Ciba of India Limited for the generous supply of methandienone (Dianabol) used in this study. References 1.
J. K. MARTINS, Curr . Ther. Res . 3, 512 (1961) .
2.
J. BOOLT and J. KUYPERS, Acta Physiol . Pharmacol . Neerlandica 11, 12 (1962) . -
3.
A. ASCHKENASY, Therapie (Fr. ) 14, 332 (1959) .
4.
G. A. OVERBEEK, Ned . T. Geneesk . 90, i66 (1946) .
5.
B. J. KENNEDY and A. S. GILBERTSEN, New Eng. J. Med . 256, 719 (1957) .
6.
N. T. SHAHTDI and L. K. DIAMOND, A. M. A. J. Dias. Child . 98, 293 (1959) . -
7.
N. T. SHAHIDI and L. K. DIAMOND, New Eng . J. Med. 264, 953 (1961) .
8.
K. THOMAS, Klin. Wschr . 39, 55 (1961) .
9.
J. A. BELLANTI and D. PINKEL, J. Amer . Med. Ass . 178, 70 (1961) .