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Effect of Sarcostemma acidum stem extract on spermatogenesis in male albino ratsPramod Kumar Verma1, Anita Sharma, Annu Mathur, Prachi Sharma, R.S. Gupta, S.C. Joshi, V.P. Dixit Reproductive Physiology Section, Department of Zoology, University of Rajasthan, Jaipur-302 004, India Asian
J Androl 2002
Mar;
4: 43-47 Keywords:
|
Treatment |
Initial
weight(g) |
Weight
after 2 weeks (g) |
Weight
after |
Weight
after 6 weeks (g) |
Weight
after 8 weeks (g) |
Control |
21814 |
23018 |
24520 |
25628 |
28516 |
S.acidum
stem 50 mg/kg BW/d |
20018 |
22312 |
24219 |
26021 |
28020 |
S.acidum
stem |
23021 |
24813 |
25714 |
29018 |
30525 |
Table 2. Effect of S. acidum stem extract on fertility of the male rats. n=10 for each group.
Treatment |
No
of females |
No
of females |
Fertility |
Control |
20 |
20 |
100 |
S.acidum
stem |
20 |
14 |
20 |
S.acidum
stem |
20 |
0 |
NIL |
Table 3. Effect of S. acidum stem extract on sperm density and motility and some biochemical markers in the rats. n=10 for each group, mean SEM, cP<0.01 vs control.
Treatment |
Sperm
motility |
Sperm
density |
Testes |
Testes
protein |
Testes |
Seminal
vesicle |
Epididymides |
Control |
71.9
0.7 |
56.8
1.5 |
4.8
0.3 |
188
3 |
3.8
0.2 |
4.5
0.2 |
248
3 |
S.
acidum stem |
35.6
2.9c |
14.4
1.3c |
6.2
0.2c |
144
11c |
3.1
0.1c |
2.5
0.1c |
202
6c |
S.
acidum stem |
40.7
2.8c |
5.8
1.0c |
7.1
0.5c |
144
6c |
2.4
0.3c |
3.0
0.1c |
149
8c |
3.2 Blood and serum analysis
Blood variables, i.e., RBC and WBC counts, hemoglobin, haematocrit, sugar and urea were within the normal range. Cholesterol and protein did not changed significantly in any of the treatment but phospholipids were decreased only at the 100 mg dose level (Table 4).
Table 4. Effect of S. acidum stem extract on some components in whole blood and blood serum of rats. n=10 for each group, mean SEM, aP>0.05, cP<0.01 vs control.
Treatment |
RBC
count |
WBC
count |
Hemoglobin |
Haematocrit |
Blood |
Blood |
Serum |
Serum |
Serum
protein |
Control |
4.7
+ 0.1 |
8540
81 |
13.4
0.3 |
44.6
4.4 |
86.0
1.6 |
35.6
2.5 |
89
5 |
86
6 |
13530
594 |
S.
acidum stem |
4.7
0.2a |
8317
78a |
11.9
0.4a |
40.1
4.4a |
76.9
6.3a |
42.0
3.3a |
81
4a |
81
4a |
13000
444 |
S.
acidum stem |
4.6
0.1a |
8300
100a |
12.6
0.2a |
43.6
0.8a |
75.5
8.9a |
40.5
3.0a |
69
6a |
55
4c |
11556
544 |
3.3 Biochemical findings
Glycogen and protein contents in the testes were decreased significantly (P< 0.01) after treatment with S. acidum stem in comparison to the control. Cholesterol level in testes was elevated by 29.2% and 47.3% at 50 and 100 mg dose levels, respectively. Seminal vesicular fructose and epididymal proteins were decreased significantly (P<0.01) after both doses (Table 3).
3.4 Histological observation
In the treated groups, there was a significant reduction in the number of primary and secondary spermatocytes and round spermatids, while the numbers of the spermatogonia and Sertoli cell remained unchanged. There was 49.6% and 40.7% reduction in the Leydig cell nuclear area at 50 and 100 mg doses, respectively. The number of mature Leydig cell was decreased significantly (P<0.01), whereas degenerating cell number was increased in comparison to control animals (Table 5, Figures 1-3).
Table 5. Effect of S. acidum stem extract on spermatogenesis and Leydig cells in the rats. n=10, mean SEM, bP>0.05, cP<0.01 vs control.
Treatment |
Sertoli |
Spermato- |
Preleptotenes |
Pachytene |
Secondary |
Round |
Mature |
Degenerating |
Leydig
cell |
Control |
3.20.4 |
5.90.6 |
19.92.1 |
24.62.6 |
44.83.4 |
32.21.6 |
63.10.5 |
36.93.3 |
28.01.2 |
S.
acidum stem |
3.30.2b |
3.40.5b |
6.90.3c |
19.30.1b |
25.91.7c |
18.52.1c |
45.81.3c |
54.20.3c |
14.10.8c |
S.
acidum stem |
2.70.1b |
5.70.4b |
12.40.7c |
14.31.0c |
28.41.7c |
12.61.2c |
37.92.2c |
62.12.1c |
16.60.7c |
Figure
1. Control rat testis showing normal spermatogenesis. 200
HE.
Figure 2. S. acidum stem
extract 50 mg/kg BW/d. Microphotograph of testis showing reduced number
of spermatogenic cells. 200
HE.
Figure 3. S. acidum stem
extract 100 mg/kg BW/d. Microphotograph of testis showing arrest of spermatogenesis
at spermatocyte level and degeneration of Leydig cells. 200
HE.
4 Discussion
Sarcostemma acidum stem suppressed sperm production, as evidenced by the reduction in the number of spermatogenic cells. Similar results were found by the administration of Colebrookia oppositifolia in rats[14] and Mentha arvensis in mice [15]. The principal cells of epididymis synthesize proteins which have important role for maturation of spermatozoa [16]. Alteration in secretion and function of these proteins caused incomplete maturation of spermatozoa with a decline in sperm motility. In the present study the epididymal protein was also decreased by S. acidum stem extract feeding. Low fructose concentration may be another cause of low sperm motility. Chinoy and Bhattacharya[17] reported reduced sperm motility after aluminium chloride adminimistration in mice with decreased seminal vesicular fructose, as the latter supplies energy for sperm motility. Reduced glycogen reflects decreased number of post-meiotic germ cells, which are thought to be the sites of glucose metabolism [18].
Cholesterol is involved in steroidogenesis in testes. It is most important precursor in synthesis of steroid hormones and its level is related to fertility of individuals [19]. Increased level of cholesterol may be due to decreased androgen production, which resulted in accumulation of cholesterol in testes, hence impaired spermatogenesis [20].
The impairment of Leydig cell function was evinced by its reduced nuclear area and lower number of mature Leydig cells. The number of mature Leydig cells has a direct bearing on spermatogenesis [21]. Deformation of Leydig cells further indicates the inefficiency of these cells to synthesize testosterone [22].
Reduction in number of spermatogenic cells may be due to insufficient amount of testosterone. Spermatogenesis is activated by testosterone which is synthesized by Leydig cells and act on Sertoli cells, and peritubular cells [23]. Similar results were reported by Gupta et al [21] in the Barleria prionitis root extract treated rats. The number of Sertoli cells and spermatogonia (stem cells) were not changed after the treatment, so 100% fertility could be retained after cessation of the treatment.
It has been observed that the blood and serum parameters were within the normal range, indicating non-toxicity of S. acidum stem on general body metabolism.
Our results reflect antispermatogenic/antiandrogenic effects of S. acidum stem in male albino rats, without affecting general body metabolism.
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Correspondence
to: Dr. Pramod Kumar Verma,
C/o Dr. L.M.Mathur, B-112, Sethi Colony, Jaipur-302004, India.
Tel: +91-141-611 114, 617 969
E-mail: pramod@cswri.raj.nic.in,
verma23@rediffmail.com
1 Present Address: Dr. Pramod Kumar Verma, Division of Physiology, Central
Sheep and Wool Research Institute, Avikanagar (via-Jaipur)-304501, Rajasthan,
India.
Received
2001-05-28 Accepted 2002-02-20