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| Inhibition of Spermatogenesis of Albino Rats Through Feeding of Lantana camara Leaf Powder | ||||||||||||||||||||||||||||||
| Paper Id :
17469 Submission Date :
2022-12-04 Acceptance Date :
2022-12-21 Publication Date :
2022-12-25
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| Abstract |
Recently many plants have been reported for regulating fertility of male and female animals (Chaudhary, 1966) affecting the function of the reproductive organs (testes and ovaries respectively). Lantana camara (Family - Verbenaceae, commonly known as Kurri in Hindi and wild sage in English), is an exotic plant found abundantly in India. Fertility regulating activity of this plant was reported earlier (Singh et al., 2000), but its effect on male's spermatogenesis was not yet studied. The same is reported in this paper. Leaf powder of Lantana camara, as aqueous solution at doses of 20, 40 and 60 mg/kg were orally fed to three groups (dosewise) with a control group (vehicle treated) for 30 days. Five male rats were included in each group. On day 31st, all the rats were killed, weighed and dissected. Testes and epididymes were taken out, weighed and processed for histological examination. Weight of reproductive organs were significantly reduced at higher doses. The spermatogenesis was arrested in seminiferous tubules. Leydig's cells were atrophied. Epididymes were devoid of spermtozoa. It is concluded that this plant inhibit the spermatogenesis of male rats.
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| Keywords | Lantana camara, Phytomedicine, Herbal drugs, Animal reproduction, Medicinal plants, Fertility control. | |||||||||||||||||||||||||||||
| Introduction |
Lantana camara (Family : Verbenaceae – commonly known as 'Kurri') is an exotic plant came from America years ago. It is a fast growing perennial herbaceous plant found abundantly in each and every corner of India. It is never consumed as a fodder by domestic and wild animals. It grows under varying conditions of climate and soil including gravel and laterite and in low lying areas and on hills up to 1800 m. It is a draught resistant, light loving and tolerate moderate shades.
As far as human is concern, the leaf oil is used in treatment of itches of skin and antiseptic for wounds. The plant is considered vulnerary, diaphoretic, and antiseptic. It is useful for fistulae, pustules and tumours. A decoction of the plant is given in tetanus, rheumatism, malaria and atoxy of abdominal viscera. In Philippines, a decoction of fresh roots is used as gargle for toothache. Powdered leaves are applied to cuts, ulcers and swellings. A decoction of leaves and fruits is used as a lotion for wounds.
It is thought that the plant may cause adverse effect on reproductive potential of animals.
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| Objective of study | 1. To find out the effect of Lantana camara on male reproductive organ of study model.
2. To study the inhibition of spermatogenesis of male rats using leaf powder of Lantana camara |
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| Review of Literature | Earlier Singh et al. (2000) reported antifertility effect of leaves of
this plant on female albino rats but no such effect on male animals reported so
far. Plants affecting reproductive function (anti-spermatogenic and
anti-ovulatory etc.) have been already reported by Kirtikar and Basu, 1935;
Nadkarni and Nadkarni, 1954; de Laszlo and Henshaw, 1954; Casey, 1960;
Chaudhary, 1966; Malhi and Trivedi, 1972; Saxena, 1973; Farnsworth et al.,
1975; Kamboj and Dhawan, 1982; Singh, 1985; Gupta et al., 2005; Singh and Singh
2017; Singh & Purohit. 2019 and Singh et al., 2021. Authors have conducted
experimental study of leaf powder of Lantana camara in male albino rats
and the results are being reported in this paper. |
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| Main Text |
Observations A.
Effect on body and organ weight The
result are presented in Table 1. The rats of control group did not show much
change in the body weight. It is slightly increased after 30 days of
experimental period. Experimental male rats administered with leaf powder
of Lantana camara showed some reduction in te body weight. The
doses of leaf powder at 40 and 60 mg/kg/day for 30 days caused significant
reduction (P < 0.05) in the weight of testes and epididymes in comparison to
control rats. B. Effect
on testes Control
: The
testes of control male rats after 30 days showed normal histological features.
The seminiferous tubules were lined with germinal epithelium, whose cells by a
process of growth and division (Spermatogenesis) produced the spermatozoa.
There were a kind of specialized and large sized cells, the sertoli cells,
placed between the epithelial cells of seminiferous tubules. Leydig's cells
were present in the interstitium which secrete the male hormone, testesterone
largely responsible for the development of secondary sexual characters. Other
spermatogenic cells, spermatocytes, spermatids and spermatozoa are visible in
the normal sized seminiferous tubules (Fig. 1). Treated
with doses : Administration
of Lantana camara leaves powder as aqueous solution at 20
mg/kg dose for 30 days caused no significant histological changes in the
seminiferous tubules of testes. Various cell types including spermatozoa are
normal. The administration of 40 mg/kg dose for 30 days caused mild
degenerative changes in the testes. The spematogenesis was stopped at spermatid
stage in majority of seminiferous tubules. Germinal epithelium appeared normal.
The degenerated Leydig's cells were observed in the intertubular spaces
(interstitium). The administration of 60 mg/kg dose for 30 days caused severe
degenerative changes. The degeneration in the testes consisted of the damage of
the spermatocytes, spermatids and spermatozoa. The seminiferous tubules were
reduced in size, become deshaped and filled with debris of connective tissues.
The intermediate contained atrophied Leydig's cells. The vascularity was also
affected (Fig. 2). C. Effect
on Epididymes Control
: The
epididymes of control rats showed a normal histological structure. The
epithelium of caput and cauda parts were normal. The columnar epithelial cells
of caput were tall with basal nuclei (Fig. 3). The epithelium cauda was lined
with low cuboidal cells (Fig. 4). The lumen of caput was smaller than the cauda
ductules. The pronounced stereocilia were present in the caput than in the
cauda. The intertubular connective tissue with vascularity between the ductules
of caput and cauda were normal. Experiments have shown that when spermatozoa
passed through the epididymes, they undergone a physiological maturation. The
epididymes both caput and cauda were full of mature spermatozoa. Treated
with doses : Administration
of Lanatana camara leaves powder as aqueous solution of 20
mg/kg dose for 30 days caused no histological changes in both caputa and cauda
epididymes. The doses 40 and 60 mg/kg for 30 days caused marked changes in the
histology of both caput and cauda epididymes. The epithelium of caput was
slightly hypertrophied thereby showing the increased height of epithelial cells
and wide lumen. The nuclear dysplasia was observed greatly in the epithelial
cells of caput than in the cauda part. Sperm concentration became very less in
the lumen of both caput and cauda ductules. Stereocilia were severely affected
in both parts of epididymes (Fig. 5 and 6). Discussion The
leaf powder of L. camara as aqueous solution administration
orally to adult male albino rats caused marked inhibitory effects of
spermatogenesis at the doses of 40 and 60 mg/kg for 30 days. The dose 20 mg/kg
was ineffective. The effect on body weight was negligible but the higher doses
for 30 days reduced the weight of male reproductive organs i.e. testes and
epididymes significantly. Not only weight reduction was noticed in the
reproductive organs but, feeding of L. camara leaf powder,
caused histopathological changes in testicular elements. Leydig's cells were
mostly atrophied which are the main testosterone producing cells. The
Follicle Stimulating Hormone (F.S.H.) secretion is directly related to the
weight of testes. Heavier will be the testes more will be secretion of F.S.H.
Testosterone is an androgenic hormone essential for sexual development.
Chemically, it is a steroidal hormone. It is confirmed by studies of Dorfman et
al. (1963) that both non-steroidal (i.e. plant product etc.) and steroidal
agents inhibits pituitary gonadotrophins either acting directly on pituitary or
through the hypothalamo-hypophyseal axis. The reduced F.S.H. production causes
significant decrease in the weight of the testes and accessory reproductive
organs of male rats. Paul et al., (1953) had demonstrated the reduction of
weight of testes and accessory organs in absence or presence of spermatids and
spermatozoa in the testes of male animals. According to Nelson and Patanelli
(1965), the change in weight (reduction) of testes also corresponds to the
presence or absence of post meiotic cells in the testes. The physiology of male
reproductive (genital) organs are dependent on androgen hormone. Findings of
the present study are in agreement with the above stated studied of male
reproductive physiology with the following references. The aqueous leaf extract
of Vinca rosea caused significant histological changes in
testes and epididymes and the weight reduction also (Chinoy and Geetha Ranga,
1983). The results of anti-spermatogenic effects of leaf extracts of Aristolochia
indica in male rats by Pakrashi and Pakrashi (1977) may be mentioned
for the support of present studies. Administration of Ocimum sanctum L.
leaves by Kashinathan et al., 1972; Khanna et al., 1986 and Cannabis
sativa L. leaves by Dixit et al. (1974) to male rats and mice
respectively concerning with degenerative changes in the testes can be
corroborated. Ocimum sanctum caused adherence of
spermatological cells and regression of Leydig's cells. Cannabis sativa also
induced Leydig's cells regression and reduction in size and spermatocytes
population of seminiferous tubules in the testes of mice. Singh
(1985) reported degenerative changes at higher doses of Semicarpus
anacardium Linn. Spermatogenic cells were degenerated. In some of the
germ cells, the nuclei were swollen. Seminiferous tubules were deshaped and
distorted. Sperm concentration in the ductules of epididymes were greatly
reduced. Chaudhary et al. (1990) reported effects of leaf extracts of some plants
in male rats which caused antifertility in them. Administration of these plants
caused mass atrophy of the spermatogenic elements. The germ cells were left 1
to 2 layers only. Epididymal epithelium was regressed and the lumen was
devoid of spermatozoa. Table
1 : Effect of leaf powder of Lantana camara on body wt. (gms)
and wt. of Reproductive organs (mgs) of male albino rats administered for 30
days with doses of 20, 40 and 60 mg/kg/day. Five rats were used in each group.
Values are mean ± S.E.
*
p < 0.05
Fig.1. Section of testes of control group for 30 days showing normal histological features in seminiferous tubules. Sprmatogenesis with germinal epithelium and layers of spermatogonial cells, spermatocyte, spermatogonia, spermatids and spermatozoa towards lumen. Leydig's cells are normal in the intertubular spaces´250
Fig.2. Section of testes of treated group with Lantana camara at 40 and 60 ml/kg dose for 30 days caused severe degenerative changes, deshaped seminiferous tubules, damaged spermatogonia, lumen without spermatozoa and atrophied Leydig's cells in the intertubular spaces 250.
Fig.3. Section of epididymes (caput) of control group of male rats for 30
days showing normal structure of tall columnar cells in the epithelium of
ductules with spermatozoa in the lumen. Stereocilia are normal ´ 250.
Fig.4. Section of epididymes (cauda) of control group of male rats for 30
days showing normal structure of cuboidal cells in the epithelium of ductules
with spermatozoa in the lumen. Stereocilia are normal ´ 250.
Fig.5. Section of epididymes (caput) of treated group with Lantana camara for 40 and 60 mg/kg doses for 30 days caused vacuole formation in epithelial cells of ductules, distorted stereocilia and absence of spermatozoa in lumen ´ 250.
Fig.6. Section of epididymes (cauda) of treated group with Lanata camara for 40 and 60 mg/kg doses
for 30 days caused distortion of epithelial cells of deshaped ductules with
vacuoles also, damaged stereocilia and absence of spermatozoa in the lumen ´ 250. |
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| Methodology | 1. Albino Rats (Rattus rattus norvegicus)
Healthy, adult, colony-breed, Swiss albino male rats, three-four months old weighing between 150-200 gms were selected and acclimatized to the laboratory condition for seven days prior to commencement of experiment. The rats were kept in cages (60cm 45cm 45cm) under normal conditions of photoperiod and at room temperature. Four groups of male rats each containing five albino rats were made. First group of rats were served as control while three groups of rats were treated as experimental. They were maintained on balanced diet containing soaked gram and food pallet (Hindustan Lever's Limited) and pure hygienic drinking water was allowed ad libitum. All the rats were cared as per U.G.C. guidelines and supervised by the members of Animal Ethical Committee appointed by the than Principal of the college.
2. Doses and Administration
Fresh leaves of Lanatana camara were collected from Dehradun city barren lands, washed and dried in shade. They were crushed until change into fine powder. Three doses 20, 40 and 60 mg/kg body weight were prepared. Each dose was dissolved in distilled water until it changes into a homogenous mixture. The volume was adjusted in such a way that 01 ml of the solution containing 20 mg powder fed to second group of rats for 30 days. The rest of the doses 40 and 60 mg were administered in the 3rd and 4th group of rats respectively for 30 days. Gum-acacia powder as vehicle dissolved in water (05 mg/dose) was fed to control group (Ist) of rats for 30 days. The administration of doses were done orally with specially designed knobbed feeding needle fitted into a syringe.
3. Record of weight and Histology
The initial and final (on day 31st) body weight of the experimental as well as control rats for 30 days of treatment were recorded. The rats from all groups of treated and control were sacrificed after 24 hours of the last dose under light either anaesthesia and dissected quickly. Testes and epididymes were taken out, freed from adjoining tissues and blood. Weight of testes and epididymes were recorded in a semi-micro balance. The testes and epididymes were washed thoroughly in 10% of saline solution, fixed with Bouin's fluid (for two days), washed in tap water, dehydrated in alcoholic series and embedded in paraffin wax at 60C. The blocks of reproductive tissues were sectioned at 5 (micron) using Rotary Microtome and stained with haemotoxylin and eosin for histological study. The slides were observed under microscope at high power and photographed at different magnification.
4. Statistical Analysis
The body weight and organ weight (testes and epididymes) of both treated and control group of male rats were recorded in tabular form. The data were statistically analysed using student 't' test. The values were expressed as mean standard error (S.E.). The significance of weight between treated and control rats were taken at P < 0.05 as significant. |
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| Conclusion |
It is concluded that the powdered leaves of Lantana camara have potentiality to regulate the male fertility by causing histopathological changes in the reproductive organs and inhibition of spermatogenesis. |
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| Acknowledgement | Authors are thankful to Principal for providing Laboratory facilities and encouragement. Sincere thanks to U.G.C., New Delhi for financial support in the form of Minor Research Project No. 8-7(54) 87 (S.R.-II). | |||||||||||||||||||||||||||||
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21. Singh, S.P. and Singh, Shiv Pratap (2017). Anti-Spermatogenic effect of Solanum xanthocarpum in male Guinea pigs. Universitie's Jour. Phytochem. and Ayurv. Heights, Vol. II(23) : 58-61.
22. Singh, S.P., Pathania, P.C. and Bhatia, D.K. (2000). Effect of Lantana camara L. leaves on the ovarian and uterine histology, oestrus cycle and ovulation in female albino rats. Indian Jour. Env. Sci., 4(1) : 53-60.
23. Singh, S.P. and Purohit, M.K. (2019): Anti- Implantation effects of Abrus precatorious (seed) infemale albino rats. J.Env.Bio.Sci. 33(2):321-323.
24. Singh, S.P., Purohit, M.K. and Shruti Saxena, (2021): Effect of Cannabis sativa on spermatogenesis of male albino rats. Universities Journal Phytochem. & Ayurv. Heights. 11(31):34-40. |
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