PROTECTIVE EFFECT OF Tribulus terrestris (NJERINJIL) IN CYCLOPHOSPHAMIDE TOXICITY ON REPRODUCTIVE SYSTEM OF MALE RATS
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Date
2018
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COLLEGE OF VETERINARY AND ANIMAL SCIENCES-MANNUTHY,THRISSUR
Abstract
The present study was aimed to evaluate the protective effect of Tribulus
terrestris (Njerinjil) in cyclophosphamide toxicity on reproductive system of male
rats.
The fruits of T. terrestris were procured locally, identified, dried in shade and
pulverized. Ethanolic extract was prepared by soxhlet extraction and qualitative
phytochemical analysis of the extract was performed to determine the active
components. Forty adult male Wistar rats were procured from Small Animal
Breeding Station, Mannuthy and divided into five groups of eight animals each.
Group I served as normal control, while group II, III, IV and V received
cyclophosphamide (CP) orally at the rate of 15 mg/ kg twice weekly for 30 days.
Groups III, IV and V were supplemented with ethanolic extract of T. terrestris fruit
daily at the dose rates of 100, 250 and 500 mg/kg respectively orally for 30 days
along with cyclophosphamide. Body weight of all the animals were recorded on days
0, 15 and 30.
On day 30, all the rats were sacrificed and dissected. After taking testes
weight, size and volume, the left testis and epididymis from each animal were used
for semen collection and for histopathological samples. The semen samples from
cauda epidiymis were used for estimation of semen parameters like mass activity,
sperm progressive motility, count, morphology, vital staining and acrosome integrity.
The other testis and epididymis were collected immediately on ice cooled bags for
the assessment of antioxidant assay of superoxide dismutase (SOD), reduced
glutathione (GSH) and lipid peroxidation (LPO) and functional marker enzyme levels
such as acid phosphatase (ACP), alkaline phosphatase (ALP), lactate dehydrogenase
(LDH) and sorbitol dehydrogenase (SDH). Representative samples of testes,
epididymis, liver, kidney and heart were fixed in 10 per cent neutral buffered
formalin and were stained with hematoxylin and eosin for histopathological
screening.
The preliminary phytochemical analysis of the ethanolic extract of T.
terrestris revealed the presence of various active principles like steroids, alkaloids,
glycosides, phenolic compounds, tannins, flavonoids, terpenes and saponins.
Administration of cyclophosphamide significantly reduced body weight,
relative testes weight, testes size, testes volume, mass activity, sperm motility, sperm
count, sperm viability and acrosome integrity, SOD, GSH, ACP and ALP. There was
also significant increase in per cent of abnormal sperms, LDH, SDH and LPO levels.
Histopathological studies in testis and epididymis confirmed reproductive toxicity in
the form of degeneration and loss of germinal epithelium within the seminiferous
tubules giving a washed out appearance. Liver showed centrilobular necrosis, fatty
degeneration and hepatocellular cytoplasmic vacuolation. Tubular epithelial
degeneration, interstitial haemorrhages, atrophy of glomerular tufts and formation of
casts within the lumen were observed in kidney. Sections of heart in GII animals
revealed myocardial degeneration, disruption and separation of cardiac muscle
fibres.
Treatment with ethanolic extract of T. terrestris at various doses significantly
increased body weight, relative testes weight, size and volume, mass activity, sperm
progressive motility, sperm count, acrosome integrity, SOD, GSH, ACP and ALP.
There was also a significant dose dependant decrease in per cent of abnormal sperms,
LDH, SDH and LPO levels with maximum protective effect at 500 mg/kg of the
extract. Histopathological examination showed a dose dependant regeneration and
restoration of seminiferous tubular epithelium in testes and ciliated lining epithelium
in epididymis. Reversion of hepatic and renal architecture to normal were observed
in T. terrestris extract supplemented groups. GIII, GIV and GV revealed normal cardiac
myocytes with centrally located nuclei having abundant cytoplasm outlined by
distinct and intact cell walls which was comparable to GI
.
Therefore, it could be concluded that ethanolic extract of T. terrestris has
protective effect and antioxidant potential in cyclophosphamide induced reproductive
toxicity in rats.
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