Evaluation of contraceptive properties of medicinal plant dioscorea bulbifear by in vitro sperm function and in vivo fertilization assay techniques
Date
2020
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Abstract
Dioscorea bulbifera is one of the medicinal plants which have shown to possess
various medicinal properties like anti- tumor, anti - inflammatory, anti- arthritis, anti-
fertility etc. D. bulbifera causes antifertility effect by suppressing ovulation and
implantation of embryos in females. The main objective of the present study is to
assess a direct contraceptive effect of Dioscorea bulbifera bulbil and tuber extracts
by investigating in vitro sperm function assays and in vivo fertilization (IVF)
techniques. Spectrometric analysis exhibited absorbance peak at 250 nm in both
extracts. The tuber extract showed distinct shoulder peaks at 280 nm and 340 nm
while such peaks were in conspicuous in the bulbil extract. Mouse epididymal
spermatozoa were incubated in capacitation medium with various concentrations of
extract. Spermatozoa were labeled with FITC peanut agglutinin (FPNA) or Coomassie
to study extract induced acrosome reaction. The treated spermatozoa were stained
with trypan blue or incubated with hypoosmotic medium to study viability and
membrane damage. Both extracts caused significant acrosome exocytosis at very
low concentrations. Comparatively, tuber extract resulted similar rate of acrosome
loss as bulbil extract. D. bulbifera tuber extract also caused membrane damage,
evaluated by trypan blue staining method and hypoosmotic swelling test. Viability
loss was remarkably lesser than acrosome exocytosis. SDS PAGE was performed for
protein profile analysis between the treated sperm sample and untreated control,
from which we observed no remarkable change on sperm protein bands in the
treated sperm cells. Finally, by performing in vivo fertilization test we have observed
that the extract treated female mice had reduced litter size. These results suggest
that D. bulbifera extract might effectively inhibit fertilization. The present research
will help in characterizing potential contraceptive effect of Dioscorea bulbifera and
in determining whether it could be used as a targeted and topical, contraceptive.
Keywords: Dioscorea bulbifera, anti- tumor, anti- inflammatory, anti-arthritis, anti-
fertility, in vivo fertilization, FITC peanut agglutinin, acrosome exocytosis,
contraceptive
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Keywords
Acrosome exocytosis, Dioscorea bulbifera