April Journal-2009.p65 - Association of Biotechnology and Pharmacy

Current Trends in Biotechnology and Pharmacy
Vol. 3 (2) 138-148, April 2009. ISSN 0973-8916
contrast, in the withaferin A treated group, the
number of cells were drastically decreased (Fig.
3A). This implied that withaferin A inhibit tumor
cell growth in vivo. The volume of ascites was
also measured using ascites from EAT bearing
as well as withaferin A treated EAT bearing
animals. The results indicated that withaferin A
treatment reduces the secretion of ascites fluid
(Fig. 3B). It is indicative from this data that
withaferin A is probably capable of inhibiting the
proliferation of tumor cell growth in vivo.
143
Withaferin A inhibits angiogenesis in vivo
Sprouting of new blood vessels is evident
in the inner peritoneal lining of EAT bearing mice.
The peritoneal lining of EAT bearing animals and
withaferin A treated mice was inspected for the
effect on tumor-induced peritoneal
neovascularization. EAT bearing mice treated with
withaferin A showed decreased peritoneal
angiogenesis when compared to the extent of
peritoneal angiogenesis in untreated EAT bearing
mice (Fig. 4A). Histopathological staining of
peritoneum sections from the EAT bearing group
appeared well vascularized. In contrast withaferin
A treated peritoneum sections were characterized
by a pronounced decrease in micro vessel density
and the caliber of detectable vascular channels.
While tumor bearing peritoneum sections showed
17 ± 1.2 blood vessels, withaferin A treated
peritoneum showed 6.8 ± 1.3 blood vessels (Fig.
4B).
Fig. 3: Effect of withaferin A on EAT cell number and
ascites volume in vivo
EAT cells (5X10 6 cells/animal, i.p.) were injected into
mice. After 6 days of tumor transplantation, withaferin A
(7mg/kg/animal) was injected on days 7th, 9th, 11th and
13th. The animals were sacrificed on days 8th, 10th, 12th
and 14th. EAT cells were collected along with ascites fluid.
Cells were counted in haemocytometer (A) and ascites
volume was measured (B). At least five mice were used in
each group and the results obtained are an average of three
individual experiments and means of ± S.E.M.
Fig. 4: Withaferin A inhibits angiogenesis and
microvessel density
A) Inhibition of peritoneal angiogenesis.
EAT bearing mice were treated with and without
withaferin A for four doses (7mg/kg/animal). The mice
were sacrificed and the peritoneal lining was observed
for extent of neovascularization. We presented
representative photograph of peritoneum.
B) Reduction in microvessel density (MVD)
The peritoneum of control as well as withaferin A
treated EAT bearing mice was embedded in paraffin
and 5ìm sections were taken using microtome. The
sections were stained with hematoxyline and eosine
and observed for microvessel density. Arrows indicate
the microvessels.
Prasanna et al