2.3. Animals and experimental design
The Graduate Studies and the Ethical Committees of the College of
Pharmacy, University of Baghdad, approved the study protocol.
Wistar Albino experimental rats of both sexes aged six weeks with an
average weight of 150gm were utilized in this study; since the animals
were acquired and maintained in the College of Pharmacy Experimental
Animal House, University of Baghdad, Iraq; in addition, the experimental
rats were acclimatized for one week prior to starting the experiment;
the rats were housed under controlled conditions of a light/dark cycle
(12hours), temperature at (23±2°C) and humidity (50±5%); and had free
access to a standard commercial diet, which was purchased from the local
market, and tap water ad libitum .
The experimental animals (32 Rats) were randomly assigned into four
groups (n=8) as follows:
Group I : Each rat was orally administered vehicle only (5%
tween in DDW) via oral gavage for 14 consecutive days. Then a
single dose of normal saline (NaCl) (0.9%) was IP injected 1 hour after
the last vehicle administration on day 14. This group served as the
control group.
Group II : Each rat was orally administered DMF only
(15mg/kg/day) for 14 consecutive days.
Group III : Each rat was orally-administered vehicle (5% tween
in DDW) only via oral gavage for 14 consecutive days. Then a
single dose of DOX (90mg/kg) was IP injected 1hour after the last
vehicle administration on day 14. This group served as the model group.
Group IV : Each rat was orally administered DMF (15mg/kg/day)
for 14 consecutive days, then a single dose of DOX (90mg/kg) was IP
-injected 1 hour after the last DMF treatment on day 14.
Twenty-four hours after DOX injection (i.e., at day 15), rats were
anaesthetised using diethyl ether, blood samples were collected from the
Jugular vein in non-heparinized tubes and were left to clot at room
temperature, then centrifuged for 20 min at 4000 rpm to obtain serum and
stored at -20°C for biochemical analysis; the animals were sacrificed by
cervical dislocation, and the rats’ femoral bone marrow (BM) was
harvested and processed for genotoxicity evaluations (17).