Virginia Angerclass

Beam [18]. Brain control group and i/r groups

Beam
walk test was employed to evaluate fore and hind limbs motor coordination. Each
animal was individually placed on a beam walk apparatus (Inco) made up of a
wooden bar 60 cm long and 1.5 cm wide, height 50 cm. The motor performance of
rat scored on a scale ranging from 1 to 4. This is a special test for animals
subjected to cerebral ischemia and reperfusion. For motor incoordination 1,number
of foot slip? 2, Number of falls? 1, Distance travelled along beam was studied
15.

Preparation of brain
homogenate

After
decapitation, ischemia/reperfusion groups the brain was removed and washed in
ice chilled 0.9% saline, and was kept on ice and subsequently blotted on filter
paper, then weighed and homogenized in cold phosphate buffer (0.1 M, pH 7.4) to
make a 10% w/v homogenate using a homogenizer. The homogenate was centrifuged
at 10,000 rpm for 20 min at 4ºC and supernatant was kept in deep refrigerator
at -80°C for analyzing biochemical estimations ( SOD, CAT,MDA) 16.

Infarct size measurement

The
brain was removed and frozen at -4°C for 5 min. Coronal slices were made at 1
mm to 2 mm and sections were immersed in 1% of 2,3,5-triphenyltetrazolium
chloride (TTC) at 37°C for 20 min. TTC is converted to red formazone pigment by
NAD and dehydrogenase present in living cells. Hence, viable cells were stained
deep red. The infarcted cells have lost the enzymes and thus remained
unstained. Whole brain slices were weighed. Infarcted unstained part was
dissected out weighted and expressed as % of total weight of brain 17.Histopathological
examination

At the end
of experimental procedures, rats were sacrificed and the brains were dissected
out. The brains were transferred to 10% formalin. Sections (<5 ?m thick) of the frontal forebrain were prepared and stained by hematoxylin and eosin for microscopical examination 18. Brain control group and i/r groups     S.N. Groups Neurodeficit score Falling time in sec Beam walk test 1 Shame Control 0 3.95±0.74 2.00±0.44 2 I/R group 3.66±0.21 1.13±0.24 3.00±0.36 2 ASE (200 mg/kg)      2.16±0.30** 3.46±0.39* 2.50±0.42 3 ASE (400 mg/kg) 1.33±2.95*** 4.08±0.47** 1.00±0.25** 4 DDE  (200 mg/kg) 2.16±0.47** 3.39±0.63* 2.00±0.25 5 DDE  (400 mg/kg) 2.00±0.36** 3.73±0.43** 1.46±0.12* 6 Combination C1   (Each 50mg/kg) 2.66±0.33* 3.5±0.50* 2.67±0.74 7 Combination C2 (Each 100mg/kg) 1.5±0.22*** 3.65±0.43** 1.00±0.25** 8 Vit E (150 g/kg p.o.)      1.66±0.33*** 4.02±0.45** 1.00±0.25**                           Results: MDA levels were significantly increased and SOD, CAT levels were significantly decreased in I/R of rats as compared to sham control group. The test drugs treated  treated groups, MDA levels were significantly reduced and SOD and CAT levels were increased significantly as the results are shown in table S.N. Groups CAT (U/mg of protien) SOD (U/mg of protien) MDA( nmole/gm of wet tissue   2 Shame Control 67.40±4.31 34.33±2.65 186.8±4.58 3 I/R group 34.40±2.60 15.50±1.78 366.6±16.85 4 ASE (200 mg/kg)      52±4.33* 29.00±3.86** 272.4±20.85**   5 ASE (400 mg/kg) 61.20±2.51*** 36.17±2.53*** 176.2±952*** 6 DDE  (200 mg/kg) 34.80±2.41 25.67±1.70* 281.0±22.72* 7 DDE  (400 mg/kg) 53.80±6.68** 27.83±1.62** 260.2±15.84** 8 Combination C1   (Each 50mg/kg) 36.80±2.47 20.33±1.49 328.0±33.05 9 Combination C2 (Each 100mg/kg) 55.60±3.02** 37.00±1.77*** 185.0±15.21*** 10 Vit E (10 g/kg)      59.40±2.67*** 37.17±2.70*** 162.0±5.17***