EVALUATION OF NEUROPROTECTIVE
POTENTIAL OF TEST FORMULATIONS IN EXPERIMENTAL ANIMALS
1.0 TEST SYSTEM DETAILS:
Species : Rattus norvegicus (Rat)
Strain : Sprague
Dawley or Albino Wistar
Age : 6-10
weeks
Body Wight : 200-250 g
Sex : Male
No. of animals : 8 Animals in
each group
Total animals : 64+6 Extra
(Total = 70)
2.0 ALLOCATION OF GROUPS:
|
Groups |
Treatment |
Dose; ROA |
Animal
Numbers |
|
G1 |
Normal
Control |
Normal
saline or 0.25% Na-CMC |
8 |
|
G2 |
Sham (only Surgery) |
X mpk;
p.o. |
8 |
|
G3 |
BACCO
(Bilateral Common Carotid Artery Occlusion) Cerebral Ischemia |
X
mpk; p.o. |
8 |
|
G4 |
(Cerebral
Ischemia) Test
Formulation-1 |
X mpk; p.o. |
8 |
|
G5 |
(Cerebral
Ischemia) Test
Formulation-2 |
X
mpk; p.o. |
8 |
|
G6 |
(Cerebral
Ischemia) Test
Formulation-3 |
X
mpk; p.o. |
8 |
|
G7 |
(Cerebral
Ischemia) Test
Formulation-4 |
X
mpk; p.o. |
8 |
|
G8 |
(Cerebral
Ischemia) Test
Formulation-5 |
X
mpk; p.o. |
8 |
6 extra animals will be taken extra in
each study for the randomization purpose
3.0 METHOD:
· Healthy
animals will be selected, randomized based on body weight and divided into 8
different groups consisting of 8 animals each.
· Rats were anaesthetized with 75mg of Ketamine in Combination with 10mg
Xylazine ip.
· A midline incision
was made in the region between neck and sternum and trachea is exposed. Both the right and left common carotid arteries were located lateral to sternocleidomastoid, freed from surrounding tissues
and
vagus nerve separated.
· Cerebral ischemia was induced
by clamping both the arteries
with the help of aneurysm clips.
· After 10 min of cerebral
ischemia; the
clips
were removed
from the arteries to allow the reflow
of the blood
through carotid arteries.
· The incision was sutured back in layers with surgical suture.
While performing the surgical procedure; the body temperature was maintained at 37ÂşC by heated IR lamp or by any other
alternatives.
· Necropsy: The animals were deeply
anaesthetized with pentobarbitone.
Following decapitation,
the brains were taken
out and fixed in 10% formalin. Multiple, paraffin-embedded, coronal
sections
(5-Am thick)
were taken
from each brain (spanning
through striatum tocaudal hippocampus). Serial sections (spaced apart by 250 Am, 15 in total for each brain) were selected
for histopathological analysis of neuronal
damage.
4.0 END POINT PARAMETER(S):
·
Total protein
activity
·
Lipid Peroxidase
(LPO) activity
·
Reduced Glutathione (GSH) activity
·
Acetyl cholinesterase (AChE) activity
·
Measurement of infarction area
·
Brain Histopathology
5.0 REFERENCE(S):
5.1 Chandrashekhar
VM, Ranpariya VL. Neuroprotective activity of Matricaria Recutita Linn against
Global model of Ischemia in rats. J Ethnopharma. 2010;127:645-51.
5.2 Medhi B, Aggarwal R, Chakrabarti A. Neuroprotective effect of
pioglitazone on acute phase changes induced by partial global cerebral ischemia
in mice. Indian J Exp Biol Aug 2010;48:793-9.
5.3 Huisheng
Wu, Chaoliang Tang, Lydia Wai Tai, Weifeng Yao, Peipei Guo, Junmou Hong, et al,.
Research Article Flurbiprofen axetil attenuates cerebral ischemia/reperfusion
injury by reducing inflammation in a rat model of transient global cerebral
ischemia/reperfusion. Bioscience Reports (2018) 38 BSR20171562
https://doi.org/10.1042/BSR20171562
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