2.4 Infraction volume ratio and brain edema measurement
Excessive administration of pentobarbital sodium was employed for animal
euthanasia following ischemic conditions. The brains were sliced into
2mm coronal sections with even thickness. Subsequently, the sections
were subjected to staining using a 2% solution of 2,3,5-triphenyl
tetrazolium chloride (TTC) (Solarbio, Beijing, China) at a temperature
of 37°C for a duration of 20 minutes38. The stained
brain sections were fixed in 2% paraformaldehyde and imaged, followed
by analysis of the infarct volume (white) percentage using Image J
software from the National Institutes of Health, USA. In brief, the
infarct volume was calculated by multiplying the total areas of
infarction by the section thickness (2mm). The ratio of infarct volume
to total brain volume is indicative of cerebral infarction.
Brain edema was evaluated by measuring brain water content using the
dry-wet weight method39. In brief, following
reperfusion, mice were administered an overdose of anesthesia and their
brains were promptly harvested. The brains were then dissected along the
midsagittal plane to separate the ischemic hemisphere from the normal
hemisphere. Each hemisphere was immediately weighed, and subsequent
samples were dried in an oven at 100°C until a constant weight was
achieved to determine both wet weight and dry weight. The brain water
content was calculated as [(wet weight - dry weight)/wet weight] ×
100%.