2.1 Mice and cerebral ischemia reperfusion model
One hundred and sixty C57BL/6J male mice aged 8-12 weeks with a body
weight of 20-30g were housed under controlled environmental conditions
with a consistent temperature of 22-25℃, relative humidity of 65%, and
12h light/dark cycle and in standard cages. All the mice were provided
with unrestricted access to food and water. All experimental protocols
and animal handling procedures in this study were conducted in
accordance with the National Institutes of Health (NIH) guidelines for
the use of experimental animals, and the study received approval from
the Institutional Animal Care and Use Committee of the Air Force Medical
University (No. IACUC-20220122).
Referred to in a prior investigation34, The middle cerebral artery
occlusion model was successfully established in our research. Cerebral
ischemia was induced by middle cerebral artery occlusion (MCAO). General
anesthesia was induced with 5% isoflurane (RWD, Shenzhen, China) in a
mixture of 70% nitrous oxide and 30% oxygen, and maintained with 2%
isoflurane delivered via facemask during spontaneous respiration. Rectal
temperature was closely monitored and maintained within the range of
36.5-37.5°C throughout the surgery until the animal regained
consciousness, with the aid of warming blankets and lamps. Briefly, a
midline incision was made in the jugular skin to expose the unilateral
common carotid artery (CCA), external carotid artery (ECA), and internal
carotid artery (ICA). Subsequently, the CCA and ECA were ligated while
the ICA was clipped using microvascular aneurysm clips. Following
arteriotomy in the ECA, a soft silicone-coated surgical nylon
monofilament suture (0.12 mm diameter; 3.0 cm length, RWD Life Science,
China) was gently inserted into the ICA via the ECA to occlude the
middle cerebral artery (MCA), approximately 12.0 mm distal to carotid
bifurcation. Reperfusion was allowed by removing filaments after a
2-hour period. Neurological deficit scores were evaluated at 24 and 48
hours post-MCAO, followed by decapitation to obtain the brains for
subsequent experiments. In the Sham group, mice underwent the same
procedure without the insertion of a nylon monofilament to occlude the
middle cerebral artery (MCA). Intraperitoneal administration of 0.4 mg/g
D-allose, which was soluble in normal saline, was given within 5 minutes
after reperfusion. The dosages utilized in this study were determined
based on our preliminary experiments and previous
research35.