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.