Protective Mechanism of Alisol A 24-Acetate on Brain Microvascular Endothelial Cells in Mice with Cerebral Ischemia-Reperfusion Injury by TRPM2/Akt/GSK3β Pathway

ObjectiveTo explore the mechanism of Alisol A 24-acetate (24A) on improving brain microvascular endothelial cells (BMECs) injury after cerebral ischemia reperfusion in mice.MethodsA total of 45 10-week-old male C57BL/6 mice were randomly divided into sham surgery group, model group and 24A group, with 15 cases in each group. The model group and the 24A group were treated with 20 min bilateral common carotid artery ligation followed by reperfusion to establish a cerebral ischemia reperfusion injury (CI/RI) model, but the common carotid artery and vagus nerve were separated without ligation in the sham surgery group. The 24A group was given 24A through intragastric administration with the dosage of 30 mg/(kg·d), and the sham surgery group and the model group were given equal volume of normal saline through intragastric administration. All three groups were given intragastric administration once a day, for seven consecutive days. Modified neurological severity score (mNSS) was used to evaluate neurological deficit; novel object recognition test (NORT) and Morris water maze (MWM) were used to evaluate the cognitive function; immunofluorescence was used to detect the expression of CD31 in cortex; Western blot was used to detect zonula occludens-1 (ZO-1), Occludin, Claudin-5, phosphorylated nuclear factor kappa-B (p-NF-κB), interleukin-1β (IL-1β) and tumor necrosis factor-α (TNF-α), transient receptor potential melastatin-2 (TRPM2), phosphorylated protein kinase B (p-Akt), phosphorylated glycogen synthetase kinase-3β (p-GSK3β) protein expression levels.Results(1) Neurological function: compared with the sham surgery group, mNSS in the model group significantly increased at the 1st, 3rd, 5th and 7th day (P