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目的探讨载脂蛋白E(apolipoprotein E,APOE)在星形胶质细胞缺氧性损伤中的作用及相关机制。方法①原代培养APOE基因敲除鼠、APOE基因野生鼠的星形胶质细胞,并分别予以缺氧6 h,流式细胞仪分别检测两组细胞早期凋亡率情况;②将培养成熟的APOE基因野生鼠的星形胶质细胞分为常氧组、缺氧组和干预组(缺氧+ERK抑制剂),分别予以缺氧组和干预组缺氧6 h,干预组在缺氧之前向培养基中加入ERK信号通路抑制剂U0126,Western blot法分别检测检测3组细胞APOE蛋白表达变化情况。结果①缺氧6 h后,APOE基因敲除鼠组星形胶质细胞早期凋亡率[(5.67±0.35)%]明显高于APOE基因野生鼠组[(2.77±0.24)%](P<0.05);②与常氧组、干预组相比较,缺氧组星形胶质细胞表达的APOE蛋白明显增加(P<0.05)。结论 APOE对星形胶质细胞缺氧性损伤具有保护作用,缺氧后星形胶质细胞APOE蛋白水平上调与ERK信号通路有关。
Objective To investigate the role of apolipoprotein E (APOE) in hypoxic injury of astrocytes and its related mechanisms. Methods ① Primary cultured APOE knockout mice and APOE wild type astrocytes were exposed to hypoxia for 6 hours respectively. Flow cytometry was used to detect the early apoptotic rate in both groups. ② The mature APOE gene wild-type astrocytes were divided into normoxia group, hypoxia group and intervention group (hypoxia + ERK inhibitor), hypoxia group and intervention group were exposed to hypoxia for 6 h, ERK signaling pathway inhibitor U0126 was added into the culture medium, and the changes of APOE protein expression in the three groups of cells were detected by Western blot. Results ① After 6 h of hypoxia, the early apoptotic rate of astrocytes in APOE knockout mice was significantly higher than that in APOE transgenic mice [(5.67 ± 0.35)% vs (2.77 ± 0.24)%, P < 0.05). ② Compared with normoxia group and intervention group, the APOE protein expression in astrocytes of hypoxia group was significantly increased (P <0.05). Conclusions APOE has a protective effect on hypoxic injury of astrocytes. The up-regulation of APOE protein in astrocytes after hypoxia is related to ERK signaling pathway.