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目的:观察缺氧/复氧条件下星形胶质细胞水通道蛋白(AQP)4和5表达的变化,探讨脑缺血再灌注后脑水肿与AQP4和AQP5的关系。方法:取新生24h内的SD大鼠皮质新鲜脑组织,进行原代和传代培养。以95%N2和5%CO2造成细胞缺氧,用倒置相差显微镜对细胞进行形态学观察,用锥虫蓝染色法测定缺氧及复氧后不同时间点星形胶质细胞的死亡数以反映星形胶质细胞的存活能力,应用细胞免疫化学技术测定星形胶质细胞缺氧及复氧后各个时间点AQP4、AQP5表达的变化。结果:缺氧4及8h后细胞形态变化不明显,随着复氧时间的延长出现细胞损伤的表现。与对照组比较,缺氧后4及8h有少量细胞死亡(P<0.05),随着复氧时间延长细胞死亡数亦逐渐增多(P<0.01);缺氧4及8h后,AQP4、AQP5阳性表达细胞数均减少(P<0.01),而复氧后AQP4、AQP5阳性表达细胞数逐渐升高并随时间延长呈增高趋势(P<0.01)。结论:星形胶质细胞对缺氧的耐受能力较强,但复氧时出现明显损伤;AQP4、AQP5表达的变化与缺血-再灌注损伤后脑水肿存在相关性。
AIM: To observe the changes of aquaporin 4 and 5 expression in astrocytes under hypoxia / reoxygenation conditions and to explore the relationship between cerebral edema and AQP4 and AQP5 after cerebral ischemia / reperfusion. Methods: Fresh cortex brain tissues of neonatal SD rats were cultured for primary and subculture. Cells were hypoxic exposed to 95% N2 and 5% CO2. Morphology was observed by inverted phase contrast microscope. Trypan blue staining was used to determine the number of astrocyte deaths at different time points after hypoxia and reoxygenation Astrocyte survival ability, the application of immunocytochemistry technology astrocyte hypoxia and reoxygenation at various time points AQP4, AQP5 expression changes. Results: After 4 and 8 h hypoxia, the morphological changes of cells were not obvious. The cell damage appeared with the prolongation of reoxygenation time. Compared with the control group, there was a small amount of cell death at 4 and 8 hours after hypoxia (P <0.05), and the number of cell death increased gradually with prolonged reoxygenation (P <0.01). After 4 and 8 hours of hypoxia, AQP4 and AQP5 were positive (P <0.01). After reoxygenation, the number of AQP4 and AQP5 positive cells gradually increased and increased with time (P <0.01). CONCLUSION: Astrocytes are well tolerated by hypoxia but have obvious damage during reoxygenation. The changes of AQP4 and AQP5 expression are correlated with cerebral edema after ischemia / reperfusion injury.