星形胶质细胞条件培养液对缺氧损伤神经元的影响

摘要

目的 分析星形胶质细胞(Ast)与腺苷对缺氧/复氧损伤神经元的保护作用,揭示腺苷对神经系统的保护机制.方法 体外培养SD大鼠大脑皮层Ast和海马神经元及大脑皮层神经元,纯化后给予神经元缺氧/复氧处理,收集复氧18 h后的Ast条件培养液(ACM)和腺苷预处理的Ast条件培养液(ACMa),然后用ACM、ACMa及ACM+腺苷(ACM+含100 μmol ·L-1腺苷的DMEM液)以1∶5的浓度培养缺氧损伤神经元;光镜观察神经元形态学变化,二甲氧唑黄比色法测定细胞活性.结果 ACMa组、ACM+腺苷组及ACM组的神经元缺氧损伤后的细胞形态较模型组得到明显改善,细胞活性较模型组也得到显著提高.不同条件培养液对缺氧/复氧后神经元活性的作用:ACMa＞ACM+腺苷＞ACM.结论 Ast条件培养液对缺氧/复氧损伤的神经元有重要的保护、修复作用,腺苷可通过Ast间接地保护和修复受损神经元.%Objective To analyse the protective effect of astrocytes and adenosine on the hypoxia / reoxygenation damaged neurons for further revealing the neuro-protective mechanisms of adenosine. Methods SD rat cerebral cortical astrocytes and astrocytes, hippocampal neurons were cultured in virto, neurons and hippocampal neurons were purified to establish models of hypoxia/reoxygenation injury. After reoxygenations for 18 liours,astrocyte condition medium( ACM) and adenosine-precondi-tioning astrocyte condition medium (ACMa) were collected. Then the damaged neurons, from models of hypoxia/reoxygenation injury were cultured in ACM,ACMa and ACM + adenosine (ACM mixed with DMEM medium containing 100 μmol · L-1 adenosine ). The concentration was 1:5. Light microscopy was used to observe the morphological changes of neurons and the neuro-nal activity was detected by XTT. Results Compared with model group, the cell morphology and neuronal activity improved obviously after neurons induced by hypoxia injury in ACMa group,ACM + adenosine group and ACM group. Their roles to the neuronal activity presumed to be;ACMa > ACM + adenosine > ACM. Conclusion Astrocyte-conditioned medium plays an important role in protecting and restoring damaged neurons. Adenosine can indirectly protect and repair damaged nerves through astrocytes,which plays key roles in hypoxia/reoxygenation injury.