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目的:于大鼠海马原代培养的细胞模型中明确星形胶质细胞(Astrocytes,AS)在β-淀粉样蛋白(Am-yloid-β,Aβ)诱导的神经元凋亡中是否发挥作用,以及其发挥作用的主要方式。方法:建立4天龄Wistar大鼠海马原代混合培养体系(MIX-S)、纯化原代AS培养体系(AS-S)及纯化原代神经元培养体系(NE-S),用Aβ处理三种不同体系细胞,24 h后收集三种培养液(MNE-Aβ24 h、MAS-Aβ24 h、MMIX-Aβ24 h)及control组三种培养液(MNE-control、MAS-control、MMix-control)。在下一批NE-S和MIX-S培养细胞中,分别用不同培养液或Aβ处理24 h后应用TUNEL法检测细胞凋亡数量的变化。结果:Aβ处理后NE-S凋亡比例显著高于MIX-S,其中MIX-S凋亡与control组相近,MIX-S表现出较弱的Aβ毒性易感性。比较Aβ处理24 h后收集的培养液(MAβ24 h,Aβ前处理)与对照组培养液(Mcontrol)再加Aβ处理(Aβ后处理)培养细胞,发现Aβ前处理(MNE-Aβ24 h、MAS-Aβ24 h)比Aβ后处理(MNE-control、MAS-control再加Aβ)诱导NE-S凋亡的效果更加显著。而Aβ前、后处理对MIX-S组神经元诱导凋亡的效果类似,接近Control组凋亡比例。结论:AS参与保护Aβ诱导的海马神经元凋亡,其保护作用依赖于AS与神经元构建的MIX-S完整性而并不是由AS分泌入培养液的成分介导。单纯AS-S在Aβ刺激下不仅失去保护功能,且释放促凋亡物质。
OBJECTIVE: To determine whether astrocytes (AS) play a role in amyloid-β (Aβ) -induced neuronal apoptosis in primary cultured rat hippocampal cells, As well as its main ways to play a role. Methods: Primary culture system of hippocampus (MIX-S) was established in Wistar rats of 4 days old. Purified primary AS culture system (AS-S) and purified primary neuronal culture system (NE-S) (MNE-Aβ24 h, MAS-Aβ24 h, MMIX-Aβ24 h) and control group (MNE-control, MAS-control, MMix-control) were collected after 24 h. The next batch of NE-S and MIX-S cultured cells were treated with different culture medium or Aβ 24 h after the application of TUNEL method to detect changes in the number of apoptotic cells. Results: The ratio of apoptosis of NE-S after Aβ treatment was significantly higher than that of MIX-S. The apoptosis of MIX-S was similar to that of the control group, and MIX-S showed weak susceptibility to Aβ toxicity. Aβ pretreatment (MNE-Aβ24 h, MAS-PCR) was used to compare the effect of Aβ pretreatment (MAβ24 h, Aβ pretreatment) and control group Mcontrol (Aβ pretreatment) The effect of Aβ24h on NE-S apoptosis was more significant than that of Aβ (MNE-control, MAS-control plus Aβ). However, the effect of Aβ pretreatment and postconditioning on the apoptosis of neurons in MIX-S group was similar to that of control group. CONCLUSION: AS is involved in the protection of Aβ-induced apoptosis in hippocampal neurons. Its protective effect depends on the integrity of MIX-S constructed by AS and neurons, but not by the composition of AS secreted into the culture medium. AS-S alone not only loses its protective function but also pro-apoptotic substances under Aβ stimulation.