丁苯酞对缺氧/复氧诱导大鼠皮层神经元损伤的保护作用

Protective effect of dl-3n-butyphthalide against cortical neuron injury induced by hypoxia/reoxygenation in rats

目的观察丁苯酞对缺氧/复氧诱导的大鼠原代培养皮层神经元损伤的干预作用。方法原代培养Wistar大鼠皮层神经元,建立缺氧/复氧损伤的皮层神经元模型。实验分为5组:正常对照组、缺氧/复氧模型组、0.01μmol/L丁苯酞组、0.1μmol/L丁苯酞组、1.0μmol/L丁苯酞组。各组细胞进行相应的干预并孵育后,用CCK-8比色法观察神经元细胞活力,用激光共聚焦显微镜观察标记有荧光染料Fluo-3/AM的神经元细胞内游离钙离子浓度的变化。结果丁苯酞可抑制缺氧/复氧诱导引起的神经元细胞活力下降,且呈一定的浓度依赖;丁苯酞还可抑制细胞内游离钙离子浓度的升高,丁苯酞0.01,0.1,1.0μmol/L组与模型组相比差异有统计学意义(P<0.01)。结论丁苯酞可部分拮抗缺氧/复氧诱导的大鼠原代培养皮层神经元的钙超载,可能是丁苯酞对神经毒作用发挥保护作用的重要机制。

Objective To investigate the protective effect of dl-3n-butyphthalide on the injury induced by hypoxia/reoxygenation in primary cultured conical neurons of rats. Methods The hypoxia/reoxygenation injury model of cortical neurons was established in primary cultured Wistar rat cortical neurons. The model rats were randomized into 4 groups-model group and dl-3n-butyphthalide groups. The model rats in dl-3n-butyphthalide groups were given different doses of dl-3n-butyphthalide（0.01,0.1,1.0 ixmol/L） ,re- spectively. The vitality of neurons was tested by CCK-8 colorimetric assay, and the concentration changes of the free calcium ion labeled with Fluo-3/AM in neurons were detected by confocal laser scanning microscope. Results The decrease of neuronal vitality induced by hypoxia/reoxygenation was inhibited in a dose-dependent manner after giving d1-3n-butyphthalide. The increment of intracellular free calcium ion induced by hypoxia/reoxygenation was also inhibited after giving dl-3n-butyphthalide （ P 〈 0.01 ）. Conclusion DI-3n- butyphthalide can partially antagonize calcium overload induced by hypoxia/reoxygenation in primary cultured rat cortical neurons, which may be a significant mechanism of the protective effect of dl-3n-butyphthalide on neurotoxicity.