氟桂利嗪阻滞小鼠皮质神经元胞浆钙离子内流的实验研究

Flunarizine inhibits the calcium ion influx in cultured mouse cortical neurons

目的 研究缺血状态下神经元胞浆内游离钙离子的变化及氟桂利嗪阻滞钙离子内流的作用。方法 用Flu 3 AM作为细胞内钙离子的荧光探针负载培养的小鼠皮质神经元标记物 ,应用激光共聚焦技术检测缺血条件和加用氟桂利嗪后细胞内荧光强度 (△FI)的变化。结果 给予缺血处理 10min后 ,神经元胞浆内钙离子△FI明显升高(17.5 2± 3.16 ) ;经氟桂利嗪预处理后 ,可以明显抑制钙离子的△FI(9.5 5± 2 .10 ) ,差异具有显著性意义 (P <0 .0 1) ,氟桂利嗪对无钙培养液缺血神经元胞浆内的钙离子浓度亦有抑制作用。结论 缺血状态下 ,神经元内钙离子明显升高 ,氟桂利嗪抑制胞浆内钙离子的升高 ,除通过阻断胞膜的电压敏感性钙通道外 ,可能还影响细胞内钙池的钙释放功能。

Objective To study the changes of intracellular free Ca 2+ and the effect of flunarizine on the calcium ion influx in cortical neurons under ischemia-like condition.Methods Mouse neurons were cultured and loaded with Flu-3/AM.I was measured by fluorescence intensity (△FI) in each cell by means of confocal laser scanning microscopy before and after the treatment with flunarizine.Results After 10min ischemia-like condition,a rapid increase in △FI was observed(17.52±3.16).The △FI in ischemic group was higher than that in flunarizine treatment group(9.55± 2.10),with significant difference(P<0.01). The △FI of therapy group neurons in Ca 2+ -free culture medium was significantly lower than that of ischemic group (P< 0.05). Conclusions There was a rapid increase in △FI in neurons under ischemia-like condition. Flunarizine reduced the increase inI in cortical neurons because it inhibited the calcium ion influx and might reduce release of Ca 2+ from calcium pools in the neurons.