小鼠皮质神经元类缺血/再灌注后氟桂利嗪对胞质内游离钙离子和神经元凋亡的实验研究

目的研究类缺血/再灌注后不同时间点神经元胞质内游离钙离子及神经元凋亡比例变化,探讨类缺血/再灌注后神经元的凋亡与神经内钙离子的关系及氟桂利嗪的治疗意义.方法利用Ca2 指示剂Flu-3/AM作为细胞内钙离子的荧光探针负载培养的神经元,共聚焦技术检测细胞内荧光强度的变化,原位末端标记法(TUNEL)观察神经元类缺血再灌注后不同时间点神经元凋亡情况.结果与类缺血/再灌注组相比,氟桂利嗪对类缺血/再灌注后神经元胞质内游离钙离子浓度增高和神经元凋亡有显著抑制作用(P＜0.05);再灌注3 h两组无显著差异.结论氟桂利嗪可明显抑制类缺血/再灌注后神经元胞质内钙离子的升高,减少神经元凋亡比例.

The intracellular Ca2+ and apoptosis in cortical neurons in ischemic/reperfusion-like conditions

Objective To study the changes of intracellular Ca 2 and apoptosis and protection of flunarizine on cortical neurons in ischemic/reperfusion-like conditions.Methods Neurons were cultured and loaded with Flu-3/AM.[Ca 2 ] I was measured by fluorescent intensity (FI) in each cell with confocal microscopy. Terminal deoxynucleotidyl transferase mediated nick end labeling (TUNEL) method was adopted to investigate to the apoptotic cells. Results There was obvious difference in intracellular Ca 2 and the number of apoptotic cells between ischemic/reperfusion-like group and flunarizine treatment group (P< 0.05), and at 3 h following ischemic/reperfusion-like, obvious difference was not observed between the two groups. Conclusions Flunarizine reduced the increase of [Ca 2 ] I and inhibited the apoptosis of ischemia/reperfusion-like neurons.