不同剂量拉莫三嗪对体外培养海马神经细胞存活及其BDNF和P-CREB表达的影响

目的:观察不同浓度拉莫三嗪(Lamotrigine,LTG)对体外培养海马神经细胞存活影响及脑源性神经生长因子(Brain derived neurotrophic factor,BDNF)和磷酸化cAMP反应元件结合(Phosphorylated cAMP response element binding protein,p-CREB)的表达变化.从细胞水平角度探索拉莫三嗪对认知功能的可能影响.方法:取培养8 d的成熟海马神经细胞,随机分为拉莫三嗪低剂量组(4mg/L)、拉莫三嗪中剂量组(8mg/L)、拉莫三嗪高剂量处理组(12mg/L)和空白对照组(0.1%DMSO).维持浓度培养4 d后,MTT法检测细胞存活数量,免疫组化检测神经细胞BDNF和p-CREB.结果:(1)不同浓度拉莫三嗪处理组与空白对照组之间比较,细胞存活率有明显差异(P<0.05).而3个不同剂量拉莫三嗪组之间的细胞存活率没有显著差异(P>0.05).(2)与正常对照组比12 mg/L拉莫三嗪培养神经细胞,p-CREB和BDNF的表达显著降低(P<0.05),而4、8 mg/L拉莫三嗪剂量组无差异(P>0.05).(3)加入拉莫三嗪12 mg/L培养神经细胞BDNF和p-CREB的表达明显低于4、8 mg/L拉莫三嗪剂量组(P<0.05).结论:在4～12 mg/L的剂量范围内,拉莫三嗪均可造成体外培养神经细胞死亡,高浓度拉莫三嗪可通过神经细胞内BDNF和p-CREB的表达变化,影响神经细胞存活.

Effect of lamotrigine of different concentration on the survival rate and the expression of BDNF and p-CREB in the cultured hippocampus neuron

Objective:To explore the possible effections on neuron survival rate and the change of BDNF and p-CREB induced by different concentrations of lamotrigine. Methods: The hippocampus was harvested from newborn Sprague - Dawley rats. Primary hippocampal neuron cultured for 8 days in vitro were used and divided randomly into four groups: low concentration (4 mg/L) group, middle concentration (12 mg/L) group, high concentration (12 mg/L) group and 0.1%DMSO as control. The BDNF and p-CREB levels were determined by immunohistochemistry and the cell survival rate was tested by MTT. Results: (1)compared with that in the control group, the survival of the neuron cell in all the lamotrigine groups decreased significantly (P<0.05),but the differences of survival rate was not great among the three lamotrigine groups. (2) Compared with the control group, expression of BDNF and p-CREB on the cell in the high concentration group were significantly different from those in the control group (P<0.05). (3) BDNF and p-CREB expressing on the cell in the 12 mg/L lamolrigine concentration group were significantly different from those in the 4 mg/L and 8 mg/L lamotrigine concentration group (P<0.05). Conclusion: In the leveT of 4-12 mg/L, lamtrigine can cause the neural cell death in vitro and the high concentration of lamotrigine (12 mg/L) causes the cell death by changing the expression of BDNF and p-CREB