白藜芦醇对难治性癫痫细胞模型神经元损伤及神经突起的影响

Effects of resveratrol on neuronal damage and neurite in cell model of intractable epilepsy

目的探讨白藜芦醇(Res)对难治性癫痫细胞模型的神经元损伤及神经突起形态学变化的影响。方法将体外培养至第10天的大鼠乳鼠海马神经元分为对照组、模型组、10μmol/L Res组、20μmol/L Res组、40μmol/L Res组及60μmol/L Res组。对照组采用正常细胞的细胞外液培养3 h后恢复维持培养液,模型组采用无镁细胞外液培养3 h后恢复维持培养液,4个Res组经无镁细胞外液培养3 h后加入相应浓度的Res培养液。检测建立模型后24 h各组细胞上清液乳酸脱氢酶(LDH)的活性,光学显微镜下观察各组神经元及神经突起的形态学变化。结果与对照组相比,模型组及各浓度Res组的LDH活性显著增加(P<0.05);与模型组相比,10μmol/L、20μmol/L、40μmol/L Res组LDH活性明显降低(P<0.05),其中以40μmol/L Res组最低;60μmol/L Res组与模型细胞的LDH活性比较,差异无统计学意义(P>0.05)。经建立模型后24 h,光学显微镜下可见模型组神经突起之间相互迁移聚集,突起连接成"网格样";40μmol/L Res组出现少部分神经突起迁移靠近聚集,大部分的突起连接分布均匀。结论 Res能减少难治性癫痫细胞模型的神经元损伤,抑制异常神经突起连接的形成。

Objective To explore the effects of resveratrol （Res） on the neuronal damage and morphologic change of neurite in cell model of intractable epilepsy. Methods On the 10th day of culture in vitro,the hippoeampal neurons of neonatal rats were divided into control group, model group, 10 μmol/L Res group, 20 μmoL/L Res group ,40μmoL/L Res group and 60μ mol/L Res group. Control group and model group continued to be cultured with feeding medium after 3 hours of culturing with normal extracellular fluid and magnesium-flee extraeellular fluid respectively. Res culture medium with corresponding concentrations was added to the four Res groups after 3 hours of culturing with magnesium-free extracellular fluid. After 24 hours of modeling, the lactic dehydrogenase （LDH） activity of cell supematant was detected in all groups, and the morphologic changes of neuron and neurite were observed in each group with optical microscope. Results Compared to control group ,model group and four Res groups had significantly increased activities of LDH（P 〈 0.05 ）. The activities of LDH in 10μmol/L Res group ,20 μmoL/L Res group and 40 μmoL/L Res group decreased significantly compared to that in the model group （ P 〈 0.05 ） , and the lowest activity was observed in 40 p, moL/L Res group. But there was no statistical difference in the activity of LDH between 60 μmol/L Res group and model group （P 〉 0.05 ）. After 24 hours of modeling, the observation under optical microscope showed that neurites migrated closely to each other and connected to be grid-shaped in the model group, but a few neurites migrated closely to each other and most of neurites connections uniformly distributed in 40μmol/L Res group. Conclusion Res can reduce the neuronal damage in the cell model of intractable epilepsy, and can inhibit the formation of abnormal neurite connections.