酮体β-羟基丁酸减轻癫痫持续状态模型大鼠离体海马神经元的损伤

The mechanism of β-hydroxybutyric acid attenuating the damage of isolated hippocampal neurons of rats in status epileptic us model

目的探讨β-羟丁酸(BHB)对离体癫痫持续状态模型海马神经元的保护作用及可能机制。方法体外原代培养14 d神经元细胞,通过无镁灌流制成癫痫持续状态体外模型。加入不同浓度BHB(0 mmol/L、2 mmol/L、4 mmol/L、8 mmol/L)后采用MTT法测定神经元细胞的活性,通过流式细胞学方法测定加入BHB后神经细胞的凋亡情况及细胞中的活性氧簇表达情况。结果MTT实验示无镁模型体外神经元细胞活力明显下降,提示无镁灌流制成癫痫持续状态模型。高剂量组BHB处理后细胞活力明显改善,差异有统计学意义(P均<0.01)。流式细胞学方法示神经元在无镁环境下早期凋亡(P<0.05)、晚期凋亡(P<0.01)及凋亡总数(P<0.01)均增加。高剂量组早期凋亡及凋亡总数较模型组下降,差异有统计学意义(P均<0.05)。且活性氧簇水平下降,差异有统计学意义(P<0.05)。结论高浓度BHB能增加神经元活力,减少神经细胞凋亡,对癫痫持续状态体外培养海马神经元活性有保护作用,并可能通过影响活性氧簇途径起作用。

Objective To investigate the protective effect of ketone bodyβ-hydroxybutyrate(BHB)on hippocampal neurons of rats in status epilepticus(SE)model and its possible mechanism.Methods Neurons cultured in vitro for 14 days were given magnesium-free perfusion to make a SE model.The MTT was used to measure the viability of neurons after adding different concentrations of BHB.The apoptosis of neurons and the reactive oxygen species(ROS)were measured by flow cytometry.Results The neuronal viability in the SE model decreased significantly,suggesting that the magnesiumfree induced neuronal discharge leads to cell damage.The neuronal viability of the 8 mmol/L group treated with BHB was significantly improved compared with the 0 mmol/L,2 mmol/L,and 4 mmol/l BHB groups(all P<0.01).In vitro primary cultured neurons increased early apoptosis(P<0.05),late apoptosis(P<0.01),and the total number of apoptosis(P<0.01)in the magnesium-free groups.After adding 8 mmol/L BHB,the early apoptosis and the total number of apoptosis decreased compared with the group without BHB,and the difference was statistically significant(all P<0.05).The ROS level was significantly increased after magnesium-free perfusion(P<0.01).Compared with the BHB 0 mmol/L,the ROS level of the BHB 8 mmol/l group was decreased,and the difference was statistically significant(P<0.05).Conclusion High concentration of BHB can increase neuronal viability,reduce neuronal apoptosis,and protect the hippocampal neurons cultured in vitro in status epilepticus,and may act through ways affecting ROS.