氯胺酮对离体海马神经元生存及凋亡的影响

Effect on Ketamine of Activity and Apoptosis of Primary Hippocampal Neruons in Vitro

目的:探究不同浓度氯胺酮对离体原代海马神经元生存活力和凋亡的影响,并初步探讨其机制。方法:取原代培养7d的海马神经元随机分为6组,分别给予0μM、0.1μM、1μM、10μM、100μM、1 000μM不同浓度的氯胺酮干预。采用MTT法检测各组神经元的存活率,AnnexinV/PI双染色法检测凋亡率,Western blot分析凋亡相关基因Bcl-2、Bax蛋白表达量的变化。结果:与对照组相比,1μM、10μM和100μM组神经元存活率升高,凋亡率降低,Bcl-2蛋白的表达量升高,Bax蛋白表达量下降;而1 000μM组神经元存活率降低,凋亡率升高,Bcl-2蛋白表达降低,Bax蛋白表达升高。结论:氯胺酮对离体海马神经元的存活及凋亡的影响具有双向调节作用,低浓度的氯胺酮能够抑制神经元凋亡,提高细胞存活率;高浓度氯胺酮则促进神经元凋亡,降低存活率,可能是通过调节Bcl-2、Bax蛋白的表达起作用的。

Objective:To investigate the effects of different concentrations of ketamine on viability and apoptosis of the primary hippocampal neurons in vitro.Methods:The hippocampal neurons cultivated for 7 din vitro were divided into 6 groups and undergone different concentrations of ketamine:0μM,0.1μM,1μM,10μM,100μM and 1 000μM.Then,MTT assay was used to detect the viability of neurons,and the ratio of apoptosis was detected by AnnexinV/PI double staining.Western blot analysis was used to detect the expression levels of Bcl-2,Bax which was related to apoptosis.Results:Compared with control group,the viabilities of neurons were significantly increased in 1μM,10μM and 100μM ketamine groups and decreased in 1 000μM group(P<0.05);the neuronal apoptosis rates were decreased in 1μM,10μM and 100μM ketamine groups and increased in 1 000μM group(P<0.05);the expression of Bcl-2 were increased and Bax were decreased in 1μM,10μM and 100μM ketamine,and in 1 000μM group,the expression of Bcl-2 was lower and Bax was higher than that in control group.Conclusion:The effect of ketamine on viability and apoptosis of hippocampal neurons were bidirectional.Low concentrations of ketamine can inhibit the apoptosis and improve the viability of neurons,and high concentrations of ketamine induces apoptosis of hippocampal neurons through changing the expression of Bcl-2 and Bax.